Comparative genomic analysis and in vivo modeling of streptococcus pneumoniae ST3081 and ST618 isolates reveal key genetic and phenotypic differences contributing to clonal replacement of serotype 1 in the Gambia

Streptococcus pneumoniae serotype 1 is one of the leading causes of invasive pneumococcal disease (IPD) in West Africa, with ST618 being the dominant cause of IPD in Te Gambia. Recently however, a rare example of clonal replacement was observed, where the ST3081 clone of serotype 1 replaced the predominant ST618 clone as the main cause of IPD. In the current study, we sought to fnd the reasons for this unusual replacement event. Using whole-genome sequence analysis and clinically relevant models of in vivo infection, we identifed distinct genetic and phenotypic characteristics of the emerging ST3081 clone. We show that ST3081 is signifcantly more virulent than ST618 in models of invasive pneumonia, and is carried at higher densities than ST618 during nasopharyngeal carriage. We also observe sequence type-specifc accessory genes and a unique sequence type-specifc fxed mutation in the pneumococcal toxin pneumolysin, which is associated with increased hemolytic activity in ST3081 and may contribute to increased virulence in this clone. Our study provides evidence that, within the same serotype 1 clonal complex, biological properties differ signifcantly from one clone to another in terms of virulence and host invasiveness, and that these differences may be the result of key genetic differences within the genome.