Molecular Detection and Phylogeny of Tick-Borne Pathogens in Ticks Collected from Dogs in the Republic of Korea
Abstract
:1. Introduction
2. Results
2.1. Distribution of Dog Ticks in the ROK
2.2. Identification of Tick Species
2.3. Detection of Tick-Borne Pathogens in Dog Ticks
2.4. Sequencing and Phylogenetic Analysis of Tick-Borne Pathogens
3. Discussion
4. Materials and Methods
4.1. Collection of Ticks
4.2. Identification of Tick Species
4.3. Isolation of Tick Nucleic Acids
4.4. Detection of Tick-Borne Pathogens
4.5. Sequencing and Phylogenetic Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Species | Stage | Tick Pool (Number of Ticks) | Total | |||
---|---|---|---|---|---|---|
Pet Dogs | Stray Dogs | Dog Shelters 1 | Military Working Dogs | |||
Haemaphysalis longicornis | Larvae | 4 (49) | 26 (126) | 4 (166) | 0 | 34 (341) |
Nymph | 73 (606) | 51 (149) | 9 (93) | 15 (110) | 148 (958) | |
Adult | 111 (129) | 282 (295) | 29 (29) | 478 (509) | 900 (962) | |
Sub total | 188 (784) | 359 (570) | 42 (288) | 493 (619) | 1082 (2261) | |
H. flava | Larvae | 0 | 0 | 0 | 0 | 0 |
Nymph | 1 (3) | 0 | 1 (1) | 1 (3) | 3 (7) | |
Adult | 2 (2) | 0 | 0 | 1 (1) | 3 (3) | |
Sub total | 3 (5) | 0 (0) | 1 (1) | 2 (4) | 6 (10) | |
Ixodes nipponensis | Larvae | 0 | 0 | 0 | 0 | 0 |
Nymph | 0 | 0 | 0 | 0 | 0 | |
Adult | 18 (18) | 2 (2) | 1 (1) | 1 (1) | 22 (22) | |
Sub total | 18 (18) | 2 (2) | 1 (1) | 1 (1) | 22 (22) | |
Total | 209 (807) | 361 (572) | 44 (290) | 496 (624) | 1110 (2293) |
Region | Site | Species | Total (%) | ||
---|---|---|---|---|---|
Haemaphysalis longicornis | H. flava | Ixodes nipponensis | |||
Northern | Pet dogs | 552 | 4 | 12 | 568 |
Stray dogs | 189 | 0 | 1 | 190 | |
Dog shelters 1 | 100 | 0 | 0 | 100 | |
Military working dogs | 599 | 3 | 1 | 603 | |
Subtotal | 1440 (62.80) | 7 (0.31) | 14 (0.61) | 1461 (63.72) | |
Central | Pet dogs | 228 | 1 | 6 | 235 |
Stray dogs | 321 | 0 | 1 | 322 | |
Dog shelters | 188 | 1 | 1 | 190 | |
Military working dogs | 20 | 0 | 0 | 20 | |
Subtotal | 757 (33.01) | 2 (0.09) | 8 (0.35) | 767 (33.45) | |
Southern | Pet dogs | 4 | 0 | 0 | 4 |
Stray dogs | 60 | 0 | 0 | 60 | |
Dog shelters | 0 | 0 | 0 | 0 | |
Military working dogs | 0 | 1 | 0 | 1 | |
Subtotal | 64 (2.79) | 1 (0.04) | 0 | 65 (2.83) | |
Total (%) | 2261 (98.60) | 10 (0.44) | 22 (0.96) | 2293 (100) |
Scheme | Stage | Number of Tick Pools | Species | |||||
---|---|---|---|---|---|---|---|---|
Anaplasma spp. | A. platys | E. canis | E. chaffeensis | Borrelia spp. | B. gibsoni | |||
Haemaphysalis longicornis | Larvae | 34 | 3 | - | - | - | - | - |
Nymph | 148 | 5 | - | - | - | - | - | |
Adult | 900 | 14 | - | - | - | 1 | - | |
Subtotal | 1082 | 22 (2.03) | - | - | - | 1 (0.09) | - | |
H. flava | Larvae | 0 | - | - | - | - | - | - |
Nymph | 3 | - | - | - | - | - | - | |
Adult | 3 | - | - | - | - | - | - | |
Subtotal | 6 | - | - | - | - | - | - | |
Ixodes nipponensis | Larvae | 0 | - | - | - | - | - | - |
Nymph | 0 | - | - | - | - | - | - | |
Adult | 22 | 2 | - | - | - | 3 | - | |
Subtotal | 22 | 2 (9.09) | - | - | - | 3 (13.64) | - | |
Total | 1110 | 24 (2.16) | 0 | 0 | 0 | 4 (0.36) | 0 |
Region | Site | Number of Tick Pools | Species | |||||
---|---|---|---|---|---|---|---|---|
Anaplasma spp. | A. platys | E. canis | E. chaffeensis | Borrelia spp. | B. gibsoni | |||
Northern | Pet dogs | 84 | 1 (1.19) | - | - | - | 1 (1.19) | - |
Stray dogs | 169 | - | - | - | - | 1 (0.59) | - | |
Dog shelters 1 | 2 | - | - | - | - | - | - | |
Military working dogs | 482 | 11 (2.28) | - | - | - | - | - | |
Subtotal | 737 | 12 (1.63) | - | - | - | 2 (0.27) | - | |
Central | Pet dogs | 121 | 3 (2.48) | - | - | - | 2 (1.65) | - |
Stray dogs | 169 | 5 (2.96) | - | - | - | - | - | |
Dog shelters | 42 | - | - | - | - | - | - | |
Military working dogs | 13 | - | - | - | - | - | - | |
Subtotal | 345 | 8 (2.32) | - | - | - | 2 (0.58) | - | |
Southern | Pet dogs | 4 | - | - | - | - | - | - |
Stray dogs | 23 | 4 (17.39) | - | - | - | - | - | |
Dog shelters | 0 | - | - | - | - | - | - | |
Military working dogs | 1 | - | - | - | - | - | - | |
Subtotal | 28 | 4 (14.29) | - | - | - | 0 | - | |
Total (%) | 1110 | 24 (2.16) | 0 | 0 | 0 | 4 (0.36) | 0 |
Pathogens | Primers | Sequences (5′-3′) | Target Gene (bp) | PCR Conditions | References |
---|---|---|---|---|---|
Anaplasma spp. | PITA-F | GTCGAACGGATTATTCTTTA | 16S rRNA (511) | 95 °C (5 min); 37 cycles of 95 °C (30 s), 50 °C (30 s), and 72 °C (40 s); 72 °C (7 min) | [39] |
PITA-R | TTCACCTTTAACTTACCGAA | ||||
A. platys | EPLAT5 | TTTGTCGTAGCTTGCTATGAT | 16S rRNA (359) | 95 °C (5 min); 37 cycles of 95 °C (30 s), 53 °C (30 s), and 72 °C (30 s); 72 °C (7 min) | [40] |
EPLAT3 | CTTCTGTGGGTACCGTC | ||||
Ehrlichia canis | ECAN5 | GCAAATTATTTATAGCCTCTGGCTATAG | 16S rRNA (365) | 95 °C (5 min); 37 cycles of 95 °C (30 s), 56 °C (30 s), and 72 °C (30 s); 72 °C (7 min) | [41,42] |
HE3 | TTATAGGTACCGTCATTATCTTCCCTA | ||||
E. chaffeensis | HE1 | ACAATATTGCTTATAACCTTTTGGTTATA | 16S rRNA (390) | 95 °C (5 min); 37 cycles of 95 °C (30 s), 56 °C (30 s), and 72 °C (30 s); 72 °C (7 min) | |
HE3 | TTATAGGTACCGTCATTATCTTCCCTA | ||||
Borrelia spp. | B3 | GCAGCTAAGAATCTTCCGCA | 16S rRNA (714) | 95 °C (5 min); 37 cycles of 95 °C (30 s), 58 °C (30 s), and 72 °C (1 min); 72 °C (7 min) | [43] |
B6 | CAACCATGCAGCACCTGTATAT | ||||
Babesia gibsoni | PIRO-F | AGTCATATGCTTGTCTTA | 18S rRNA (500) | 95 °C (5 min); 37 cycles of 95 °C (30 s), 47 °C (30 s), and 72 °C (40 s); 72 °C (7 min) | [44] |
PIRO-R | CCATCATTCCAATTACAA |
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Truong, A.-T.; Noh, J.; Park, Y.; Seo, H.-J.; Kim, K.-H.; Min, S.; Lim, J.; Yoo, M.-S.; Kim, H.-C.; Klein, T.A.; et al. Molecular Detection and Phylogeny of Tick-Borne Pathogens in Ticks Collected from Dogs in the Republic of Korea. Pathogens 2021, 10, 613. https://doi.org/10.3390/pathogens10050613
Truong A-T, Noh J, Park Y, Seo H-J, Kim K-H, Min S, Lim J, Yoo M-S, Kim H-C, Klein TA, et al. Molecular Detection and Phylogeny of Tick-Borne Pathogens in Ticks Collected from Dogs in the Republic of Korea. Pathogens. 2021; 10(5):613. https://doi.org/10.3390/pathogens10050613
Chicago/Turabian StyleTruong, A-Tai, Jinhyeong Noh, Yeojin Park, Hyun-Ji Seo, Keun-Ho Kim, Subin Min, Jiyeon Lim, Mi-Sun Yoo, Heung-Chul Kim, Terry A. Klein, and et al. 2021. "Molecular Detection and Phylogeny of Tick-Borne Pathogens in Ticks Collected from Dogs in the Republic of Korea" Pathogens 10, no. 5: 613. https://doi.org/10.3390/pathogens10050613