DNA Barcoding Using 18S rRNA Gene Fragments for Identification of Tick-Borne Protists in Ticks in the Republic of Korea
Badriah Alkathiri,
Subin Lee,
KyuSung Ahn,
So Youn Youn,
Mi-Sun Yoo,
Hyang-Sim Lee,
Yun Sang Cho,
Jaeyun Jung,
Kwangwon Seo,
Soochong Kim,
Rika Umemiya-Shirafuji,
Xuenan Xuan,
Dongmi Kwak,
SungShik Shin,
Seung-Hun Lee
Affiliations
Badriah Alkathiri
College of Veterinary Medicine, Chungbuk National University, Chungbuk 28644, Republic of Korea
Subin Lee
College of Veterinary Medicine, Chungbuk National University, Chungbuk 28644, Republic of Korea
KyuSung Ahn
BIOREEDS Research Institute, College of Veterinary Medicine, Chonnam National University, Gwangju 61186, Republic of Korea
So Youn Youn
Laboratory of Parasitic and Honeybee Diseases, Bacterial Disease Division, Department of Animal & Plant Health Research, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea
Mi-Sun Yoo
Laboratory of Parasitic and Honeybee Diseases, Bacterial Disease Division, Department of Animal & Plant Health Research, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea
Hyang-Sim Lee
Laboratory of Parasitic and Honeybee Diseases, Bacterial Disease Division, Department of Animal & Plant Health Research, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea
Yun Sang Cho
Laboratory of Parasitic and Honeybee Diseases, Bacterial Disease Division, Department of Animal & Plant Health Research, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea
Jaeyun Jung
Department of Biochemistry, College of Medicine, Chungbuk National University, Chungbuk 28644, Republic of Korea
Kwangwon Seo
College of Veterinary Medicine, Chungbuk National University, Chungbuk 28644, Republic of Korea
Soochong Kim
College of Veterinary Medicine, Chungbuk National University, Chungbuk 28644, Republic of Korea
Rika Umemiya-Shirafuji
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan
Xuenan Xuan
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan
Dongmi Kwak
College of Veterinary Medicine, Kyungpook National University, Daegu 41566, Republic of Korea
SungShik Shin
BIOREEDS Research Institute, College of Veterinary Medicine, Chonnam National University, Gwangju 61186, Republic of Korea
Seung-Hun Lee
College of Veterinary Medicine, Chungbuk National University, Chungbuk 28644, Republic of Korea
The objective of this study was to evaluate the diversity and prevalence of tick-borne protists in the Republic of Korea via DNA barcoding using 18S rRNA gene fragments and PCR. Between 2021 and 2022, questing ticks were collected using the flagging method, with a total of 13,375 ticks collected and pooled into 1003 samples. Of these, 50 tick pools were selected for DNA barcoding targeting the V4 and V9 regions of 18S rRNA using the MiSeq platform. A taxonomic analysis of the amplicon sequence variants identified three genera of protozoa, namely Hepatozoon canis, Theileria luwenshuni, and Gregarine sp. However, the number and abundance of protists detected were different depending on the primer sets, and T. gondii was not identified in DNA barcoding. Furthermore, conventional PCR confirmed the presence of H. canis, Toxoplasma gondii, T. luwenshuni, and Theileria sp. in the collected ticks. This study identified H. canis and T. gondii in Ixodes nipponensis for the first time. It demonstrated that the results of DNA barcoding using 18S rRNA gene fragments can vary depending on the primer sets and further optimization is required for library construction to identify tick-borne protists in ticks. Despite these limitations, the findings highlight the potential of DNA barcoding using 18S rRNA gene fragments for screening the diversity of tick-borne protists in ticks.
