Molecular Detection of Borrelia burgdorferi s.l. (Borreliella) and Chlamydia-Like Organism DNA in Early Developmental Stages of Arthropod Vector Species

Abstract

Borrelia burgdorferi sensu lato (Bbsl) is spirochetes transmitted by ticks and known to cause Lyme disease. Chlamydia-like organisms (CLOs) comprise a large group of bacteria that can lead to serious health disorders, including miscarriage. Recently, CLOs have been found in ticks and patient skin biopsies. Due to the involvement of multiple potential vectors in the spread of these pathogens, the objective of this study was to confirm the presence of both organisms in the early developmental stages of selected vectors. Three potential vectors, Ixodes ricinus larvae, Culex pipiens larvae, and winged (unfed) adults of Lipoptena cervi, were collected in the Czech Republic in years 2019–2020. The presence of Bbsl and panchlamydial DNA was detected by PCR and positive samples were further analyzed by Sanger sequencing and phylogenetic tree construction. Bbsl DNA was proved in 1.5% (2/137) of I. ricinus larvae (identified as Borrelia afzelii and Borrelia garinii), in 1.7% (2/119) of C. pipiens larvae (both identified as B. garinii), and in 11% (3/27) of L. cervi (all identified as B. garinii). CLOs were identified in 0.7% (1/137) of I. ricinus larvae (Candidatus Protochlamydia) and in 7.4% (2/27) of L. cervi (unspecified genus), while C. pipiens larvae could not be evaluated (0%). This research represents the first investigation of the presence of CLOs in L. cervi. The detection of pathogen DNA in the early developmental stages of vectors suggests the potential for transgenerational transmission of Bbsl and CLOs in the selected vectors, although at a low rate.