ORIGINAL RESEARCH article
Front. Microbiol.
Sec. Infectious Agents and Disease
Volume 16 - 2025 | doi: 10.3389/fmicb.2025.1644524
This article is part of the Research TopicTick-borne pathogen dynamics and disease control in AfricaView all articles
Molecular identification, genotyping and phylogenetic analysis of Ixodes and Rhipicephalus ticks and their associated spotted fever group Rickettsia species from a single location in northern Tunisia
Provisionally accepted
Myriam Kratou*
Hanène Belkahia
Rachid SelmiMeriam Ben AbdallahGhassan Tayh
Lilia Messadi
Mourad Ben Said*- National Veterinary School of Sidi Thabet, Ariana, Tunisia
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Ticks and their associated spotted fever group Rickettsia (SFGR) represent an emerging zoonotic risk in Tunisia, where data on tick species distribution and pathogen prevalence remain limited. This study specifically aimed to investigate the diversity and phylogeny of Ixodes and Rhipicephalus tick species and to identify and genetically characterize their associated SFGR species in northwestern Tunisia. Tick sampling was conducted from November 2022 to March 2023, in the Jouza district, Beja Governorate, northern Tunisia. A total of 236 ticks were collected both from vegetation using flag-dragging and manually from a red fox (Vulpes vulpes) carcass encountered opportunistically in the field. Tick species were morphologically identified and confirmed by Sanger sequencing of the mitochondrial 16S rRNA gene. Rickettsia detection was performed by nested PCR targeting the ompB gene, followed by species-level identification through sequencing of ompA and gltA partial sequences. Phylogenetic analyses were conducted to assess genetic relationships. Ticks belonged to the Ixodes ricinus complex (Ix. ricinus and Ix. inopinatus), Ix. hexagonus, and the Rhipicephalus sanguineus sensu lato complex (Rh. sanguineus sensu strict and Rh. rutilus) have been identified. Twelve and ten genotypes were recorded from 45 and 59 partial 16S rRNA mitochondrial sequences isolated from Ix. ricinus and Ix. inopinatus, respectively. Additionally, one genotype was recorded from five Rh. rutilus specimens, and four genotypes were detected among 23 Rh. sanguineus (temperate lineage) individuals. Overall, 52.1% of ticks tested positive for Rickettsia spp., with significantly higher infection rates in Ix. ricinus (75.6%, p < 0.001) and Ix. inopinatus (67.4%, p < 0.01) compared to the Rhipicephalus group (37.2%) and Ix. hexagonus, which tested negative. Three Rickettsia species were identified: R. monacensis and R. helvetica in the Ix. ricinus complex (including one co-infection), and R. massiliae exclusively in Rhipicephalus ticks. Moreover, phylogenetic analysis revealed that our tick isolates and associated Rickettsia spp. from questing ticks and the red fox clustered primarily with those from other North African and Southern European countries, suggesting trans-Mediterranean strain circulation and a potential link to wildlife reservoirs.
Keywords: Vector-borne rickettsioses, Ticks, Ixodes genus, Rhipicephalus sanguineus sensu lato, molecular identification, genetic diversity, phylogeny, Tunisia
Received: 10 Jun 2025; Accepted: 22 Jul 2025.
Copyright: © 2025 Kratou, Belkahia, Selmi, Abdallah, Tayh, Messadi and Ben Said. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Myriam Kratou, National Veterinary School of Sidi Thabet, Ariana, Tunisia
Mourad Ben Said, National Veterinary School of Sidi Thabet, Ariana, Tunisia
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