Spiroplasma Isolated From Third-Generation Laboratory Colony Ixodes persulcatus Ticks

Area of Science:

• Microbiology
• Vector Biology
• Tick Research

Background:

• Spiroplasma are endosymbionts transmitted vertically in arthropods.
• Previous studies reported Spiroplasma in field-collected Ixodes persulcatus ticks.
• The persistence of Spiroplasma in colonized I. persulcatus remains unknown.

Purpose of the Study:

• To investigate the persistence of Spiroplasma in a laboratory colony of Ixodes persulcatus.
• To assess the prevalence of Spiroplasma in colonized I. persulcatus ticks.
• To evaluate tick cell lines for Spiroplasma isolation and cultivation.

Main Methods:

• Isolation of Spiroplasma from unfed adult I. persulcatus ticks using tick cell culture.
• Genus-specific PCR amplification of 16S rRNA and rpoB genes to detect Spiroplasma.
• Sequencing of 16S rRNA gene fragments for phylogenetic analysis.

Main Results:

• Spiroplasma was successfully isolated from ticks in the third generation of the I. persulcatus laboratory colony.
• 100% of screened I. persulcatus ticks from the colony were infected with Spiroplasma.
• The 16S rRNA sequence showed 99.8% similarity to Spiroplasma from field-collected I. persulcatus.

Conclusions:

• Spiroplasma endosymbionts maintain high prevalence in colonized I. persulcatus through at least three generations.
• Tick cell lines are valuable tools for isolating and culturing Spiroplasma.
• This study confirms the stable presence of Spiroplasma in laboratory-reared I. persulcatus.