Identification of integrative and conjugative elements in pathogenic and commensal Neisseriaceae species via genomic distributions of DNA uptake sequence dialects.

Mobile genetic elements (MGEs) are key factors responsible for dissemination of virulence determinants and antimicrobial-resistance genes amongst pathogenic bacteria. Conjugative MGEs are notable for their high gene loads donated per transfer event, broad host ranges and phylogenetic ubiquity amongst prokaryotes, with the subclass of chromosomally inserted integrative and conjugative elements (ICEs) being particularly abundant. The focus on a small number of model systems has biased the study of ICEs towards those conferring readily selectable phenotypes to host cells, whereas the identification and characterization of integrated cryptic elements remains challenging. Even though antimicrobial resistance and horizontally acquired virulence genes are major factors aggravating neisserial infection, conjugative MGEs of Neisseria gonorrhoeae and Neisseria meningitidis remain poorly characterized. Using a phenotype-independent approach based on atypical distributions of DNA uptake sequences (DUSs) in MGEs relative to the chromosomal background, we have identified two groups of chromosomally integrated conjugative elements in Neisseria: one found almost exclusively in pathogenic species possibly deriving from the genus Kingella, the other belonging to a group of Neisseria mucosa-like commensals. The former element appears to enable transfer of traditionally gonococcal-specific loci such as the virulence-associated toxin-antitoxin system fitAB to N. meningitidis chromosomes, whilst the circular form of the latter possesses a unique attachment site (attP) sequence seemingly adapted to exploit DUS motifs as chromosomal integration sites. In addition to validating the use of DUS distributions in Neisseriaceae MGE identification, the >170 identified ICE sequences provide a valuable resource for future studies of ICE evolution and host adaptation.

An interaction between host and microbe genotypes determines colonization success of a key bumble bee gut microbiota member.

There has been a proliferation of studies demonstrating an organism's health is influenced by its microbiota. However, factors influencing beneficial microbe colonization and the evolution of these relationships remain understudied relative to host-pathogen interactions. Vertically transmitted beneficial microbes are predicted to show high levels of specificity in colonization, including genotype matching, which may transpire through coevolution. We investigate how host and bacterial genotypes influence colonization of a core coevolved microbiota member in bumble bees. The hindgut colonizing Snodgrassella alvi confers direct benefits, but, as an early colonizer, also facilitates the further development of a healthy microbiota. Due to predominantly vertical transmission promoting tight evolution between colonization factors of bacteria and host lineages, we predict that genotype-by-genotype interactions will determine successful colonization. Germ-free adult bees from seven bumble bee colonies (host genotypic units) were inoculated with one of six genetically distinct strains of S. alvi. Subsequent colonization within host and microbe genotypes combinations ranged from 0 to 100%, and an interaction between host and microbe genotypes determined colonization success. This novel finding of a genotype-by-genotype interaction determining colonization in an animal host-beneficial microbe system has implications for the ecological and evolutionary dynamics of host and microbe, including associated host-fitness benefits.

Laribacter hongkongensis: an emerging pathogen of infectious diarrhea.

Laribacter hongkongensis is relatively a new name in the list of bacterial pathogens for gastroenteritis and travelers' diarrhea. Addition of another name increases burden on the enteric infections as a whole. L. hongkongensis belongs to Neisseriaceae family of ß subclass Proteobacteria. L. hongkongensis was initially isolated in Hong Kong from blood and empyema of an alcoholic cirrhotic patient in 2001, followed by reports from Korea and China, representing a total of 38 articles in PubMed until April 2013. As of now, there is no report from Indian subcontinent where infectious diarrhea is very much prevalent and a major burden. This review provides information about the microbiological characteristics, consideration of an emerging pathogen, relative pathogenicity, genome and proteome content, resistance toward multiple antibiotics, adaptability to different stress, and other features since its time of discovery. Investigation for this bacterium may avoid misidentification as other microbial flora. Further studies like the geographical distribution, type of infection, disease burden, pathogenicity, or genomic exploration of this bacterium will be useful in characterizing them properly. This bacterium may possibly be the emerging threat to public health.

Sequence-based predictions of lipooligosaccharide diversity in the Neisseriaceae and their implication in pathogenicity.

Endotoxin [Lipopolysaccharide (LPS)/Lipooligosaccharide (LOS)] is an important virulence determinant in gram negative bacteria. While the genetic basis of endotoxin production and its role in disease in the pathogenic Neisseria has been extensively studied, little research has focused on the genetic basis of LOS biosynthesis in commensal Neisseria. We determined the genomic sequences of a variety of commensal Neisseria strains, and compared these sequences, along with other genomic sequences available from various sequencing centers from commensal and pathogenic strains, to identify genes involved in LOS biosynthesis. This allowed us to make structural predictions as to differences in LOS seen between commensal and pathogenic strains. We determined that all neisserial strains possess a conserved set of genes needed to make a common 3-Deoxy-D-manno-octulosonic acid -heptose core structure. However, significant genomic differences in glycosyl transferase genes support the published literature indicating compositional differences in the terminal oligosaccharides. This was most pronounced in commensal strains that were distally related to the gonococcus and meningococcus. These strains possessed a homolog of heptosyltransferase III, suggesting that they differ from the pathogenic strains by the presence a third heptose. Furthermore, most commensal strains possess homologs of genes needed to synthesize lipopolysaccharide (LPS). N. cinerea, a commensal species that is highly related to the gonococcus has lost the ability to make sialyltransferase. Overall genomic comparisons of various neisserial strains indicate that significant recombination/genetic acquisition/loss has occurred within the genus, and this muddles proper speciation.

Protein secretion and secreted proteins in pathogenic Neisseriaceae.

Secreted proteins of pathogenic bacteria are often essential virulence factors. They are involved, for example, in the adherence of the bacteria to host cells or required to suppress the host's defence mechanisms. Until recently, only IgA1 protease had been studied in detail in the NeisseriaceaeNeisseria meningitidis and Neisseria gonorrhoeae. The availability of their genome sequences, however, has boosted research in this area. Here, we present a survey of the secretome of the pathogenic Neisseriaceae, based on the available genome sequences, and the current knowledge of the functions and structures of the secreted proteins. Of the six protein-secretion pathways that are widely disseminated among Gram-negative bacteria, three pathways appear to be present among the Neisseriaceae, i.e. the autotransporter-, the two-partner- and the type I-secretion mechanisms. Comparison of the predicted secretomes reveals a considerable flexibility. As compared with N. meningitidis and the nonpathogen N. lactamica, N. gonorrhoeae appears to have a considerably degenerated secretome, which may reflect its altered niche occupancy. The flexibility of the secretome may be enhanced by the presence of ORFs in the genomes potentially encoding fragments of secreted proteins. We hypothesize that these ORFs may substitute for the corresponding fragments in the full-length genes through genetic recombination, thereby changing the host-cell receptor specificity of the secreted protein.

Ecoepidemiology of Laribacter hongkongensis, a novel bacterium associated with gastroenteritis.

In a territory-wide surveillance study, Laribacter hongkongensis was isolated solely from freshwater fish (60% of grass carps, 53% of bighead carps, and 25% of mud carps). Comparing the pulsed-field gel electrophoresis patterns of fish and patient isolates revealed that most patient isolates were clustered together, suggesting that some clones could be more virulent.

[Dog bite infections associated with CDC group EF-4a. Report of 2 cases]. / Infecciones por mordedura de perro asociadas a CDC grupo EF-4a. Comunicación de 2 casos.

BACKGROUND: Group EF-4 bacteria make up part of the normal flora of the oral cavity of dogs and cats. Few reports have been published on the incidence of human infections by this group of bacteria and these are associated with animal bite or scratch. Two cases of infections by CDC group EF-4 by dog bite were diagnosed in 1996 by the Bacteriology Laboratory of the authors' hospital. These cases are herein described and the biochemical analysis and profile of sensitivity of this little known group of bacteria evaluated. METHODS: Two clinical cases of infection by CDC group EF-4a by dog bite are described. Identification of the bacteria was performed by conventional biochemical tests and quantitative antibiotic sensitivity to 12 antibiotics was carried out by the seried broth macrodilution method. RESULTS: The two strains isolated corresponded to biovar "a" of group EF-4 being sensitive to: ampicillin, ceftriaxone, aminoglycosides, chloramphenicol, rifamipicin, TMS and ciprofloxacin, intermediate sensitivity to erythromycin and were resistant to cefalotine, oxacillin and vancomycin. With respect to penicillin, one of the strains was sensitive and the other presented intermediate sensitivity. Neither of the strains produced beta lactamase. CONCLUSIONS: Although Pasteurella sp. is usually considered in dog bite wounds, the possible presence of group EF-4 should be taken into account since the sensitivity of both microorganisms against penicillin and cefalotin, which are effective against Pasteurella but less active against group EF-4 bacteria differ.

Kingella kingae, a rare cause of bacterial meningitis.

A male adolescent with a history of pharyngitis developed meningitis due to Kingella kingae. This is a Gram-negative coccobacillus belonging to the family of Neisseriaceae. It is a rarely reported human pathogen, from which only 2 cases of meningitis have been described up to the present day. Our patient developed ophthalmoplegia, suggestive of basal meningitis. He was treated with penicillin G and recovered completely.

Structural heterogeneity of the lipopolysaccharides of the Neisseriaceae.

Lipopolysaccharides from 5 different genera of the Neisseriaceae were analyzed on sodium dodecylsulfate-polyacrylamide gel electrophoresis, and visualized by silver staining. Significant heterogeneity in the banding patterns was observed with some of the strains producing only low molecular mass molecules and others producing O-repeating units. All genera examined except Branhamella contained strains that were able to produce an O-repeating side chain on their lipopolysaccharides. The ability to produce the repeating subunit did not correlate with the presence of plasmids.

Empyema caused by Kingella denitrificans and Peptostreptococcus spp. in a patient with bronchogenic carcinoma.

Empyema caused by Kingella denitrificans and Peptostreptococcus spp. was diagnosed in a patient with bronchogenic carcinoma. This appears to be the third report providing evidence of a pathogenic role for K. denitrificans, and the first concerning infection in the pleural space and in a patient with underlying immunosuppressive disease. K. denitrificans should be added to the list of fastidious gram-negative bacteria associated with opportunistic infections in the compromised host.

Kingella kingae infection in healthy children.

Kingella kingae is a gram-negative occasional, but normal, inhabitant of the nasopharynx. We present two new cases of this infection that occurred in previously healthy children, and compare and contrast them to other cases reported in the literature. K. kingae osteomyelitis generally has an insidious, subacute onset, whereas septic arthritis has an acute presentation. To date, all strains of K. kingae have been sensitive to penicillin, and no residual damage has been reported following osteomyelitis or septic arthritis, except that residual disk space narrowing did occur after K. kingae discitis.

Otitis media and sinusitis in children. Role of Branhamella catarrhalis.

Otitis media and sinusitis are among the most common diseases of childhood and various treatment approaches have been advocated. This review focuses on current experience in Pittsburgh, both in terms of our methods of patient management and the changing aetiology of the underlying diseases. Special reference is made to the role played by Branhamella catarrhalis and the emergence of beta-lactamase-producing strains. Results from recently completed clinical trials would suggest that antibacterial therapy should be administered to children with these infections, to reduce morbidity and prevent the ever-present supportive complications associated with both diseases.

Branhamella catarrhalis as an indirect pathogen.

In 7 patients with bronchopulmonary infections, treatment with penicillin antibiotics failed to eradicate Haemophilus influenzae or Streptococcus pneumoniae which were sensitive to the antibiotic used. The sputum of these patients contained both the pathogen and beta-lactamase-producing Branhamella catarrhalis. No antibacterial activity was detectable in sputum samples obtained from patients receiving a penicillin antibiotic. This suggests that B. catarrhalis can act as an indirect pathogen, protecting accompanying pathogens from antibiotics by producing an antibiotic-destroying enzyme.