The blp Locus of Streptococcus pneumoniae Plays a Limited Role in the Selection of Strains That Can Cocolonize the Human Nasopharynx

  1. Raquel Sá-Leãoa
  1. aLaboratory of Molecular Microbiology of Human Pathogens, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal
  2. bDepartment of Pediatrics and Communicable Diseases, University of Michigan, Ann Arbor, Michigan, USA
  3. cDepartment of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan, USA
  4. dBioISI—Biosystems & Integrative Sciences Institute, Faculdade de Ciências da Universidade de Lisboa, Lisbon, Portugal
  5. eInstitute for Infection and Immunity, St. George's, University of London, London, United Kingdom
  6. fBUGS Bioscience, London Bioscience Innovation Centre, London, United Kingdom
  7. gLaboratory of Molecular Genetics, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal
  8. hLaboratory of Microbiology and Infectious Diseases, The Rockefeller University, New York, New York, USA
  1. C. A. Elkins, Editor
  1. FDA Center for Food Safety and Applied Nutrition

ABSTRACT

Nasopharyngeal colonization is important for Streptococcus pneumoniae evolution, providing the opportunity for horizontal gene transfer when multiple strains co-occur. Although colonization with more than one strain of pneumococcus is common, the factors that influence the ability of strains to coexist are not known. A highly variable blp (bacteriocin-like peptide) locus has been identified in all sequenced strains of S. pneumoniae. This locus controls the regulation and secretion of bacteriocins, small peptides that target other bacteria. In this study, we analyzed a series of cocolonizing isolates to evaluate the impact of the blp locus on human colonization to determine whether competitive phenotypes of bacteriocin secretion restrict cocolonization. We identified a collection of 135 nasopharyngeal samples cocolonized with two or more strains, totaling 285 isolates. The blp locus of all strains was characterized genetically with regard to pheromone type, bacteriocin/immunity content, and potential for locus functionality. Inhibitory phenotypes of bacteriocin secretion and locus activity were assessed through overlay assays. Isolates from single colonizations (n = 298) were characterized for comparison. Cocolonizing strains had a high diversity of blp cassettes; approximately one-third displayed an inhibitory phenotype in vitro. Despite in vitro evidence of competition, pneumococci cocolonized the subjects independently of blp pheromone type (P = 0.577), bacteriocin/immunity content, blp locus activity (P = 0.798), and inhibitory phenotype (P = 0.716). In addition, no significant differences were observed when single and cocolonizing strains were compared. Despite clear evidence of blp-mediated competition in experimental models, the results of our study suggest that the blp locus plays a limited role in restricting pneumococcal cocolonization in humans.

IMPORTANCE Nasopharyngeal colonization with Streptococcus pneumoniae (pneumococcus) is important for pneumococcal evolution, as the nasopharynx represents the major site for horizontal gene transfer when multiple strains co-occur, a phenomenon known as cocolonization. Understanding how pneumococcal strains interact within the competitive environment of the nasopharynx is of chief importance in the context of pneumococcal ecology. In this study, we used an unbiased collection of naturally co-occurring pneumococcal strains and showed that a biological process frequently used by bacteria for competition—bacteriocin production—is not decisive in the coexistence of pneumococci in the host, in contrast to what has been shown in experimental models.

FOOTNOTES

    • Received 5 April 2016.
    • Accepted 10 June 2016.
    • Accepted manuscript posted online 17 June 2016.
  • Address correspondence to Raquel Sá-Leão, rsaleao{at}itqb.unl.pt.
  • * Present address: Alexandra S. Simões, Global Health and Tropical Medicine, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisbon, Portugal; Luís A. Mendes, Department of Biology & CESAM, Universidade de Aveiro, Aveiro, Portugal.

  • Citation Valente C, Dawid S, Pinto FR, Hinds J, Simões AS, Gould KA, Mendes LA, de Lencastre H, Sá-Leão R. 2016. The blp locus of Streptococcus pneumoniae plays a limited role in the selection of strains that can cocolonize the human nasopharynx. Appl Environ Microbiol 82:5206–5215. doi:10.1128/AEM.01048-16.

  • Supplemental material for this article may be found at http://dx.doi.org/10.1128/AEM.01048-16.

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