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Cultivation Methods of Spirochetes from Borrelia burgdorferi Sensu Lato Complex and Relapsing Fever Borrelia
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Cultivation Methods of Spirochetes from Borrelia burgdorferi Sensu Lato Complex and Relapsing Fever Borrelia

Published on: November 25, 2022

Relapsing fever spirochetes retain infectivity after prolonged in vitro cultivation

Job E Lopez1, Merry E Schrumpf, Sandra J Raffel

  • 1Laboratory of Zoonotic Pathogens, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana 59840, USA. lopezjob@niaid.nih.gov

Vector Borne and Zoonotic Diseases (Larchmont, N.Y.)
|July 22, 2008
PubMed

Insights

Relapsing fever spirochetes, Borrelia hermsii and Borrelia turicatae, maintained infectivity and genomic stability after one year of in vitro cultivation. Loss of a specific plasmid segment did not affect infectivity in mice.

Area of Science:

  • Microbiology
  • Genetics
  • Infectious Diseases

Background:

  • Borrelia hermsii and Borrelia burgdorferi are related spirochetes causing tick-borne relapsing fever and Lyme disease.
  • In vitro cultivation of B. burgdorferi leads to loss of infectivity and plasmids.
  • Effects of long-term in vitro cultivation on B. hermsii genomic stability and infectivity were previously unknown.

Purpose of the Study:

  • To analyze the impact of continuous in vitro cultivation on the genomic composition and infectivity of Borrelia hermsii and Borrelia turicatae.
  • To investigate the genetic stability of relapsing fever spirochetes during prolonged in vitro culture.

Main Methods:

  • Cultivation of seven Borrelia hermsii and one Borrelia turicatae isolates in vitro for one year.
  • Assessment of infectivity in mice after long-term cultivation.
  • Analysis of plasmid profiles and genomic sequencing, focusing on the fhbA gene.

Main Results:

  • All examined Borrelia hermsii and Borrelia turicatae isolates retained infectivity in mice after one year of in vitro cultivation.
  • Minimal changes were observed in plasmid profiles following extended cultivation.
  • Two B. hermsii isolates lost a portion of the 200-kb linear plasmid containing the fhbA gene but remained infective.

Conclusions:

  • Relapsing fever spirochetes demonstrate significant genetic stability during prolonged in vitro cultivation.
  • The fhbA-containing DNA segment lost during cultivation is not essential for Borrelia hermsii infectivity in mice.
  • These findings are crucial for understanding spirochete pathogenic mechanisms and in vitro culture methodologies.

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