Evolution of Burkholderia pseudomallei in Recurrent Melioidosis

Burkholderia pseudomallei, the etiologic agent of human melioidosis, is capable of causing severe acute infection with overwhelming septicemia leading to death. A high rate of recurrent disease occurs in adult patients, most often due to recrudescence of the initial infecting strain. Pathogen persistence and evolution during such relapsing infections are not well understood. Bacterial cells present in the primary inoculum and in late infections may differ greatly, as has been observed in chronic disease, or they may be genetically similar. To test these alternative models, we conducted whole-genome comparisons of clonal primary and relapse B. pseudomallei isolates recovered six months to six years apart from four adult Thai patients. We found differences within each of the four pairs, and some, including a 330 Kb deletion, affected substantial portions of the genome. Many of the changes were associated with increased antibiotic resistance. We also found evidence of positive selection for deleterious mutations in a TetR family transcriptional regulator from a set of 107 additional B. pseudomallei strains. As part of the study, we sequenced to base-pair accuracy the genome of B. pseudomallei strain 1026b, the model used for genetic studies of B. pseudomallei pathogenesis and antibiotic resistance. Our findings provide new insights into pathogen evolution during long-term infections and have important implications for the development of intervention strategies to combat recurrent melioidosis

Microbial Community-Driven Etiopathogenesis of Peri-Implantitis

Osseointegrated dental implants are a revolutionary tool in the armament of reconstructive dentistry, employed to replace missing teeth and restore masticatory, occlusal, and esthetic functions. Like natural teeth, the orally exposed part of dental implants offers a pristine nonshedding surface for salivary pellicle-mediated microbial adhesion and biofilm formation. In early colonization stages, these bacterial communities closely resemble those of healthy periodontal sites, with lower diversity. Because the peri-implant tissues are more susceptible to endogenous oral infections, understanding of the ecological triggers that underpin the microbial pathogenesis of peri-implantitis is central to developing improved prevention, diagnosis, and therapeutic strategies. The advent of next-generation sequencing (NGS) technologies, notably applied to 16S ribosomal RNA gene amplicons, has enabled the comprehensive taxonomic characterization of peri-implant bacterial communities in health and disease, revealing a differentially abundant microbiota between these 2 states, or with periodontitis. With that, the peri-implant niche is highlighted as a distinct ecosystem that shapes its individual resident microbial community. Shifts from health to disease include an increase in diversity and a gradual depletion of commensals, along with an enrichment of classical and emerging periodontal pathogens. Metatranscriptomic profiling revealed similarities in the virulence characteristics of microbial communities from peri-implantitis and periodontitis, nonetheless with some distinctive pathways and interbacterial networks. Deeper functional assessment of the physiology and virulence of the well-characterized microbial communities of the peri-implant niche will elucidate further the etiopathogenic mechanisms and drivers of the disease. </jats:p

Purine-containing compounds, including cyclic adenosine 3',5'-monophosphate, induce fruiting of Myxococcus xanthus by nutritional imbalance

Induction of Myxococcus xanthus fruiting by a number of different purine-containing compounds, including cyclic adenosine 3',5'-monophosphate, is defective in a mutant resistant to 2,6-diaminopurine. Furthermore, the purine-induced fruiting of wild-type cultures is uniquely blocked by a low concentration of added glycine. These results imply that different purine-containing compounds induce fruiting through a single mechanism involving nutritional imbalance.</jats:p

Guanosine pentaphosphate and guanosine tetraphosphate accumulation and induction of Myxococcus xanthus fruiting body development

Development of multicellular fruiting bodies of Myxococcus xanthus can be induced by limitation of any of a number of different classes of amino acids. Investigated were amino acids that wild-type strains of M. xanthus are unable to synthesize (isoleucine, leucine, and valine), can synthesize at a low rate (phenylalanine), or can normally synthesize at an adequate rate (tryptophan and serine). In general, gradual rather than abrupt starvation for an essential amino acid was required for the induction of fruiting. Perhaps gradual starvation in general minimizes antagonism between amino acids present in the medium, as was documented for valine starvation. The previously reported induction of fruiting by a high concentration of threonine was shown to be specifically reversed by lysine. Threonine addition may starve cells for lysine by feedback inhibition of aspartokinase activity. Starvation for carbon-energy sources or inorganic phosphate also induced fruiting. As in other bacteria, amino acid starvation of M. xanthus leads to increases in cellular guanosine polyphosphate, usually consisting of large increases in the amount of guanosine pentaphosphate with smaller increases in the level of guanosine tetraphosphate. Guanosine polyphosphate accumulation is thus shown to be correlated with nutritional conditions that induce fruiting, and therefore may serve as an intracellular signal to trigger cells to end vegetative growth and initiate fruiting body development.</jats:p

Accumulation of guanosine tetraphosphate and guanosine pentaphosphate in Myxococcus xanthus during starvation and myxospore formation

Cultures of Myxococcus xanthus develop multicellular fruiting bodies when starved for carbon and nitrogen sources on an agar surface. Under these conditions of severe starvation, cultures rapidly accumulated a compound identified as guanosine tetraphosphate by chromatographic migration of the compound and of its major acid and alkali breakdown products. The accumulation of guanosine tetraphosphate was reduced in the presence of tetracycline, indicating that it may be synthesized by mechanisms similar to those of Escherichia coli. The guanosine tetraphosphate level was also reduced in starved cultures of a mutant unable to fruit normally, although it has been determined whether the defect in guanosine tetraphosphate accumulation is responsible for the inability to fruit. Induction of spores by glycerol addition led to transient increases in both guanosine tetraphosphate and guanosine pentaphosphate at a stage following most cell shortening, but before spores had acquired full refractility.</jats:p