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.

Zoliflodacin: a hope to treat antibiotic-resistant Neisseria gonorrhoeae

Background:Neisseria gonorrhoeae is a gram-negative diplococcus that leads to sexually transmitted infection.N.gonorrhoeae is an obligate human pathogen that causes infection to the mucus-secreting epithelial cells both in males and females.In 2017 the center for disease control and the World Health Organization published the list of global priority pathogens-12 with denting therapeutic options,including antibiotic-resistant N.gonorrhoeae.Methods:we thoroughly characterized zoliflodacin antibiotic,its clinical trials and effect on human health by using different keywords like“zoliflodacin”,“COVID-19”,“clinical trials”from different data sources like Pub-Med,Google-Scholar,and Science-Direct.Results:Zoliflodacin shows a therapeutic approach against N.gonorrhoeae.It acts by inhibiting bacterial type 2 topoisomerase with the binding sites in bacterial gyrase.It shows promising results against N.gonorrhoeae.Zoliflodacin is effective in treating gonococcal urogenital and rectal infection.Conclusion:Currently,antibiotic is the only option to treat N.gonorrhoeae with no vaccine available to treat it.The new drug,zoliflodacin,specifically targets antibiotic-resistant gonorrhea and it has given a hope to researchers.This review elaborates the discovery of zoliflodacin,its mechanism of action,current clinical trials,and its effectiveness.

Determining antimicrobial resistance profiles and identifying novel mutations of Neisseria gonorrhoeae genomes obtained by multiplexed MinION sequencing

Gonorrhea is one of the most common sexually transmitted diseases worldwide. To cure infection and prevent transmission,timely and appropriate antimicrobial therapy is necessary. Unfortunately, Neisseria gonorrhoeae, the etiological agent of gonorrhea, has acquired nearly all known mechanisms of antimicrobial resistance(AMR), thereby compromising the efficacy of antimicrobial therapy. Treatment failure resulting from AMR has become a global public health concern. Whole-genome sequencing is an effective method to determine the AMR characteristics of N. gonorrhoeae. Compared with next-generation sequencing, the MinION sequencer(Oxford Nanopore Technologies(ONT)) has the advantages of long read length and portability. Based on a pilot study using MinION to sequence the genome of N. gonorrhoeae, we optimized the workflow of sequencing and data analysis in the current study. Here we sequenced nine isolates within one flow cell using a multiplexed sequencing strategy. After hybrid assembly with Illumina reads, nine integral circular chromosomes were obtained. By using the online tool Pathogenwatch and a BLAST-based workflow, we acquired complete AMR profiles related to seven classes of antibiotics. We also evaluated the performance of ONT-only assemblies. Most AMR determinants identified by ONT-only assemblies were the same as those identified by hybrid assemblies. Moreover, one of the nine assemblies indicated a potentially novel antimicrobial-related mutation located in mtrR which results in a frame-shift, premature stop codon, and truncated peptide.In addition, this is the first study using the MinION sequencer to obtain complete genome sequences of N. gonorrhoeae strains which are epidemic in China. This study shows that complete genome sequences and antimicrobial characteristics of N.gonorrhoeae can be obtained using the MinION sequencer in a simple and cost-effective manner, with hardly any knowledge of bioinformatics required. More importantly, this strategy provides us with a potential approach to discover new AMR determinants.

Using RAPD in Neisseria gonorrhoeae genotyping and transmission detection

The aim of this paper is to develop an applic-able Random Amplified Polymorphic DNAs(RAPD)method for genotyping Neisseria gonorrhoeae strain,and discuss the possibility of using the RAPD method to trace N.gonorrhoeae strain transmission route.Four different pretreatment methods were used on the N.gonorrhoeae genomic DNA component,and the best adaptive extract method was selected for RAPD.Different RAPD primers sequence was used for amplification and their differenti-ating capabilities for N.gonorrhoeae strains were com-pared.Applicable RAPD primer was selected for N.gonorrhoeae genotyping and then applied into transmis-sion detection.The results show that the so called cetyl-trimethylammonium bromide(CTAB)method for extracting genomic DNA could give integrated genomic DNA and give out relatively better RAPD fingerprint maps,subsequently,using selected RAPD primer could give out a group of amplification polymerase chain reac-tion bands.The fingerprint maps from different N.gonor-rhoeae strains were distinctive.Some main segments were common to all the N.gonorrhoeae strains tested.Some segments were different among the N.gonorrhoeae strains.According to the fingerprint maps and similarity index of different N.gonorrhoeae isolates,isolates from a pair of sex-partners were very similar.Based on these findings,the best extracting method and suitable RAPD primer were chosen.The RAPD fingerprint maps could type N.gonorrhoeae effectively and could be used as an additional approach in molecular epidemiology for tracing infection sources.

The relationship between sexually transmitted microorganisms and seminal quality in asymptomatic men

Objective:To detect DNA of different microorganisms,in semen samples from apparently healthy men and correlate their presence with seminal quality.Methods:Semensamples from 81 healthy volunteers were collected,and semen parameters were analyzed.DNA extraction was performed using the phenol-chloroform technique,and the micro-organisms were detected by the amplification of specific primers using polymerase chain reaction.Results:DNA from at least one of the microorganisms was detected in 78 samples.The most frequent microorganism found in semen were:Lactobacillus spp.(70%),Neisseria gonorrhoeae(N.gonorrhoeae)(36%),Streptococcus epidermidis(64%),Klebsiella pneumoniae(56%),Staphylococcus aureus(32%),Chlamydia trachomatis(C.trachomatis)(28%),Pseudomonas aeru-ginosa(27%).The seminal parameters of all semen samples were over the lower reference values fornormal semenanalysis.To compare with negative samples,seminal volume was higher for the Escherichia coli positive samples and lower for Pseudomonas aeruginosa positive samples.Semen samples positive for Staphylococcus aureus had worse sperm morphology.The frequency of progressive motility was higher in positive samples for N.gonorrhoeae and C.trachomatis.Positive semen samples for C.trachomatis had a higher concentration per milliliter.Conclusion:It is common to find microorganisms in semen of asymptomatic men,including those responsible for sexually transmitted infections.Antimicrobial treatment is recommended only in those individuals with a sexually transmitted infection(C.trachomatis and N.gonorrhoeae)and always promote condom use.

Surveillance of Gonococcal Infections with Assessment of Their Antibiotic Susceptibility Pattern

Context: Gonococcal infections are prevalent throughout the world and remain a serious public health problem, with an estimated 106.1 million new infections per year. Over recent years, the gonococcus has rapidly acquired resistance to commonly used antibiotics and very few treatment options remain;hence, regular monitoring of antimicrobial resistance is very important. Methods and Material: The study was conducted from January 2012 to March 2013 after approved by IRB & HREC in patients attending O&G and Skin & VD with the complaints of vaginal discharge, cervical discharge, urethral discharge, dyspareunia, burning micturition, lower abdominal pain & infertility. Endocervical swabs and urethral swabs were collected and cultured on chocolate agar andmodified Thayer martin medium identification of organism was done by biochemical tests and antimicrobial susceptibility testing was done by Kirby-Bauer disk diffusion method as per CLSI guidelines. Results: FiveNeisseria were isolated in male (2.3%) patients of reproductive age group (20 - 30years) with one found reactive for HIV. All patients had history of heterosexual contact of 10 - 12 days before onset of urethral discharge and burning micturition. All isolates were resistant to Penicillin, Tetracycline and Ciprofloxacin. Only one isolate was resistance to Azithromycin. Conclusions: There is a development of resistance in gonococcal isolates to third generation cephalosporin and azithromycin and these are the drugs that are given for syndromic management of cases as per NACO guidelines. So surveillance for antimicrobial resistance is crucial for monitoring the emergence and spread of antibiotic resistance in gonococcal isolates.

Global Transmission of the penA Allele 60.001–Containing High-Level Ceftriaxone-Resistant Gonococcal FC428 Clone and Antimicrobial Therapy of Associated Cases:A Review

Neisseria gonorrhoeae is a multidrug-resistant bacterial pathogen for which ceftriaxone is the only remaining recommended first-line therapy.However,ceftriaxone susceptibility has been waning in a number of countries over the last decade and ceftriaxone treatment failures have been reported,commonly as a result of sporadic high-level ceftriaxone-resistant strains.In recent years,N.gonorrhoeae strains associated with the high-level ceftriaxone-resistant FC428 clone or strains that acquired its main ceftriaxone resistance determinant,penA allele 60.001,have shown global transmission,resulting in ceftriaxone treatment failure in a number of cases.The FC428 clone was first encountered in Japan in 2015 and subsequently in China,Europe,Australia,North America and Southeast Asia afterward.Strains associated with the FC428 clone commonly display a ceftriaxone minimum inhibitory concentration of 0.5-1 mg/L.However,where penA alleles encountered in sporadic high-level ceftriaxone-resistant isolates induce an in vitro growth defect,penA allele 60.001 does not seem to affect in vitro growth.The limited impact of penA allele 60.001 on biological fitness might be associated with its successful global transmission.Although the FC428 clone displays high-level ceftriaxone resistance,most gonorrhea cases associatedwith this clone were still successfully curedwith ceftriaxonewhen intramuscular or intravenous doses of 500mg to 2 g were used.A successful alternative therapy seems to be ertapenem given at 1-g doses,although further clinical studies are required to validate ertapenemefficacy.This review summarizes the global transmission of strains associated with the FC428 clone and antimicrobial treatment of associated cases.

Determination of Fusidic Acid Susceptibility of Multidrug-resistant Gonococcal Isolates from China

The currently recommended gonococcal therapies are rapidly losing efficacy and;therefore,novel options for treatment are urgently required to ensure future treatment remains available.Here we investigated the susceptibility of 379 contemporary clinical Neisseria gonorrhoeae isolates from China for susceptibility to fusidic acid.These isolates include numerous multidrug-resistant strains with resistance or reduced susceptibility against ceftriaxone and/or azithromycin,antimicrobials that are currently recommended as a first-line dual therapy.Although official susceptibility breakpoints have not been established,all gonococcal isolates appeared to be susceptible to fusidic acid,with the majority of isolates displaying a minimal inhibitory concentration(MIC)of 0.25 or 0.5mg/L.Overall,the MIC90 was 0.5mg/L,with a MIC range of 0.08-2mg/L.Importantly,no correlation between fusidic acid susceptibility and susceptibility to ceftriaxone or azithromycin was observed.Further analysis of gonococcal survival curves indicated that fusidic acid was only mildly bactericidal at the highest tested concentrations(4
MIC).In conclusion,fusidic acid displayed consistent antimicrobial activity against N.gonorrhoeae and,given the absence of cross-resistance with ceftriaxone and azithromycin,might be interesting for further evaluation of single or dual antimicrobial therapies in combination with ceftriaxone or azithromycin.

Antibiotic Resistance and Treatment Options for Multidrug-Resistant Gonorrhea

Gonorrhea is a sexually transmitted disease with a high global incidence.Its causative agent,Neisseria gonorrhoeae,has shown a remarkable flexibility to adapt and become resistant to all antimicrobials introduced over the past century for gonococcal therapy.The currently last available first-line therapy that is recommended in most countries is ceftriaxone.However,resistance levels against ceftriaxone are rising globally and incidences of confirmed treatment failure are increasingly encountered,particularly with the global spreading of the ceftriaxone-resistant FC428 clone in recent years.Resistance against most antimicrobials has been the result of adaptive genomic mutations that reduce affinity of the antimicrobial to its target protein or rRNA,although most multidrug-resistant strains also appear to have an upregulated multidrug efflux pump.To ensure gonococcal treatment remains available in the future,alternative therapies are urgently needed.Therefore,both alternative clinically approved antimicrobials and novel antimicrobials have been intensely studied both in gonococcal susceptibility analyses and clinical efficacy trials.Although there have been some limited successes,all studied alternative therapies that reached clinical trials have displayed some shortcomings in their efficacy against pharyngeal infections and/or overlapping resistance determinants with previously or currently used antimicrobials.This review summarizes the development of gonococcal antimicrobial resistance over the past century,describes the mechanisms involved in antimicrobial resistance,and provides an overview of the alternative therapies that have been under investigation this past decade.