Abstract:
Ticks, well-known as hematophagous vectors, play a crucial role in transmitting numerous
zoonotic pathogens ranging from viruses and bacteria to protozoa. To prevent and understand
the transition from tick-borne disease (TBD) endemicity to epidemicity, it is imperative to
focus on epidemiological aspects to develop robust surveillance methods and strategies. In this
study, we developed a Nanopore-based metabarcoding approach for bacteria detection in ticks
collected from Catalunya and Valencia. A total of 186 adult ticks belonging to Rhipicephalus
bursa, Dermacentor marginatus, Hyalomma lusitanicum, Hyalomma marginatum, and Ixoded
ricinus species were studied. In this study, we used PCR amplification combined with
Nanopore sequencing techniques and bioinformatics pipelines to characterize the tick-
associated microbiota studies. Full-length 16S rRNA gene metabarcoding sequencing
demonstrated 59 species and 336 genera generated belonging to 22 phyla, 33 classes, 111
orders, and 178 families. Within this examination, we identified some novel endosymbionts
bacterial species that according to the best of our knowledge were not documented previously
in Spain including Candidatus Coxiella mudrowiae, Francisella persica, Rickettsia africae,
Rickettsia japonica, Rickettsia rhipicephali, and Unclassified Aeromonas. Candidatus Coxiella
mudrowiae and Rickettsia africae species predominated in the Rhipicephalus bursa tick. The
Francisella genus and Rickettsia rhipicephali species dominated in Hyalomma marginatum.
While Rickettsia japonica is associated with all the examined tick samples except
Rhipicephalus bursa. The alpha diversity results, as assessed by the Shannon index (p-value >
0.05), indicate that there are no statistically significant variations in the bacterial communities
within ticks, whether we consider different geographic locations or the feeding status of the
ticks.
