Neisseria Species Research Articles

A role for fibroblast growth factor receptor 1 in the pathogenesis of Neisseria meningitidis

Neisseria meningitidis (the meningococcus) remains an important cause of human disease, including meningitis and sepsis. Adaptation to the host environment includes many interactions with specific cell surface receptors, resulting in intracellular signalling and cytoskeletal rearrangements that contribute to pathogenesis. Here, we assessed the interactions between meningococci and Fibroblast Growth Factor Receptor 1-IIIc (FGFR1-IIIc): a receptor specific to endothelial cells of the microvasculature, including that of the blood-brain barrier. We show that the meningococcus recruits FGFR1-IIIc onto the surface of human blood microvascular endothelial cells (HBMECs). Furthermore, we demonstrate that expression of FGFR1-IIIc is required for optimal invasion of HBMECs by meningococci. We show that the ability of N. meningitidis to interact with the ligand-binding domain of FGFR1-IIIc is shared with the other pathogenic Neisseria species, N. gonorrhoeae, but not with commensal bacteria including non-pathogenic Neisseria species.

2020 FASEB Science Research Conference on Microbial Glycobiology, July 13-14, 2020.

2020 FASEB Science Research Conference on Microbial Glycobiology, July 13-14, 2020.

Evaluation of the SpeeDx ResistancePlus® GC and SpeeDx GC 23S 2611 (beta) molecular assays for prediction of antimicrobial resistance/susceptibility to ciprofloxacin and azithromycin in Neisseria gonorrhoeae.

Accurate molecular assays for prediction of antimicrobial resistance (AMR)/susceptibility in Neisseria gonorrhoeae (Ng) can offer individualized treatment of gonorrhoea and enhanced AMR surveillance. We evaluated the new ResistancePlus® GC assay and the GC 23S 2611 (beta) assay (SpeeDx), for prediction of resistance/susceptibility to ciprofloxacin and azithromycin, respectively. Nine hundred and sixty-seven whole-genome-sequenced Ng isolates from 20 European countries, 143 Ng-positive (37 with paired Ng isolates) and 167 Ng-negative clinical Aptima Combo 2 (AC2) samples, and 143 non-gonococcal Neisseria isolates and closely related species were examined with both SpeeDx assays. The sensitivity and specificity of the ResistancePlus® GC assay to detect Ng in AC2 samples were 98.6% and 100%, respectively. ResistancePlus® GC showed 100% sensitivity and specificity for GyrA S91 WT/S91F detection and 99.8% sensitivity and specificity in predicting phenotypic ciprofloxacin resistance. The sensitivity and specificity of the GC 23S 2611 (beta) assay for Ng detection in AC2 samples were 95.8% and 100%, respectively. GC 23S 2611 (beta) showed 100% sensitivity and 99.9% specificity for 23S rRNA C2611 WT/C2611T detection and 64.3% sensitivity and 99.9% specificity for predicting phenotypic azithromycin resistance. Cross-reactions with non-gonococcal Neisseria species were observed with both assays, but the analysis software solved most cross-reactions. The new SpeeDx ResistancePlus® GC assay performed well in the detection of Ng and AMR determinants, especially in urogenital samples. The GC 23S 2611 (beta) assay performed relatively well, but its sensitivity, especially for predicting phenotypic azithromycin resistance, was suboptimal and further optimizations are required, including detection of additional macrolide resistance determinant(s).

Allelic polymorphisms in a glycosyltransferase gene shape glycan repertoire in the O-linked protein glycosylation system of Neisseria.

Glycosylation of multiple proteins via O-linkage is well documented in bacterial species of Neisseria of import to human disease. Recent studies of protein glycosylation (pgl) gene distribution established that related protein glycosylation systems occur throughout the genus including nonpathogenic species. However, there are inconsistencies between pgl gene status and observed glycan structures. One of these relates to the widespread distribution of pglG, encoding a glycosyltransferase that in Neisseria elongata subsp. glycolytica is responsible for the addition of di-N-acetyl glucuronic acid at the third position of a tetrasaccharide. Despite pglG residing in strains of N. gonorrhoeae, N. meningitidis and N. lactamica, no glycan structures have been correlated with its presence in these backgrounds. Moreover, PglG function in N. elongata subsp. glycolytica minimally requires UDP-glucuronic acid (GlcNAcA), and yet N. gonorrhoeae, N. meningitidis and N. lactamica lack pglJ, the gene whose product is essential for UDP-GlcNAcA synthesis. We examined the functionality of pglG alleles from species spanning the Neisseria genus by genetic complementation in N. elongata subsp. glycolytica. The results indicate that select pglG alleles from N. meningitidis and N. lactamica are associated with incorporation of an N-acetyl-hexosamine at the third position and reveal the potential for an expanded glycan repertoire in those species. Similar experiments using pglG from N. gonorrhoeae failed to find any evidence of function suggesting that those alleles are missense pseudogenes. Taken together, the results are emblematic of how allelic polymorphisms can shape bacterial glycosyltransferase function and demonstrate that such alterations may be constrained to distinct phylogenetic lineages.

Abstract 1163: Lung and salivary microbiome in electronic cigarette users, never-smokers, and smokers: A pilot cross-sectional study

Abstract Background: Little is known about the microbiomes of the lung and oral cavity with electronic cigarette (e-cig) use and how they compare to those of smokers and never-smokers. E-cigs have been promoted as a safe alternative to smoking cigarettes. Given the recent outbreak of E-cigarette or Vaping product use Associated Lung Injury (EVALI) there is an urgent need for understanding the biological effects of e-cig use on the lung and oral cavity, including effects on the microbiome. Previous studies have been limited to 16S-rRNA sequencing which were used to detect bacteria genera. In this study, we used metatranscriptome profiling to study differentially abundant bacteria species in the oral and lung microbiome of never-smokers, smokers, and e-cig users. Methods: A cross-sectional study of 10 never-smokers, 8 cigarette smokers, and 10 e-cig users was conducted, with saliva and bronchoalveolar lavage (BAL) collected for each study participant. RNA was extracted from saliva and BAL samples for total transcriptome RNA-seq analysis. Sequences were aligned with bowtie2 v.2.2.8 to the human genome (hg19) and non-aligned reads were aligned and annotated using NCBI metagenomes database and Kraken v.1. Differences in the microbiome by smoking status were determined by pairwise comparisons using limma-voom with FDR q-value cutoffs <0.2. Results: The distribution of richness and evenness of bacterial communities measured by Shannon diversity in our metatranscriptome data did not significantly differ between the three smoking status groups. When comparing levels of bacteria species between groups in the saliva, 234 were differentially abundant between smokers and never-smokers, and 39 were differentially abundant between smokers and e-cig users. In the lung, 87 bacterial species were differentially abundant between smokers and never-smokers and 36 were differentially abundant between smokers and e-cig users. Notably, no bacteria species were differentially abundant when comparing e-cig users and never-smokers in both the saliva and lung samples. There are 50 bacterial species found to be differentially abundant in both the lung and saliva samples, 47 of which are decreased in smokers. These 47 bacteria species included common commensal oral microbiome species such as Haemophilus parainfluenzae, Capnocytophaga gingivalis and Neisseria species. The 3 species that were increased in smokers were Lactobacillus species. Conclusion: Our findings suggests that smoking cigarettes may alter populations of common commensal species in both the oral and lung microbiome. The lack of differentially abundant bacterial species between electronic cigarette users and never-smokers indicates that e-cigs may alter bacterial species to a lesser extent than smoking. Citation Format: Kevin L. Ying, Min-Ae Song, Daniel Y. Weng, Quentin A. Nickerson, Joseph P. McElory, Theodore M. Brasky, Noah B. Whiteman, Mark D. Wewers, Jo L. Freudenheim, Ewy A. Mathé, Peter G. Shields. Lung and salivary microbiome in electronic cigarette users, never-smokers, and smokers: A pilot cross-sectional study [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1163.

Characteristics of Neisseria Species Colonized in the Human’s Nasopharynx

Context: Neisseria meningitidis is the causative agent of a life-threatening infection with high mortality and morbidity worldwide. The most common types of this bacterium are serogroups A, B, C, W135, X, and Y. Although in some countries, such as Iran, the meningococcal meningitis has been well monitored and controlled by the use of divalent and quadrivalent vaccines, other fatal infections caused by these bacteria are still an important threat. For the above reason, this review focused on the differences of Neisseria characteristics, particularly in capsular composition, pathogenic and commensal stages to a better understanding of how to manage Neisseria infections. Evidence Acquisition: In this review, PubMed, EMBASE, ScienceDirect, Scopus, and Google Scholar were searched for English-language publications on pathogenic or commensal strains of Neisseria, meningococcal disease, Neisseria biology, genetic diversity, molecular typing, serogroups, diagnostic, and epidemiology around the world up to July 2019. All articles and academic reports in the defined area of this research were considered too. The data were extracted and descriptively discussed. Results: We included 85 studies in the survey. The data analysis revealed that the distribution of meningococcal serogroups was different regionally. For example, the serogroups C and W-135 accounted for Africa and Latin America regions, serogroup B in the European countries, and rarely in the Western Pacific, and serogroups A and C were dominant in Asian countries. Although data set for laboratory-based diagnosis of N. meningitidis are available for all countries, only 30% of the countries rely on reference laboratories for serogroup determination, and more than half of the countries lack the ability of surveillance system. Nevertheless, molecular detection procedure is also available for all countries. The use of the meningococcal vaccine is a variable country by country, but most countries have applied the meningococcal vaccine, either divalent or quadrivalent, for the protection of high-risk groups. Conclusions: Owing to the geographical distribution of N. meningitidis serogroups in circulating, each country has to monitor for changes in serogroups diversity and its control management. Furthermore, laboratories should scale up the epidemiology and disease burden. It should be mentioned that quadrivalent meningococcal vaccines reduce the meningococcal disease burden sharply.

Abstract A34: Lung and salivary microbiome in electronic cigarette users, never-smokers, and smokers: A pilot cross-sectional study

Abstract Background: Little is known about the microbiomes of the lung and oral cavity with electronic cigarette (e-cig) use and how they compare to those of smokers and never-smokers. E-cigs have been promoted as a safe alternative to smoking cigarettes. Given the recent outbreak of E-cigarette or Vaping product use Associated Lung Injury (EVALI), there is an urgent need for understanding the biologic effects of e-cig use on the lung and oral cavity, including effects on the microbiome. Previous studies have been limited to 16S-rRNA sequencing, which was used to detect bacteria genera. In this study, we used metatranscriptome profiling to study differentially abundant bacteria species in the oral and lung microbiome of never-smokers, smokers, and e-cig users. Methods: A cross-sectional study of 10 never-smokers, 8 cigarette smokers, and 10 e-cig users was conducted, with saliva and bronchoalveolar lavage (BAL) collected for each study participant. RNA was extracted from saliva and BAL samples for total transcriptome RNA-seq analysis. Sequences were aligned with bowtie2 v.2.2.8 to the human genome (hg19) and nonaligned reads were aligned and annotated using NCBI metagenomes database and Kraken v.1. Differences in the microbiome by smoking status were determined by pairwise comparisons using limma-voom with FDR q-value cutoffs <0.2. Results: The distribution of richness and evenness of bacterial communities measured by Shannon diversity in our metatranscriptome data did not significantly differ between the three smoking status groups. When comparing levels of bacteria species between groups in the saliva, 234 were differentially abundant between smokers and never-smokers, and 39 were differentially abundant between smokers and e-cig users. In the lung, 87 bacterial species were differentially abundant between smokers and never-smokers and 36 were differentially abundant between smokers and e-cig users. Notably, no bacteria species were differentially abundant when comparing e-cig users and never-smokers in both the saliva and lung samples. There are 50 bacterial species found to be differentially abundant in both the lung and saliva samples, 47 of which are decreased in smokers. These 47 bacteria species included common commensal oral microbiome species such as Haemophilus parainfluenzae, Capnocytophaga gingivalis, and Neisseria species. The 3 species that were increased in smokers were Lactobacillus species. Conclusion: Our findings suggests that smoking cigarettes may alter populations of common commensal species in both the oral and lung microbiome. The lack of differentially abundant bacterial species between electronic cigarette users and never-smokers indicates that e-cigs may alter bacterial species to a lesser extent than smoking. Citation Format: Kevin L. Ying, Min-Ae Song, Daniel Y. Weng, Quentin A. Nickerson, Joseph P. McElroy, Theodore M. Brasky, Noah B. Whiteman, Mark D. Wewers, Jo L. Freudenheim, Ewy A. Mathe, Peter G. Shields. Lung and salivary microbiome in electronic cigarette users, never-smokers, and smokers: A pilot cross-sectional study [abstract]. In: Proceedings of the AACR Special Conference on the Microbiome, Viruses, and Cancer; 2020 Feb 21-24; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2020;80(8 Suppl):Abstract nr A34.

Evolutionary analysis of gyrA gene from Neisseria meningitidis bacterial strains of clonal complex 4821 collected in China between 1978 and 2016

BackgroundNeisseria meningitidis (N.meningitidis) bacteria belonging to clonal complex 4821 (CC4821) have been mainly reported in China and have been characterized by a high resistance rate to ciprofloxacin (CIP). The aim of this study was to assess the evolution of the DNA gyrase A (gyrA) gene from N.meningitidis CC4821 strains collected in China between 1978 and 2016. The complete sequence of gyrA gene from 77 strains are reported in this study and analyzed in the context of publicly available sequences from N. meningitidis of other CCs as well as other Neisseria species.ResultsThe phylogenetic analysis of CC4821 gyrA gene reveals at least 5 distinct genetic clusters. These clusters are not CC4821-specific showing that gyrA evolution is independent of CC4821 evolution. Some clusters contain sequences from other Neisseria species. Recombination within N.meningitidis strains and between Neisseria species was identified in SimPlot analysis. Finally, amino acid substitutions within GyrA protein were analyzed. Only one position, 91 (83 in E.coli gyrA gene), was linked to CIP resistance. Thirty-one additional putative resistance markers were identified, as amino acid substitutions were only found in resistant strains.ConclusionsThe evolution of gyrA gene of CC4821 N.meningitidis strains is not dependent on CC4821 evolution or on CIP resistance phenotype. Only amino acid 91 is linked to CIP resistance phenotype. Finally, recombination inter- and intra-species is likely to result in the acquisition of various resistance markers, 31 of them being putatively mapped in the present study. Analyzing the evolution of gyrA gene within CC4821 strains is critical to monitor the CIP resistance phenotype and the acquisition of new resistance markers. Such studies are necessary for the control of the meningococcal disease and the development of new drugs targeting DNA gyrase.

A diagnostic challenge in clinical laboratory: Misidentification of Neisseria subflava as Neisseria meningitidis by MALDI-TOF MS.

MALDI-TOF MS provides fast, easy to perform and cost-effective diagnosis in clinical microbiology laboratories, however in some cases results of MALDI-TOF MS should be confirmed with additional tests. This confirmation is especially important for causes of life-threatening infections like Neisseria meningitidis. In our laboratory, three isolates were identified as N. meningitidis by Bruker MALDI Biotyper (BD, USA) between April 2018 and March 2019 from clinical specimens of blood, sputum, and urine. 16S rRNA sequencing was performed for further investigation. Two of the isolates were identified as Neisseria subflava and only one was confirmed as N. meningitidis by sequencing. These results show that MALDI-TOF MS is not always reliable in the diagnosis of N. meningitidis and clinical microbiologists should confirm these results with additional tests. Also, clinical correlations should be determined. Accurate identification of this microorganism is very important because of the necessity of prophylactic antimicrobial usage and biosafety precautions. Enlarged databases of Neisseria species are needed to overcome this problem.

Commensal Neisseria cinerea impairs Neisseria meningitidis microcolony development and reduces pathogen colonisation of epithelial cells.

It is increasingly being recognised that the interplay between commensal and pathogenic bacteria can dictate the outcome of infection. Consequently, there is a need to understand how commensals interact with their human host and influence pathogen behaviour at epithelial surfaces. Neisseria meningitidis, a leading cause of sepsis and meningitis, exclusively colonises the human nasopharynx and shares this niche with several other Neisseria species, including the commensal Neisseria cinerea. Here, we demonstrate that during adhesion to human epithelial cells N. cinerea co-localises with molecules that are also recruited by the meningococcus, and show that, similar to N. meningitidis, N. cinerea forms dynamic microcolonies on the cell surface in a Type four pilus (Tfp) dependent manner. Finally, we demonstrate that N. cinerea colocalises with N. meningitidis on the epithelial cell surface, limits the size and motility of meningococcal microcolonies, and impairs the effective colonisation of epithelial cells by the pathogen. Our data establish that commensal Neisseria can mimic and affect the behaviour of a pathogen on epithelial cell surfaces.

Defective lytic transglycosylase disrupts cell morphogenesis by hindering cell wall de-O-acetylation in Neisseria meningitidis.

Lytic transglycosylases (LT) are enzymes involved in peptidoglycan (PG) remodeling. However, their contribution to cell-wall-modifying complexes and their potential as antimicrobial drug targets remains unclear. Here, we determined a high-resolution structure of the LT, an outer membrane lipoprotein from Neisseria species with a disordered active site helix (alpha helix 30). We show that deletion of the conserved alpha-helix 30 interferes with the integrity of the cell wall, disrupts cell division, cell separation, and impairs the fitness of the human pathogen Neisseria meningitidis during infection. Additionally, deletion of alpha-helix 30 results in hyperacetylated PG, suggesting this LtgA variant affects the function of the PG de-O-acetylase (Ape 1). Our study revealed that Ape 1 requires LtgA for optimal function, demonstrating that LTs can modulate the activity of their protein-binding partner. We show that targeting specific domains in LTs can be lethal, which opens the possibility that LTs are useful drug-targets.

Meningococcal Disease-Associated Prophage-Like Elements Are Present in Neisseria gonorrhoeae and Some Commensal Neisseria Species.

Neisseria spp. possess four genogroups of filamentous prophages, termed Nf1 to 4. A filamentous bacteriophage from the Nf1 genogroup termed meningococcal disease-associated phage (MDA φ) is associated with clonal complexes of Neisseria meningitidis that cause invasive meningococcal disease. Recently, we recovered an isolate of Neisseria gonorrhoeae (ExNg63) from a rare case of gonococcal meningitis, and found that it possessed a region with 90% similarity to Nf1 prophages, specifically, the meningococcal MDA φ. This led to the hypothesis that the Nf1 prophage may be more widely distributed amongst the genus Neisseria. An analysis of 92 reference genomes revealed the presence of intact Nf1 prophages in the commensal species, Neisseria lactamica and Neisseria cinerea in addition to the pathogen N. gonorrhoeae. In N. gonorrhoeae, Nf1 prophages had a restricted distribution but were present in all representatives of MLST ST1918. Of the 160 phage integration sites identified, only one common insertion site was found between one isolate of N. gonorrhoeae and N. meningitidis. There was an absence of any obvious conservation of the receptor for prophage entry, PilE, suggesting that the phage may have been obtained by natural transformation. An examination of the restriction modification systems and mutated mismatch repair systems with prophage presence suggested that there was no obvious preference for these hosts. A timed phylogeny inferred that N. meningitidis was the donor of the Nf1 prophages in N. lactamica and N. gonorrhoeae. Further work is required to determine whether Nf1 prophages are active and can act as accessory colonization factors in these species.

Mosaic structure of the penA gene in the oropharynx of men who have sex with men negative for gonorrhoea

The oropharynx represents a crucial site for the emergence of multi-drug resistance in Neisseria gonorrhoeae. The mosaic penA alleles, associated with decreased susceptibility to cephalosporins, have emerged by DNA recombination with partial penA genes, particularly those from commensal pharyngeal Neisseria species. Here, we investigated the prevalence of the mosaic structure of the penA gene in the oropharynx of men who have sex with men testing negative for pharyngeal gonorrhoea. From January 2016 to June 2018, 351 gonorrhoea-negative men who have sex with men attending a sexually transmitted infection clinic in Italy were enrolled. Pharyngeal swabs underwent a real-time polymerase chain reaction (PCR) for the detection of the mosaic penA gene. In case of positivity, PCR products were sequenced and searched against several sequences of Neisseria strains. Overall, 31 patients (8.8%) were found positive for the presence of the mosaic penA gene. The positivity was significantly associated with previous cases of pharyngeal gonorrhoea (relative risk [RR]: 3.56, 95% confidence interval 1.44–8.80) and with recent exposure to beta-lactams (RR: 4.29, 95% confidence interval 2.20–8.38). All penA-positive samples showed a high relatedness (90–99%) with mosaic-positive Neisseria strains. Our data underline that commensal Neisseria species of the oropharynx may be a significant reservoir for genetic material conferring antimicrobial resistance in N. gonorrhoeae.

Meningococcal Quinolone Resistance Originated from Several Commensal Neisseria Species.

Quinolone resistance is increasing in Neisseria meningitidis, with its prevalence in China being high (>70%), but its origin remains unknown. The aim of this study was to investigate the donors of mutation-harboring gyrA alleles in N. meningitidis A total of 198 N. meningitidis isolates and 293 commensal Neisseria isolates were collected between 2005 and 2018 in Shanghai, China. The MICs of ciprofloxacin were determined using the agar dilution method. The resistance-associated genes gyrA and parC were sequenced for all isolates, while a few isolates were sequenced on the Illumina platform. The prevalences of quinolone resistance in the N. meningitidis and commensal Neisseria isolates were 67.7% (134/198) and 99.3% (291/293), respectively. All 134 quinolone-resistant N. meningitidis isolates possessed mutations in T91 (n = 123) and/or D95 (n = 12) of GyrA, with 7 isolates also harboring ParC mutations and exhibiting higher MICs. Phylogenetic analysis of the gyrA sequence identified six clusters. Among the 71 mutation-harboring gyrA alleles found in 221 N. meningitidis isolates and genomes (n = 221), 12 alleles (n = 103, 46.6%) were included in the N. meningitidis cluster, while 20 alleles (n = 56) were included in the N. lactamica cluster, 27 alleles (n = 49) were included in the N. cinerea cluster, and 9 alleles (n = 10) were included in the N. subflava cluster. Genomic analyses identified the exact N. lactamica donors of seven mutation-harboring gyrA alleles (gyrA92, gyrA97, gyrA98, gyrA114, gyrA116, gyrA151, and gyrA230) and the N. subflava donor isolate of gyrA171, with the sizes of the recombinant fragments ranging from 634 to 7,499 bp. Transformation of gyrA fragments from these donor strains into a meningococcal isolate increased its ciprofloxacin MIC from 0.004 μg/ml to 0.125 or 0.19 μg/ml and to 0.5 μg/ml with further transformation of an additional ParC mutation. Over half of the quinolone-resistant N. meningitidis isolates acquired resistance by horizontal gene transfer from three commensal Neisseria species. Quinolone resistance in N. meningitidis increases in a stepwise manner.

Pharyngeal microbiome alterations during Neisseria gonorrhoeae infection.

Pharyngeal gonorrhoea is a common sexually transmitted infection among ‘men having sex with other men’ (MSM). Neisseria gonorrhoeae (NG) pharyngeal infections are usually characterized by the absence of symptoms, acting as an important reservoir for their further spread. To the best of our knowledge, no information about the composition of the pharyngeal microbiome during an ongoing NG infection is currently available. Therefore, in this study, we characterized the pharyngeal bacterial community profiles associated with NG infection in a well-selected cohort of HIV-negative MSM reporting unsafe oral intercourse. A total of 70 pharyngeal swabs were considered, comparing non-infected subjects (n = 45) versus patients with pharyngeal gonorrhoea (n = 25) whose microbiota composition was analyzed from pharyngeal swabs through sequencing of hypervariable V3-V4 regions of the 16S rRNA gene. The pharyngeal microbiome of all subjects was dominated by Prevotellaceae, Veillonellaceae and Streptococcaceae families. Patients with pharyngeal gonorrhoea harboured a pharyngeal microbiome quite similar to negative subjects. Nevertheless, when looking to less-represented bacterial species (relative abundance approximately 1% or less), an imbalance between aerobe and anaerobe microorganisms was observed in NG-infected patients. In particular, the pharyngeal microbiome of NG-positive individuals was richer in several anaerobes (e.g. Treponema, Parvimonas, Peptococcus, Catonella, Filifactor) and poorer in various aerobe genera (i.e. Pseudomonas, Escherichia), compared to non-infected controls. No significant differences were noticed in the distribution of commensal Neisseria species of the oropharynx between NG-positive and negative subjects. Metabolic variations induced by changes in the microbiome abundance were assessed by a functional prediction of the bacterial metabolic pathways: a more abundant involvement of D-glutamine and D-glutamate metabolism, carbohydrate metabolism, as well as a greater activation of the energy metabolism was observed in patients with pharyngeal gonorrhoea compared to non-infected individuals. Information about the bacterial composition of the pharyngeal microbiome in case of gonorrhoea could shed light on the pathogenesis of the infection and open new perspectives for the prevention and control of this condition.

Structural Basis for Evasion of Nutritional Immunity by the Pathogenic Neisseriae.

The pathogenic Neisseria species are human-adapted pathogens that cause quite distinct diseases. Neisseria gonorrhoeae causes the common sexually transmitted infection gonorrhea, while Neisseria meningitidis causes a potentially lethal form of bacterial meningitis. During infection, both pathogens deploy a number of virulence factors in order to thrive in the host. The focus of this review is on the outer membrane transport systems that enable the Neisseriae to utilize host-specific nutrients, including metal-binding proteins such as transferrin and calprotectin. Because acquisition of these critical metals is essential for growth and survival, understanding the structures of receptor-ligand complexes may be an important step in developing preventative or therapeutic strategies focused on thwarting these pathogens. Much can also be learned by comparing structures with antigenic diversity among the transporter sequences, as conserved functional domains in these essential transporters could represent the pathogens’ “Achilles heel.” Toward this goal, we present known or modeled structures for the transport systems produced by the pathogenic Neisseria species, overlapped with sequence diversity derived by comparing hundreds of neisserial protein sequences. Given the concerning increase in N. gonorrhoeae incidence and antibiotic resistance, these outer membrane transport systems appear to be excellent targets for new therapies and preventative vaccines.

Emergence of ceftriaxone-resistant Neisseria gonorrhoeae strains harbouring a novel mosaic penA gene in China.

The continuous emergence of ceftriaxone-resistant Neisseria gonorrhoeae strains threatens the effectiveness of current treatment regimens for gonorrhoea. The objective of the present study was to characterize three ceftriaxone-resistant N. gonorrhoeae strains with a novel mosaic penA allele isolated in China. Three ceftriaxone-resistant Neisseria gonorrhoeae strains (GC150, GC161 and GC208) isolated in 2017 were characterized by N. gonorrhoeae multiantigen sequence typing (NG-MAST), MLST and N. gonorrhoeae sequence typing for antimicrobial resistance (NG-STAR). Recombination analyses were performed using the SimPlot software. Three strains had the same antibiotic resistance profiles, with resistance to ceftriaxone (MIC 0.5 mg/L), ciprofloxacin (MIC 8.0 mg/L), penicillin (MIC 2.0 mg/L) and tetracycline (MIC 2.0-8.0 mg/L). STs were assigned as MLST7360, NG-MAST14292 and NG-STAR1611/NG-STAR1612. The penA gene of these three strains differed from previous ceftriaxone-resistant gonococcal strains and harboured a novel mosaic allele (penA-121.001). Like N. gonorrhoeae FC428, a widely disseminated ceftriaxone-resistant strain that was initially described in Japan in 2015, all strains also possessed substitutions A311V and T483S in PBP2, which are associated with resistance to ceftriaxone. Potential recombination events were detected in penA between N. gonorrhoeae strain FC428 and commensal Neisseria species. Our results provide further evidence that the commensal Neisseria species (Neisseria cinerea and Neisseria perflava) can serve as a reservoir of ceftriaxone resistance-mediating penA sequences in clinical gonococcal strains. The emergence of such strains may be the result of the interspecies recombination of penA genes between N. gonorrhoeae strain FC428 and commensal Neisseria species.

Genetic determinants of genus-level glycan diversity in a bacterial protein glycosylation system.

The human pathogens N. gonorrhoeae and N. meningitidis display robust intra- and interstrain glycan diversity associated with their O-linked protein glycosylation (pgl) systems. In an effort to better understand the evolution and function of protein glycosylation operating there, we aimed to determine if other human-restricted, Neisseria species similarly glycosylate proteins and if so, to assess the levels of glycoform diversity. Comparative genomics revealed the conservation of a subset of genes minimally required for O-linked protein glycosylation glycan and established those pgl genes as core genome constituents of the genus. In conjunction with mass spectrometric–based glycan phenotyping, we found that extant glycoform repertoires in N. gonorrhoeae, N. meningitidis and the closely related species N. polysaccharea and N. lactamica reflect the functional replacement of a progenitor glycan biosynthetic pathway. This replacement involved loss of pgl gene components of the primordial pathway coincident with the acquisition of two exogenous glycosyltransferase genes. Critical to this discovery was the identification of a ubiquitous but previously unrecognized glycosyltransferase gene (pglP) that has uniquely undergone parallel but independent pseudogenization in N. gonorrhoeae and N. meningitidis. We suggest that the pseudogenization events are driven by processes of compositional epistasis leading to gene decay. Additionally, we documented instances where inter-species recombination influences pgl gene status and creates discordant genetic interactions due ostensibly to the multi-locus nature of pgl gene networks. In summary, these findings provide a novel perspective on the evolution of protein glycosylation systems and identify phylogenetically informative, genetic differences associated with Neisseria species.

Atypical, Yet Not Infrequent, Infections with Neisseria Species.

Neisseria species are extremely well-adapted to their mammalian hosts and they display unique phenotypes that account for their ability to thrive within niche-specific conditions. The closely related species N. gonorrhoeae and N. meningitidis are the only two species of the genus recognized as strict human pathogens, causing the sexually transmitted disease gonorrhea and meningitis and sepsis, respectively. Gonococci colonize the mucosal epithelium of the male urethra and female endo/ectocervix, whereas meningococci colonize the mucosal epithelium of the human nasopharynx. The pathophysiological host responses to gonococcal and meningococcal infection are distinct. However, medical evidence dating back to the early 1900s demonstrates that these two species can cross-colonize anatomical niches, with patients often presenting with clinically-indistinguishable infections. The remaining Neisseria species are not commonly associated with disease and are considered as commensals within the normal microbiota of the human and animal nasopharynx. Nonetheless, clinical case reports suggest that they can behave as opportunistic pathogens. In this review, we describe the diversity of the genus Neisseria in the clinical context and raise the attention of microbiologists and clinicians for more cautious approaches in the diagnosis and treatment of the many pathologies these species may cause.

Children with autism spectrum disorder: Pilot studies examining the salivary microbiome and implications for gut metabolism and social behavior

IntroductionAutism Spectrum Disorder (ASD) is a collection of neurodevelopmental disorders defined by core deficits, including impaired communication, reciprocal social interaction, and stereotyped and repetitive patterns of behaviors. The salivary microbiota may serve as important indicators of oral and systemic health. In this pilot study, we identify components of the salivary microbiome in children with ASD. MethodsSaliva specimens were collected from 11 children with ASD (mean age: 10.68 years) and from 10 typically-developing individuals. Microbial DNA was extracted and utilized as templates for PCR amplification with V3-V4 16S rDNA-specific primers and high-throughput MiSeq sequencing. Taxonomic operational unit analyses and salivary microbiota profiles were conducted by LC Sciences (Houston TX); individual microbial species were further compared between children with ASD and typically-developing individuals. ResultsRothia species were found to be statistically more prevalent in children with ASD in comparison to typically-developing children (12.2-fold change; FDR p-value = 0.031). Alternately, Megasphaera, Moraxella, Neisseria, and Gemella species were all found at significantly higher levels in typically-developing children than children with ASD, displaying 39.2-, 31.9-, 18.8- and 14.0-fold differences, respectively (all with FDR p-values < 0.011). In boys with ASD, Moraxella and Neisseria species were found at significantly-higher levels compared to typically-developing counterparts, exhibiting 42.36- and 28.62-fold differences, respectively (FDR p-values of 0.011 and 0.0004). ConclusionUnderstanding the salivary microbiome in children with autism can lead to improved management of oral health and precision treatment planning. In addition, practitioners may be able to modify the oral microbiome as therapeutic regimens for ASD and other oral diseases.