Molecular Characterization of Spiroplasma Endosymbionts in Ticks on Tortoises in Niğde Province, Türkiye

This study was conducted to detect and molecularly characterize Wolbachia and Spiroplasma endosymbionts in tortoise ticks in the Niğde province, Türkiye.

For this purpose, 184 ticks (167 males, 17 females) collected from a total of 88 tortoises (Testudo graeca) were preserved in sealed bottles containing 70% alcohol, and their species and sex were determined under a stereo microscope. Tick pools (a total of 68 pools) were designed from the 184 tick samples collected from the tortoises, with separate pools for male and female ticks from each tortoise. Following DNA extraction from the pools, PCR analyses were performed using Wolbachia and Spiroplasma-specific primers, and the genomic DNA pools were examined for amplification of the Wolbachia wsp (610 bp) and Spiroplasma 16 S rDNA (1000 bp) gene regions.

Ticks were detected on 62 of 88 tortoises (70.4%). All collected ticks were identified as adult Hyalomma aegyptium. Monthly variation in infestation rates was statistically significant (p < 0.05), whereas tick sex distribution among infested hosts was not significant (p > 0.05). PCR amplification of the wsp gene yielded no positive samples among the 68 tick pools. Only the positive control produced the expected 610 bp amplicon. Spiroplasma DNA was identified in 4/68 pools (5.8%). All positive samples originated from male ticks.

This study represents the first molecular evidence of Spiroplasma spp. (5.8%) in H. aegyptium ticks in Central Anatolia. No evidence of Wolbachia infection was detected. These findings are consistent with several recent studies reporting low or absent Wolbachia prevalence in ixodid ticks.