Tick-borne pathogens in ticks collected from birds in Taiwan
Kuo et al. Parasites & Vectors
Tick-borne pathogens in ticks collected from birds in Taiwan
Chi-Chien Kuo 0 1
Yi-Fu Lin 0 5
Cheng-Te Yao 4
Han-Chun Shih 3
Lo-Hsuan Chung 3
Hsien-Chun Liao 3
Yu-Cheng Hsu 2
Hsi-Chieh Wang 3
0 Equal contributors
1 Department of Life Science, National Taiwan Normal University , Taipei , Taiwan
2 Department of Natural Resources and Environmental Studies, National Dong Hwa University , Hualien , Taiwan
3 Center for Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare , Taipei , Taiwan
4 Endemic Species Research Institute, Council of Agriculture , Chi-chi, Nantou , Taiwan
5 Department of Life Science, National Chung Hsing University , Taichung , Taiwan
Background: A variety of human diseases transmitted by arthropod vectors, including ticks, are emerging around the globe. Birds are known to be hosts of ticks and can disperse exotic ticks and tick-borne pathogens. In Taiwan, previous studies have focused predominantly on mammals, leaving the role of birds in the maintenance of ticks and dissemination of tick-borne pathogens undetermined. Methods: Ticks were collected opportunistically when birds were studied from 1995 to 2013. Furthermore, to improve knowledge on the prevalence and mean load of tick infestation on birds in Taiwan, ticks were thoroughly searched for when birds were mist-netted at seven sites between September 2014 and April 2016 in eastern Taiwan. Ticks were identified based on both morphological and molecular information and were screened for potential tick-borne pathogens, including the genera Anaplasma, Babesia, Borrelia, Ehrlichia and Rickettsia. Finally, a list of hard tick species collected from birds in Taiwan was compiled based on past work and the current study. Results: Nineteen ticks (all larvae) were recovered from four of the 3096 unique mist-netted bird individuals, yielding a mean load of 0.006 ticks/individual and an overall prevalence of 0.13%. A total of 139 ticks from birds, comprising 48 larvae, 35 nymphs, 55 adults and one individual of unknown life stage, were collected from 1995 to 2016, and 11 species of four genera were identified, including three newly recorded species (Haemaphysalis wellingtoni, Ixodes columnae and Ixodes turdus). A total of eight tick-borne pathogens were detected, with five species (Borrelia turdi, Anaplasma sp. clone BJ01, Ehrlichia sp. BL157-9, Rickettsia helvetica and Rickettsia monacensis) not previously isolated in Taiwan. Overall, 16 tick species of five genera have been recorded feeding on birds, including nine species first discovered in this study. Conclusion: Our study demonstrates the paucity of information on ticks of birds and emphasizes the need for more research on ticks of birds in Taiwan and Southeast Asia. Moreover, some newly recorded ticks and tick-borne pathogens were found only on migratory birds, demonstrating the necessity of further surveillance on these highly mobile species.
Birds; Migratory birds; Ticks; Tick-borne pathogens; Taiwan
Ticks transmit the largest number of pathogens among all
arthropod disease vectors and are second only to
mosquitoes in their significance for human health. Moreover,
several tick-borne diseases are expanding rapidly, such as
anaplasmosis, babesiosis, Lyme disease, spotted fever and
tick-borne encephalitis [
Ticks typically have four life stages: egg, larva, nymph
and adult; a single blood meal from vertebrates is
necessary for the larva and nymph to molt into the next life
stage and for the adult female to lay eggs [
mammals are often one of the primary hosts for
immature ticks [
], but a growing number of studies have
revealed the significance of birds as hosts, as well as the
role migratory birds play in the long-distance
dissemination of ticks and tick-borne pathogens [
example, immature western black-legged ticks Ixodes
pacificus, the principal vector responsible for Borrelia
burgdorferi (sensu stricto) in California, can be found on
more avian than mammalian species [
]. Likewise, a
large number of bird species are hosts of Ixodes
]. Many tick species have been found on
migratory birds [
], demonstrating their ability to spread
ticks over long distances. More importantly, pathogens
transmitted by ticks might be imported through the
migration of birds. For example, the tick-borne spotted
fever group (SFG) rickettsiae have been detected in
exotic ticks recovered from migratory birds in Louisiana
 and Russia [
]. Migratory birds have also been
implicated in the spread of a variety of tick-borne diseases
by carrying pathogen-infected ticks, including Lyme
disease and Lyme borreliosis [
7, 23, 24
], babesiosis [
], anaplasmosis [
Crimean-Congo hemorrhagic fever [
Although birds play a significant role in the
subsistence of some ticks and tick-borne pathogens, their
importance varies among species. For example, birds
foraging primarily on the ground are more likely to
acquire ticks than species foraging in trees and shrubs
]. Species residing in dense oak woodland are more
frequently infested with ticks than species living in
chaparral, grass or a mixture of oak woodland and grass [
A review article found that in North America,
nonmigratory, ground-foraging birds are more likely to be
carriers of ticks, and major tick carriers are almost all
]. Similarly, reservoir competence of
Borrelia burgdorferi (s.s.), the etiological agent of Lyme
disease, also differed greatly among avian species [
Identifying these principal avian host species involved in
pathogen cycles can assist in a more effective control of
In Taiwan, tick-borne pathogens that have been isolated
from ticks include tick-borne SFG rickettsiae [
Anaplasma and Ehrlichia , Bartonella [
spp. bacteria that might cause Lyme borreliosis [
Cytauxzoon protozoans . Ticks assayed in these studies,
along with investigation or documentation of other
tickhost associations in Taiwan [
], focused predominantly
on mammals. The only study [
] that has focused on ticks
of birds covered a large geographical area (Oriental,
Palaearctic, Malagasy and Ethiopian regions) and provided
little information on tick-bird associations in Taiwan.
Knowledge of ticks on birds and the tick-borne pathogens
that these ticks can harbor remains very limited, not only in
Taiwan but also across Southeast Asia. In Malaysia, tick
infestation has been examined for seven avian species, but no
ticks have been found [
]. Similarly, 15 species of birds
have been investigated in Thailand, but only four ticks have
been collected on one avian species [
]. Information is also
needed on the role of migratory birds in the importation of
ticks and tick-borne pathogens that are likely of exotic
origin. The aim of this study is to investigate tick infestation
on birds in Taiwan, including both resident and migratory
birds, and screen for pathogens harbored by these ticks.
Ticks were collected from two sources: an opportunistic
collection of ticks from mist-netted or wounded birds and
a thorough examination of tick infestation on mist-netted
birds; the latter was implemented to reveal the prevalence
and load of tick infestation on birds. Finally, we updated
the list of hard tick species (Ixodidae) collected from birds
in Taiwan based on previous work and the current study.
Collection of ticks on birds
Ticks of birds were collected opportunistically during
two avian studies in which investigation of ectoparasites
was not the main purpose; that is, ticks were collected
when they were incidentally noticed by the researchers.
The first study took place between 1995 and 2008, with
birds being caught primarily by mist-netting around
Taiwan in preparation for voucher specimens to be
archived in the Endemic Species Research Institute of
Taiwan. The collected ticks were preserved in 75% ethanol
and were stored at room temperature. Among this
collection of ticks, 12 have been morphologically identified and
]. The other study took place during 2009–
2013, with birds being mist-netted at seven sites in the
Taroko National Park and its surrounding areas in eastern
Taiwan (Fig. 1) for a long-term avian biodiversity
monitoring project. These seven sites were (site name and
elevation in meters above sea level) as follows: Chongde
(28 m); Donghwa (41 m); Buluowan (370 m); Xibao
(980 m); Lianhua Pond (1100 m); Luoshao (1200 m); and
Hehuan Farm (2700 m) (Fig. 1). Because avian faunas vary
with elevation in Taiwan [
], these sites, with an
altitudinal difference of nearly 2700 m, covered different avian
species. These study sites also included different habitat
types, such as forests, farmlands, old fields and grassland.
At each site, 6 to 10 mist nets were erected in the
mornings and afternoons for two consecutive days, except for
Donghwa, which was surveyed for two consecutive
mornings only. Once captured, birds were banded, and a
selection of morphological characteristics was measured.
Ectoparasites, including ticks and lice, were collected
when they were incidentally noticed. Ectoparasites were
preserved in 100% ethanol and stored at -20 °C.
Because infested ticks might have been overlooked in
the above-mentioned opportunistic collections, to
improve knowledge of the prevalence (i.e. the number of
infested birds divided by the total number of the bird
individuals studied) and mean load (i.e. the total number
of ticks divided by the total number of the bird
individuals studied) of tick infestation on birds in Taiwan, from
September 2014 to April 2016, birds mist-netted at the
same seven study sites in eastern Taiwan (Fig. 1) were
thoroughly (instead of opportunistically) examined for
infestations of ticks. Ticks were also preserved in 100%
ethanol and stored at -20 °C. The number of mist nets
erected and days of survey at each study site were the
same as for the 2009–2013 period. However, the
frequency of surveys varied among the study sites due to
the difficulty of accessing some sites or less sampling
was implemented in sites with few birds trapped.
Because migratory birds stopped over in Taiwan from
September to April of the following year, the sampling
period (September 2014 to April 2016) included two
migratory seasons and a 20-month-long examination of
resident birds. Wounded birds or birds accidently
trapped by farmers during this survey period were also
carefully examined for tick infestations. In this study,
scientific names of birds and the division of migratory
and resident birds follow Clements et al. [
Identification of tick species
Ticks were examined under a dissecting microscope
(Leica MZ12) and morphologically identified to species
using published keys [
]. When unrecognized, ticks
were molecularly identified by comparing 12S rDNA and
16S rDNA sequences (primers provided in Table 1) with
known species - following Black & Piesman  and Beati
& Keirans [
]. The PCR products were purified using the
QIAquick Gel Extraction Kit (Qiagen, Valencia, USA), and
DNA sequencing was carried out using the ABI 3730XL
DNA sequencer (Applied Biosystems, Foster City, USA)
according to the manufacturer’s protocol. The PCR
products were sequenced twice in each direction and were
searched for resemblance to known tick species by using
the Basic Local Alignment Search Tool (BLAST) [
Representative sequences are submitted in the GenBank
database under accession numbers MG283136
(Haemaphysalis wellingtoni), MG283137 (Ixodes columnae), and
MG283138 (Ixodes turdus).
Pathogen identification in ticks
Because there are very few collections of ticks of birds in
Taiwan, and due to the necessity that ticks be destroyed
to be assayed for pathogen infection, only a portion of
tick samples was obtained for pathogen detection, with
another portion of tick specimens being saved as
voucher specimens. When a bird was infested with more
Real time PCR
426 or 250
rompB SFG IF
than one tick of the same species (based on
morphological identification), about half of the specimens were
selected to be assayed for pathogen infection. We also
investigated pathogens in those unrecognized ticks that
required molecular species identification.
The screened tick-borne pathogens occur in Taiwan or
in nearby countries along the bird migration routes. A
total of five groups of pathogens (and the potentially
resultant human diseases) were assayed: Anaplasma
(causative agent of anaplasmosis), Ehrlichia (causative
agent of ehrlichiosis), Rickettsia (SFG rickettsiae), Borrelia
(causative agent of Lyme disease) and Babesia (causative
agent of babesiosis). Primers and methods for amplifying
fragments of these pathogens were provided in Table 1.
The PCR protocol also followed the manufacturer’s
instructions, and positive samples were sequenced to
identify potential microbial species with a
resemblance to known species based on BLAST. Representative
sequences are submitted in the GenBank database under
accession numbers MG434346 (Borrelia turdi), MG346222
(Anaplasma sp. clone BJ01) and MG346223 (Ehrlichia sp.
Published tick species on birds in Taiwan
We compiled a list of hard ticks on birds in Taiwan based
on published papers, books, theses and the current study.
Papers were searched in PubMed (US National Library of
Medicine) and Google Scholar using the keywords “ticks”
and “Taiwan,” supplemented with a perusal of references in
these papers. We searched for potential theses with the
keyword “ticks” (both in Chinese and English) in the National
Digital Library of Theses and Dissertations in Taiwan.
Books were also searched for in Google Scholar using the
keywords “ticks” and “Taiwan” (both in Chinese and
English), and by identifying references in related papers.
When comparing frequency of tick infestation on
migratory vs resident birds, a Chi-square test was applied in
SPSS version 19.0 (Armonk, NY: IBM Corp.).
Between September 2014 and April 2016, the seven
study sites were mist-netted for a total of 56 times, with
each site surveyed 2–17 times (Chongde: 2; Donghwa:
12; Buluowan: 2; Xibao: 5; Lianhua Pond: 10; Luoshao:
17; Hehuan Farm: 8). A total of 4145 captures of 3096
individuals of 86 bird species (including 74 individuals of
wounded or accidently trapped birds) were examined for
tick infestations. Of these, 2455 individuals were
captured only once, and the remaining 641 individuals were
captured 2–8 times. These birds included 2406
individuals of 55 resident species and 690 individuals of 31
migratory species (Additional file 1: Table S1). Only larval
(but not nymphal and mature) ticks were collected,
and a total of 19 larval ticks were recovered from 4
of these 3096 bird individuals, with a mean load of
0.006 ticks/individual and a prevalence of 0.13%.
These ticks belonged to Haemaphysalis doenitzi and
Ixodes columnae, and were collected from one
resident Sinosuthora webbiana (with 14 H. doenitzi), one
resident S. webbiana (with one I. columnae), one
migratory Emberiza spodocephala (with one I. columnae)
and one migratory Turdus pallidus (with 3 I.
columnae). The mean load of ticks was the same in
resident birds (0.006 ticks/individual) as in migratory
birds (0.006 ticks/individual). Prevalence of tick
presence was more than three times higher in migratory
birds (0.29%) than in resident birds (0.08%), although
the difference was not statistically significant
(Chisquare test: χ2 = 2.0, df = 1, P = 0.16).
Overall, 139 ticks collected from birds, comprising 48
larvae, 35 nymphs, 55 adults and one individual of
unknown life stage, were examined. These included 19
larval ticks collected between September 2014 and April
2016 in eastern Taiwan, and 120 ticks opportunistically
collected from two avian studies implemented during
1995–2008 around Taiwan (83 ticks collected from 1268
bird individuals; mean load of 0.065 ticks/individual) and
2009–2013 in eastern Taiwan (37 ticks from 6343 bird
individuals; mean load of 0.006 ticks/individual). A total of
11 species of four genera (Amblyomma spp., H. doenitzi,
H. flava, H. formosensis, H. hystricis, H. ornithophila, H.
wellingtoni, I. columnae, I. granulatus, I. nipponensis, I.
turdus and Rhipicephalus haemaphysaloides) were
identified, including three newly recorded species in Taiwan (H.
wellingtoni, I. columnae and I. turdus) and six species
collected from birds in Taiwan for the first time (H. flava, H.
formosensis, H. hystricis, I. granulatus, I. nipponensis and
R. haemaphysaloides). The identity of the three newly
recorded species has been validated with 100% identity to
the nucleotide sequence deposited in GenBank (H.
wellingtoni: AB819221; I. columnae: AB819233; I. turdus:
AB819259). Ticks were collected from 19 bird species,
including seven migratory species, notably the thrush family
(Turdus chrysolaus, Turdus hortulorum, Turdus pallidus
and Zoothera dauma) (Table 2). Haemaphysalis doenitzi
was the most common species (42 ticks), comprising > 30%
of all collected ticks, followed by H. ornithophila (21 ticks),
H. wellingtoni (17 ticks) and I. columnae (17 ticks) (Table 2).
These four species accounted for nearly 70% of all
ticks. In comparison, I. columnae infested the most
diverse host species (9 species), followed by I. turdus
(4 species) and H. doenitzi (3 species) (Table 2). A few
immature ticks (16 individuals) could at best be identified to
genus (Amblyomma, Ixodes or Haemaphysalis species)
based on morphology and molecular methods (Table 2).
We found 10 papers containing information on hard
ticks of birds in Taiwan (Table 3). A total of five genera
and 16 species of hard ticks were identified based on the
current and past studies, including nine species not
previously discovered in Taiwan. The genus Haemaphysalis
(9 species) was most represented, followed by Ixodes (5
species); each of the Dermacentor and Rhipicephalus
contained one species and Amblyomma ticks could not
be identified to the species level (Table 3).
A total of 85 ticks were individually assayed for
pathogen infection, including 5 Amblyomma spp., 24 H.
doenitzi, 1 H. flava, 1 H. formosensis, 9 H. hystricis, 6 H.
ornithophila, 8 H. wellingtoni, 3 Haemaphysalis spp., 17
I. columnae, 4 I. granulatus, 1 I. nipponensis, 5 I. turdus,
and 1 R. haemaphysaloides. One Anaplasma species
(Anaplasma sp. clone BJ01), one Babesia species (Ba.
microti), two Borrelia species (Bo. valaisiana and Bo.
turdi), one Ehrlichia species (Ehrlichia sp. BL157‐9) and
three Rickettsia species (R. conorii, R. helvetica and R.
monacensis) were successfully sequenced from six tick
species (Table 4). Rickettsia helvetica, or a closely related
species, was most frequently identified (8 times, all from
the tick I. columnae), followed by Bo. valaisiana (3 times
from I. granulatus) and Bo. turdi (twice from I. turdus).
The other five pathogen species were detected only once
(Table 4). Two-thirds of the 18 detections of pathogens
were on ticks collected from migratory birds, particularly
the pale thrush (T. pallidus) (Table 4).
This is one of the few studies focusing on bird-derived
ticks and their pathogens in Southeast Asia, showing 11
tick species, of which H. wellingtoni, I. columnae and I.
turdus are new records for Taiwan, and H. flava, H.
formosensis, H. hystricis, I. granulatus, I. nipponensis and R.
haemaphysaloides have been collected from birds in
Taiwan for the first time (but had been previously
collected from mammals). In addition, eight pathogens have
been detected in these ticks, among which Bo. turdi,
Anaplasma sp. clone BJ01, Ehrlichia sp. BL157-9, R.
helvetica and R. monacensis have not previously been
identified in Taiwan. Migratory birds were found to host
infected ticks and may play a role in disseminating
pathogens. Our study demonstrates the paucity of
information on ticks of birds and emphasizes the need for more
research on ticks of birds in Taiwan.
Haemaphysalis wellingtoni, I. columnae and I. turdus
have not previously been recorded in Taiwan, and this
could be due to the limited research on ticks of birds,
rather than a rare occurrence of these tick species in
Taiwan, because birds are the primary hosts of these
three species [
] and these ticks were repeatedly
collected from birds or infested birds in large numbers
(Table 2). For example, 17 H. wellingtoni were found on
a resident, ground-foraging coucal Centropus sinensis in
a small islet (Kinmen) near mainland China. Ixodes
columnae has been found on various bird species in
Taiwan, including both resident and migratory birds.
Ixodes turdus was collected from four avian species:
three migratory birds and one resident bird. This tick
species has also been found on migratory birds in Japan
] and Korea [
], suggesting that migratory birds
can potentially disperse I. turdus across countries.
The ticks H. flava, H. formosensis, H. hystricis, I.
granulatus, I. nipponensis and R. haemaphysaloides have
previously been found feeding on mammals in Taiwan, but
not on birds. Mammals are the predominant hosts of
these six tick species, although H. flava and I. granulatus
can also be collected from birds [
]. Indeed, except for
H. hystricis and I. granulatus, the other four species were
rarely collected from birds (less than two tick
individuals) (Table 2). Less frequent occurrence on birds and
the lack of research on ticks of birds in Taiwan help
explain why these six tick species were not previously
found on birds. Haemaphysalis flava was collected from
several mammal species in Taiwan, including boars, deer
and dogs [
], and this species has been repeatedly
collected from birds in Japan, particularly the migratory
true thrushes (Turdus spp.) and buntings (Emberiza
]; likewise, we found a nymphal H. flava on a
migratory T. pallidus. Both H. formosensis and H.
hystricis have been collected from mammals in Taiwan,
particularly rodents [
]. Our study reveals that migratory
birds are also the hosts of H. formosensis; in addition, we
collected 12 immature H. hystricis from birds,
demonstrating that birds might not be occasional hosts of H.
hystricis, as previously considered [
]. Ixodes granulatus
is one of the most abundant and widespread tick species
in Taiwan and infests a diverse set of rodent species
48, 50, 65
]; in this study, five I. granulatus ticks have also
been collected from migratory birds, suggesting that birds
might also serve as major hosts of these generalist ticks. By
contrast, there are very few records of I. nipponensis in
Taiwan, with two adults recovered from cattle, and I.
nipponensis was not definitely confirmed to be native to
]. Our study verifies that I. nipponensis did
occur in Taiwan, although it was retrieved from a
migratory bird and might not have yet been established in
Taiwan. Ixodes nipponensis is distributed in temperate
broadleaf and mixed forests [
] and is a common species
in Japan [
] and Korea [
]. Taiwan’s subtropical
climate may not be ideal for the subsistence of I.
nipponensis, hence the failure to establish itself in Taiwan.
Rhipicephalus haemaphysaloides is also a common ectoparasite
on rodents of Taiwan . Similar to H. hystricis,
birds are deemed to be occasional hosts of R.
], and indeed, we found only one adult
R. haemaphysaloides on a migratory, ground-feeding
In this study, Amblyomma ticks collected from a
resident bird species could not be identified to species based
on both molecular and morphological characteristics. In
Taiwan, four Amblyomma species (A. cordiferum, A.
geoemydae, A. helvolum and A. testudinarium; [
]) have been
identified, among which, birds are known to be hosts of A.
geoemydae and A. testudinarium [
]. Whether the ticks
collected in this study belong to A. geoemydae or A.
testudinarium needs further investigation.
Abbreviations: L larva, N nymph, A adult
aThe three Ixodes granulatus were removed from three different Emberiza spodocephala
bThree of the eight Ixodes columnae were removed from the same individual Turdus pallidus
Based on published studies on ticks of birds in Taiwan,
a total of five genera and 16 species of hard ticks were
found feeding on birds according to the current and past
studies (Table 3). Among these, only seven species have
previously been documented, and nine species were first
identified in Taiwan based on this single study (Table 3).
It is thus expected that more species will be discovered
after further research on avian ectoparasites. Based on
morphological characteristics, Amblyomma spp.,
Haemaphysalis campanulata and Haemaphysalis phasiana
were reported to be collected from birds [
identified those specimens through morphology and DNA
sequences but confirmed them to be Amblyomma spp., H.
ornithophila and H. doenitzi, respectively. Haemaphysalis
campanulata and H. phasiana were thus excluded from
the final list. However, it is suspected that H. phasiana
might be synonymous to H. doenitzi (see remarks by [
Indeed, 16S rDNA sequences retrieved from the
GenBank database showed 99.5% (400/402) similarity
between H. doenitzi (GenBank: JF979402) collected in
] and H. phasiana (AB819220) collected in
]. Likewise, 16S rDNA sequences of the
eight H. doenitzi assayed in this study were 96.5–
98.5% similar to the H. phasiana sequence archived
in GenBank. Whether H. phasiana is synonymous
with H. doenitzi should be resolved when more
genetic data (e.g. 12S rDNA sequence on H. phasiana
lacking in GenBank) on both species are available.
Only 19 ticks were collected during the 2014–2016
study compared with 120 ticks collected from 1995 to
2013. A further analysis reveales that the mean load of
ticks in eastern Taiwan is the same during 2014–2016 as
for the 2009–2013 period (both mean load = 0.006 ticks/
individual), but the mean load is more than 10-fold
higher when birds were captured around Taiwan (mean
load of 0.065, during 1995–2008 period) than when
birds were captured in eastern Taiwan. The reason for
much lower mean tick loads in eastern Taiwan remains
to be investigated. In addition, only larval ticks were
found during the 2014–2016 study, whereas ticks
collected during 1995–2013 were composed primarily of
nymphs and adults (29 larvae, 35 nymphs, 55 adults).
Because nymphal and adult ticks are larger than the
larvae (which are more difficult to notice), the difference in
composition of the life stages could be due to ticks being
thoroughly searched for during 2014–2016 but only
opportunistically collected during 1995–2013. Therefore, it
should be emphasized that the 1995–2013 collection is
biased toward nymphs and adults, and occurrence of
larvae is underestimated.
Ticks are primary vectors for the five parasitic genera
]; Babesia [
]; Borrelia [
]; Rickettsia [
]) identified in ticks in this study.
Among the six tick species in which eight microbial
species have been detected (Table 4), only H. ornithophila
does not bite humans [
], so people in Taiwan are at
risk of infection from most of the identified tick-borne
pathogens. In addition, although H. ornithophila feeds
primarily on birds [
], other generalist ticks might help
bridge the pathogen transmission from birds to humans.
Among the eight pathogens detected in ticks, Ba.
microti, Bo. valaisiana and R. helvetica have been
detected in larval ticks (Table 4). Relative to R. helvetica,
which can be vertically transmitted in ticks [
transovarial transmission of Ba. microti and Bo. valaisiana has
rarely been documented in ticks [
]. This suggests
that birds might be reservoirs of Ba. microti and Bo.
valaisiana, and larval ticks can be infected when feeding
on birds. Indeed, birds were shown to be the reservoirs
of Bo. valaisiana [
]. By contrast, although Ba.
microti has been detected in ticks collected from birds
]), birds are not considered to be the reservoirs
of Ba. microti . Our study nevertheless suggests that
birds might play a role in the maintenance of Ba. microti,
although the possibility that the larval tick acquires the
protozoan via co-feeding ticks cannot be ruled out.
Five pathogens, i.e. Bo. turdi, Anaplasma sp. clone
BJ01, Ehrlichia sp. BL157-9, R. helvetica and R.
monacensis, have not previously been identified in Taiwan.
Borrelia turdi was first characterized in I. turdus on
migratory E. spodocephala in Japan [
], and later in I.
turdus and I. nipponensis on migratory birds in Korea
. This spirochete has also been detected in ticks
collected primarily from Turdus spp. birds in Europe,
including Belgium [
], Norway [
], Poland [
], Portugal [
and Spain [
]. Birds have been demonstrated to be
the reservoirs of Bo. turdi . In the current study, we
showed that Bo. turdi also occurred in Taiwan, and
similarly, the spirochete was detected in I. turdus collected
from migratory birds (T. chrysolaus and T. pallidus),
suggesting that Bo. turdi might be spread by migratory birds,
particularly the thrush. Nevertheless, although I. turdus
can infest humans [
], until now, Bo. turdi has not been
found to cause Lyme borreliosis in humans [
Anaplasma and Ehrlichia are rickettsiae belonging to
the family Anaplasmataceae and are the causative agents
of several emerging human and animal diseases [
Anaplasma sp. clone BJ01 was first isolated from
Haemaphysalis longicornis in China (GenBank: JN715833). This
bacterium is closely related to uncultured Anaplasma spp.
in Korea and USA; nevertheless, a high degree of
dissimilarity in 16S rRNA sequences with known Anaplasma
species might warrant its reclassification under a new
genus . Anaplasma phagocytophilum and a novel
Anaplasma sp. have been detected in bird tissues [
indicating that birds could potentially infect ticks with
Anaplasma. Ehrlichia sp. BL157-9 was first identified in
Hyalomma asiaticum from China [
], and was closely
related to Ehrlichia sp. ERm58 in the Ehrlichia canis group
recognized in Rhipicephalus muhsamae from Mali [
Likewise, Ehrlichia chaffeensis and an Ehrlichia species
closely related to E. canis have been detected in birds [
], implying that birds might be reservoirs of Ehrlichia.
Whether Anaplasma sp. clone BJ01 and Ehrlichia sp.
BL157-9 are pathogenic to humans remains to be
Rickettsia helvetica is a tick-borne SFG rickettsia first
characterized in Ixodes ricinus from Switzerland and
later identified in several European countries [
less reported, evidence of human or tick infection by R.
helvetica has also been found in Asia, including Japan
], Thailand [
], Laos [
] and Sakhalin Island
of Russia [
]. A strain similar to R. helvetica has also
been isolated from raccoon and sika deer in Japan [
]. In this study, a strain closely related to R. helvetica
has been repeatedly detected in I. columnae recovered
from both migratory and resident birds, demonstrating
that the potentially pathogenic R. helvetica  might
have become established in Taiwan. This should concern
physicians in Taiwan, particularly when I. columnae also
bite humans [
]. Moreover, the fact that I. columnae
has only been collected from birds in Taiwan and birds
are potential reservoirs of R. helvetica [
emphasizes the need for more research on birds, their
associated ticks and their effects on public health.
Rickettsia monacensis also belongs to SFG and was first
isolated from I. ricinus in Germany [
]. This species is
widespread in Europe and can cause disease in humans
]. Rickettsiae closely related to R. monacensis were
later identified from ticks in East and Southeast Asia,
including China [
], Korea  and Thailand [
Rickettsia monacensis has also been detected in bird tissue
]. Akin to the finding in Korea [
], this study isolated
a strain genetically close to R. monacensis from I.
nipponensis. Because I. nipponensis is distributed mainly in
temperate regions [
], and in this study, I. nipponensis was
retrieved from one migratory bird, R. monacensis was
likely imported through bird migration. Recently, severe
fever with thrombocytopenia syndrome (SFTS), an
emerging infectious disease caused by the SFTS virus
and with a high mortality rate, has been detected in
I. nipponensis in Korea [
]. The occurrence of
I. nipponensis in Taiwan, despite being rare, should
thus warrant further scrutiny.
The role of birds, particularly migratory birds, in the
spread of ticks and tick-borne pathogens has received
much more recognition in recent decades [
7, 13, 115–117
In this study, several ticks and tick-borne pathogens were
discovered for the first time in Taiwan, including some that
were found only on migratory birds (the tick I. turdus; the
pathogens Bo. turdi and Ehrlichia sp. BL157-9), although it
remains unclear if this is due to a lack of studies on
ectoparasites of birds (namely, the same ticks and
pathogens might be found in resident birds after thorough
surveillance). In addition, while some ticks and pathogens
were previously recognized in Taiwan, it is unknown
whether the same species of ticks (e.g. H. flava and H.
formosensis) and tick-borne pathogens (e.g. Ba. microti and
Bo. valaisiana) recovered from migratory birds were
acquired in Taiwan or from other countries (where ticks
acquired pathogens and then were carried by migratory birds
to Taiwan), so that a cross-country genetic mixture in ticks
and pathogens is likely to occur. Therefore, the significance
of migratory birds in the spread of exotic ticks and
tickborne pathogens in Taiwan, and whether these ticks and
pathogens can become established in Taiwan, warrants
further investigation. This is particularly true when the
majority of migrant birds found in this study forage on the
], which makes them more likely to acquire
ticks compared with birds foraging in trees or shrubs
]. Studies are also needed on the seasonality of
ticks in Taiwan to assess which life stage of which
tick species is more active during the bird migration
season (September to April of the following year) and
thus more likely to be dispersed. In eastern Taiwan, it
has been demonstrated that rodents are infested with
more R. haemaphysaloides in October and November
than in the other studied seasons, and larvae peak
from October to January , but information on the
seasonality of nearly all other tick species in Taiwan
remains very limited.
Our study demonstrates a paucity of knowledge on ticks
of birds and their associated pathogens in Taiwan and
Southeast Asia. Birds are capable of spreading ticks over
long distances; moreover, pathogens harbored by ticks
might differ when ticks were collected from birds vs
mammals, with the latter much more frequently studied
than the former in Taiwan. More research on ticks of
birds is thus warranted, which can be facilitated with the
integration of ornithologists in the studies of ticks.
Additional file 1: Table S1. Species of birds and number of captures
and unique individuals mist-netted between September 2014 and April
2016 in eastern Taiwan. (DOCX 16 kb)
We would like to thank the field workers for their help with mist-netting birds.
This study was financially supported by Taiwan Ministry of Science and
Technology (MOST 104-2621-B-003-003) and National Taiwan Normal University
(103091002) to C-CK. This article was subsidized by the National Taiwan Normal
University (NTNU), Taiwan. The bird banding program was funded by the Taroko
National Park to Y-CH.
Availability of data and materials
The data supporting the conclusions of this article are included within the
article. The tick specimens are deposited in Center for Diagnostics and
Vaccine Development, Centers for Disease Control, Taipei and Endemic
Species Research Institute, Council of Agriculture, Nantou. Representative
sequences are submitted in the GenBank database under accession numbers
MG283136 (Haemaphysalis wellingtoni), MG283137 (Ixodes columnae),
MG283138 (Ixodes turdus), MG434346 (Borrelia turdi), MG346222 (Anaplasma
sp. clone BJ01) and MG346223 (Ehrlichia sp. BL157-9).
C-CK conceived and coordinated the study. Y-CH and C-TY mist-netted birds
and collected ticks. Y-FL and H-CW identified ticks based on morphology.
H-CW, H-CS, L-HC and H-CL identified ticks based on molecular methods
and detected pathogens in ticks. C-CK wrote the manuscript. All authors read
and approved the final manuscript.
Protocols of this study were approved by Taroko National Park and Council
of Agriculture (No. TB1040000117) and all animal handling procedures met
Taiwanese legal requirements.
Consent for publication
The authors declare that they have no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations.
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