A road map for in vivo evolution experiments with blood-borne parasitic microbes

Ruth Rodríguez-Pastor, Yarden Shafran, Nadav Knossow, Ricardo Gutiérrez, Shimon Harrus, Luis Zaman, Richard E. Lenski, Jeffrey E. Barrick, Hadas Hawlena*

*Corresponding author for this work

Research output: Contribution to journalReview articlepeer-review

5 Scopus citations


Laboratory experiments in which blood-borne parasitic microbes evolve in their animal hosts offer an opportunity to study parasite evolution and adaptation in real time and under natural settings. The main challenge of these experiments is to establish a protocol that is both practical over multiple passages and accurately reflects natural transmission scenarios and mechanisms. We provide a guide to the steps that should be considered when designing such a protocol, and we demonstrate its use via a case study. We highlight the importance of choosing suitable ancestral genotypes, treatments, number of replicates per treatment, types of negative controls, dependent variables, covariates, and the timing of checkpoints for the experimental design. We also recommend specific preliminary experiments to determine effective methods for parasite quantification, transmission, and preservation. Although these methodological considerations are technical, they also often have conceptual implications. To this end, we encourage other researchers to design and conduct in vivo evolution experiments with blood-borne parasitic microbes, despite the challenges that the work entails.

Original languageAmerican English
Pages (from-to)2843-2859
Number of pages17
JournalMolecular Ecology Resources
Issue number8
Early online date6 Jun 2022
StateE-pub ahead of print - 6 Jun 2022

Bibliographical note

Funding Information:
We thank B. Rosental, O. Gershoni-Yahalom, E. Bar-Zeev, and C. Brissette for their practical advice throughout the study. We also thank three reviewers for their helpful comments. This study was supported by an Israel Foundation Science grant (award 1391/15 to HH) and an Ecology of Infectious Diseases grant (award DEB-1813069 from the National Science foundation) under the auspices of U.S.-Israel Binational Science Foundation to LZ (PI), HH (PI), and JEB, REL, and SH (co-PIs). The authors declare no conflict of interest.

Publisher Copyright:
© 2022 The Authors. Molecular Ecology Resources published by John Wiley & Sons Ltd.


  • Bartonella spp.
  • experimental evolution
  • host–parasite adaptation
  • in vivo experiments
  • microbial pathogens
  • population bottlenecks
  • sequential passages
  • Bartonella spp
  • host-parasite adaptation


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