Skip directly to site content Skip directly to page options Skip directly to A-Z link Skip directly to A-Z link Skip directly to A-Z link
Volume 24, Number 5—May 2018
Research Letter

Borrelia miyamotoi sensu lato in Père David Deer and Haemaphysalis longicornis Ticks

Yi Yang, Zhangping Yang1Comments to Author , Patrick Kelly, Jing Li, Yijun Ren, and Chengming Wang1Comments to Author 
Author affiliations: Yangzhou University College of Veterinary Medicine, Yangzhou, China (Y. Yang, J. Li, C. Wang); Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou (Y. Yang, J. Li, C. Wang); Yangzhou University College of Animal Science and Technology, Yangzhou (Z. Yang); International Corporation Laboratory of Agriculture and Agricultural Products Safety, Yangzhou University, Yangzhou (Z. Yang); Ross University School of Veterinary Medicine, Basseterre, St. Kitts and Nevis (P. Kelly); Dafeng Elk National Natural Reserve, Yancheng, China (Y. Ren); Auburn University, Auburn, Alabama, USA (C. Wang)

Cite This Article


By sequence analysis of 16S rRNA, flaB, p66, and glpQ, we identified Borrelia miyamotoi in 1 of 4 Père David deer (n = 43) seropositive for Borrelia spp. and 1.2% (3/244) of Haemaphysalis longicornis ticks from Dafeng Elk National Natural Reserve, China. Future studies should assess Borrelia pathogenesis in deer.

Père David deer (Elaphurus davidianus) are extinct in the wild and found only in captivity, principally in China, England, and the United States. Just 5,000 animals remain, with 40% located in Dafeng Elk National Natural Reserve in China, which attracts >1 million tourists annually. Ticks are common in the Dafeng Elk National Natural Reserve (1), so we investigated the tickborne bacterial pathogens in Père David deer at this reserve.

The institutional animal care and use committee of Yangzhou University College of Veterinary Medicine (Yangzhou, China) (YZU-CVM#2015–076) approved this study. We took whole blood samples from 43 apparently healthy Père David deer (20 males, 23 females), separated out the plasma (1,800 × g for 10 min), and used the plasma to detect antibodies against bacterial pathogens with the SNAP 4Dx kit (IDEXX, Westbrook, ME, USA) (2) according to the manufacturer’s instructions. Further, ELISAs and Western blots using Borrelia miyamotoi GlpQ recombinant protein (RayBiotech, Norcross, GA, USA) and peroxidase-labeled rabbit anti-deer IgG (SeraCare, Milford, MA, USA) were performed as described previously (3) to detect GlpQ antibodies specific to B. miyamotoi.

We collected a convenience sample of Haemaphysalis longicornis ticks (n = 244) from elk in the Dafeng Elk National Natural Reserve during the summer of 2016 and stored the collection at −80°C. We used the High Pure PCR Template Preparation Kit (Roche Diagnostics GmbH, Mannheim, Germany) according to the manufacturer’s instructions to extract DNA from Père David deer whole blood samples and entire H. longicornis ticks. We used published PCR protocols targeting the 16S rRNA (4), flaB (5), and glpQ (6) genes and an in-house p66 PCR (forward primer 5′-CGATTTTTCTATATTTGGACACAT-3′, reverse primer 5′-GATATAGATTCTACAGGTATTGCATAATC-3′) to screen blood samples and ticks for B. miyamotoi. We sequenced both strands of PCR products using BGI’s (Shanghai, China) services and aligned them using ClustalW in MEGA 7 ( with the nucleotide sequences of 11 relapsing fever group borreliae and 7 Lyme disease group borreliae found in GenBank.

Four (9.3%; 1 male, 3 females) of the 43 deer were seropositive by SNAP 4Dx, demonstrating an immunodominance of antibodies against synthetic C6 peptide invariable region 6 of the pathogenic Borrelia genospecies, B. burgdorferi sensu stricto, B. garinii, and B. afzelii (7). Seropositivity was confirmed by GlpQ antibody ELISA and Western blot with GlpQ recombinant protein, indicating exposure to B. miyamotoi.


Thumbnail of Neighbor-joining phylogenetic trees constructed with 16S rRNA, flaB, p66, and glpQ gene sequences of Borrelia spp. isolates collected from Père David deer (Elaphurus davidianus) and Haemaphysalis longicornis ticks, Dafeng Elk National Natural Reserve, China, and reference isolates. The isolates identified in this study (bold; GenBank accession nos. MF521973, MF541143, MG763228, MG763229) are most similar to B. miyamotoi of the relapsing fever group. Numbers at branch nodes show boot

Figure. Neighbor-joining phylogenetic trees constructed with 16S rRNA, flaB, p66, and glpQ gene sequences of Borrelia spp. isolates collected from Père David deer (Elaphurus davidianus) and Haemaphysalis longicornis ticks, Dafeng Elk National...

One of the seropositive female deer (2.3% of overall deer population) and 3 (1.2%) of the 244 ticks were positive for the 4 Borrelia genes tested (16S rRNA, flaB, glpQ, p66) by PCR. The sequences obtained from the PCR products showed the 4 animals had identical sets of Borrelia genes. The 16S rRNA, flaB, and p66 sequences were more similar to those of the relapsing fever group borreliae (16S rRNA 97.9%–99.3%, flaB 83.7%–88.9%, p66 72.4%–83.3%) than the Lyme disease group borreliae (16S rRNA 96.6%–97.2%, flaB 79.2%–80.9%, p66 66.1%–68.1%). The B. miyamotoi glpQ gene sequence obtained from the deer and ticks also clustered with those of the relapsing fever group borreliae (81.1%–88.9%), and all analyzed gene sequences had greatest similarity with B. miyamotoi genes (16S rRNA 99.3% [576/580], flaB 88.9% [321/361], p66 83.3% [423/508], glpQ 88.9% [377/424]) (Figure).

B. miyamotoi is a member of the relapsing fever group first isolated in Japan and subsequently found in North America, Europe, and Russia (8). B. miyamotoi has not been reported in deer but can be pathogenic in humans, usually resulting in an acute febrile influenza-like illness but occasionally causing severe disease, including meningoencephalitis (9). Further studies are needed to determine the effects of B. miyamotoi infections in deer, especially because studies on Ixodes scapularis ticks in the United States have indicated that deer might be a sylvatic reservoir (10).

I. persulcatus and I. pavlovskyi ticks are known to be infected with B. miyamotoi in Asia, whereas other Ixodes spp. ticks are vectors in the United States and Europe (9). Tick control in semi–free-ranging animals is challenging; the Père David deer we studied are commonly infested with ticks. The only tick species identified on Père David deer in Dafeng Elk National Natural Reserve was H. longicornis (1), which can reach high densities in the environment (summer 89.5 ± 17.1 ticks/10 m2, winter 1.47 ± 0.35 ticks/10 m2) and cause anemia and even death in heavily infested animals. Our finding of B. miyamotoi in H. longicornis ticks adds to the list of organisms reported in this tick, primarily B. burgdorferi sensu lato and unclassified Borrelia spp.

In summary, we have shown that B. miyamotoi sensu lato occurs in Père David deer and H. longicornis ticks in Dafeng Elk National Natural Reserve. Further studies are needed on the pathogenicity of the organism in deer and the role of H. longicornis ticks in the epidemiology of infections in deer and humans.

Dr. Yang is a doctoral student at the Yangzhou University College of Veterinary Medicine in Yangzhou, China. His primary research interests are veterinary pathogens with zoonotic potential.



This project was supported by the National High Technology Research and Development Program of China (2013AA102505-5), National Natural Science Foundation of China (31472067), Scientific Innovation Research of College Graduate Program in Jiangsu Province (KYZZ16_0496), the Excellent Doctoral Dissertation Scholarship of Yangzhou University, and the Priority Academic Program Development of Jiangsu Higher Education Institutions.



  1. Shen  H, Ding  YH, Xu  AH, Ren  YJ, Hou  LB, Yuan  GX. Haemaphysalis longicornis in captive and free-ranging Père David’s deer (Elaphurus davidianus). Chin J Vet Parasitol. 2007;15:601.
  2. Chandrashekar  R, Daniluk  D, Moffitt  S, Lorentzen  L, Williams  J. Serologic diagnosis of equine borreliosis: evaluation of an in-clinic enzyme-linked immunosorbent assay (SNAP® 4Dx®). Int J Appl Res Vet Med. 2008;6:14550.
  3. Krause  PJ, Narasimhan  S, Wormser  GP, Barbour  AG, Platonov  AE, Brancato  J, et al.; Tick Borne Diseases Group. Borrelia miyamotoi sensu lato seroreactivity and seroprevalence in the northeastern United States. Emerg Infect Dis. 2014;20:118390. DOIPubMedGoogle Scholar
  4. Zhai  B, Niu  Q, Yang  J, Liu  Z, Liu  J, Yin  H, et al. Identification and molecular survey of Borrelia burgdorferi sensu lato in sika deer (Cervus nippon) from Jilin Province, north-eastern China. Acta Trop. 2017;166:547. DOIPubMedGoogle Scholar
  5. Clark  KL, Leydet  B, Hartman  S. Lyme borreliosis in human patients in Florida and Georgia, USA. Int J Med Sci. 2013;10:91531. DOIPubMedGoogle Scholar
  6. Fomenko  NV, Livanova  NN, Borgoiakov  VI, Kozlova  IV, Shulaĭkina  IV, Pukhovskaia  NM, et al. [Detection of Borrelia miyamotoi in ticks Ixodes persulcatus from Russia]. Parazitologiia. 2010;44:20111.PubMedGoogle Scholar
  7. Liang  FT, Aberer  E, Cinco  M, Gern  L, Hu  CM, Lobet  YN, et al. Antigenic conservation of an immunodominant invariable region of the VlsE lipoprotein among European pathogenic genospecies of Borrelia burgdorferi SL. J Infect Dis. 2000;182:145562. DOIPubMedGoogle Scholar
  8. Fonville  M, Friesema  IH, Hengeveld  PD, Docters van Leeuwen  A, Jahfari  S, Harms  MG, et al. Human exposure to tickborne relapsing fever spirochete Borrelia miyamotoi, the Netherlands. Emerg Infect Dis. 2014;20:12445. DOIPubMedGoogle Scholar
  9. Krause  PJ, Fish  D, Narasimhan  S, Barbour  AG. Borrelia miyamotoi infection in nature and in humans. Clin Microbiol Infect. 2015;21:6319. DOIPubMedGoogle Scholar
  10. Han  S, Hickling  GJ, Tsao  JI. High prevalence of Borrelia miyamotoi among adult blacklegged ticks from white-tailed deer. Emerg Infect Dis. 2016;22:3168. DOIPubMedGoogle Scholar




Cite This Article

DOI: 10.3201/eid2405.171355

Original Publication Date: March 26, 2018

1These senior authors contributed equally to this article.

Table of Contents – Volume 24, Number 5—May 2018

EID Search Options
presentation_01 Advanced Article Search – Search articles by author and/or keyword.
presentation_01 Articles by Country Search – Search articles by the topic country.
presentation_01 Article Type Search – Search articles by article type and issue.



Please use the form below to submit correspondence to the authors or contact them at the following address:

Zhangping Yang, Yangzhou University College of Animal Science and Technology, 12 East Wenhui Rd, Yangzhou, China, 225009; ; Chengming Wang, 1130 Wire Rd, College of Veterinary Medicine, Auburn University, Auburn, AL 36849, USA

Send To

10000 character(s) remaining.


Page created: April 17, 2018
Page updated: April 17, 2018
Page reviewed: April 17, 2018
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.