Abstract
The microbial flora associated with Hyalomma anatolicum ticks was investigated using culture-dependent (CD) and independent (next generation sequencing, NGS) methods. The bacterial profiles of different organs, development stages, sexes, and of host cattle skins were analyzed using the CD method. The egg and female gut microbiota were investigated using NGS. Fourteen distinct bacterial strains were identified using the CD method, of which Bacillus subtilis predominated in eggs, larval guts and in adult female and male guts, suggesting probable transovarial transmission. Bacillus velezensis and B. subtilis were identified in cattle skin and tick samples, suggesting that skin is the origin of tick bacteria. H.anatolicum males harbour lower bacterial diversity and composition than females. The NGS analysis revealed five different bacterial phyla across all samples, Proteobacteria contributing to >95% of the bacteria. In all, 56611sequences were generated representing 6,023 OTUs per female gut and 421 OTUs per egg. Francisellaceae family and Francisella make up the vast majority of the OTUs. Our findings are consistent with interference between Francisella and Rickettsia. The CD method identified bacteria, such B. subtilis that are candidates for vector control intervention approaches such paratransgenesis whereas NGS revealed high Francisella spp. prevalence, indicating that integrated methods are more accurate to characterize microbial community and diversity.
Highlights
Tick-borne diseases (TBDs) are imposing a growing burden for human and animal health worldwide
Bacteria were plated on brain heart infusion (BHI) agar and identified based on 16S rRNA sequencing
Two strains of P. benzoelyticum and B. licheniformis were recovered from eggs, the later one was recovered from female guts
Summary
Tick-borne diseases (TBDs) are imposing a growing burden for human and animal health worldwide. Ticks are obligate blood-feeders and can transmit to humans and animals a wide variety of pathogenic agents such as arboviruses, bacteria, and parasites. Hard ticks (Ixodidae) typically have three life stages (larva, nymph, adult) and feed on distinct host species at each developmental stage, making them important sources of zoonotic diseases [1]. Hyalomma anatolicum (Acari: Ixodidae) is the most important tick involved in transmission of the Crimean-Congo hemorrhagic fever (CCHF) virus in Iran [2,3,4]. Non-pathogenic microbes may play a role in transmission of tick-borne pathogens (TBP), with many possible consequences for both animal and human health [25]
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