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Symbioses between animals and bacteria are ubiquitous. To better understand these relationships, it is essential to unravel how bacteria evolve to colonize hosts. Previously, we serially passaged the free-living bacterium,  Shewanella oneidensis , through the digestive tracts of germ-free larval zebrafish ( Danio rerio ) to uncover the evolutionary changes involved in the initiation of a novel symbiosis with a vertebrate host. After 20 passages, we discovered an adaptive missense mutation in the  mshL  gene of the  msh  pilus operon, which improved host colonization, increased swimming motility, and reduced surface adhesion. In the present study, we determined that this mutation was a loss-of-function mutation and found that it improved zebrafish colonization by augmenting  S. oneidensis  representation in the water column outside larvae through a reduced association with environmental surfaces. Additionally, we found that strains containing the  mshL  mutation were able to immigrate into host digestive tracts at higher rates per capita. However, mutant and evolved strains exhibited no evidence of a competitive advantage after colonizing hosts. Our results demonstrate that bacterial behaviors outside the host can play a dominant role in facilitating the onset of novel host associations.

Msh Pilus Mutations Increase the Ability of a Free-Living Bacterium to Colonize a Piscine Host