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Updated: Aug 23, 2025
Tick Microbiome Characterization by Next-Generation 16S rRNA Amplicon Sequencing
Published on: August 25, 2018
Comparative analysis of microbial communities in different growth stages of Dermacentor nuttalli
Li Zhao1,2, Yi-Min Ma1, Bo Yang3
1College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, China.
Abstract:
Ticks were identified as arthropods that are pathogenic vectors. Dermacentor nuttalli is one of the dominant tick species in Inner Mongolia, and it carries and transmits a wide range of pathogenic microorganisms. However, at present, only the detection of D. nuttalli adult ticks and D. nuttalli different developmental stages carrying one specific pathogen, or the next-generation sequencing of D. nuttalli adult ticks were available. In this study, we investigated the microbial community structures of D. nuttalli in different growth stages under laboratory artificial feeding conditions. Total DNA was extracted from seven growth stages (female adult ticks, eggs, larval ticks, engorged larval ticks, nymphal ticks, engorged nymphal ticks, and second-generation adult ticks) obtained from laboratory artificial feeding of engorged D. nuttalli female ticks in Inner Mongolia. Then, the 16S rDNA V3-V4 hypervariable region was amplified to construct an Illumina PE250 library. Finally, 16S rRNA sequencing was performed on Illumina Novaseq 6000 platform. The sequencing data were analyzed using molecular biology software and platforms. The Illumina PE250 sequencing results showed that the egg stage had the highest diversity and number of species (28.74%, 98/341), while the engorged nymph stage had the lowest diversity and number of species (9.72%, 21/216). A total of 387 genera of 22 phyla were annotated in D. nuttalli, with 9 phyla and 57 genera found throughout all 7 growth stages. The dominant phylum was Proteobacteria; the dominant genera were Arsenophonus and Rickettsia; and the genera with the highest relative abundance in the 7 growth stages were Pseudomonas, Paenalcaligenes, Arsenophonus, Arsenophonus, Pseudomonas, Arsenophonus, and Rickettsia, respectively. Among the 23 exact species annotated, Brucella melitensis exhibits pathogeny that poses a serious threat to humans and animals. In this study, the microbial community composition at different growth stages of D. nuttalli was comprehensively analyzed for the first time.
Insights
This study reveals the microbial communities in all life stages of the Inner Mongolian tick, *Dermacentor nuttalli*. It identifies key bacterial genera and highlights the presence of *Brucella melitensis*, a significant pathogen.
Area of Science:
- Microbiology
- Vector-borne diseases
- Genomics
Background:
- Ticks are significant vectors of pathogenic microorganisms.
- *Dermacentor nuttalli* is a dominant tick species in Inner Mongolia, known to transmit various pathogens.
- Previous research focused on specific pathogens or adult ticks, lacking a comprehensive view of microbial communities across tick life stages.
Purpose of the Study:
- To investigate and characterize the microbial community structures in *Dermacentor nuttalli* across its seven developmental stages.
- To identify dominant microbial phyla and genera within this tick species.
- To assess the presence of potentially pathogenic microorganisms throughout the tick's life cycle.
Main Methods:
- Artificial feeding of *Dermacentor nuttalli* female ticks to obtain different life stages (eggs, larvae, nymphs, adults).
- DNA extraction from seven distinct growth stages.
- 16S ribosomal DNA (rDNA) V3-V4 hypervariable region amplification and Illumina sequencing (PE250 library, Novaseq 6000 platform).
- Bioinformatic analysis of sequencing data to determine microbial community composition.
Main Results:
- The egg stage exhibited the highest species diversity (28.74%), while the engorged nymph stage showed the lowest (9.72%).
- A total of 387 genera from 22 phyla were identified, with Proteobacteria being the dominant phylum.
- Key genera included *Arsenophonus* and *Rickettsia*. *Brucella melitensis*, a human and animal pathogen, was detected.
- Specific genera varied in abundance across life stages, with *Pseudomonas*, *Paenalcaligenes*, *Arsenophonus*, and *Rickettsia* being prominent.
Conclusions:
- This study provides the first comprehensive analysis of microbial communities across all growth stages of *Dermacentor nuttalli*.
- The findings reveal dynamic shifts in microbial composition throughout the tick's life cycle.
- The detection of *Brucella melitensis* underscores the public health importance of *Dermacentor nuttalli* as a potential vector.
