Tick-borne pathogens and the vector potential of ticks in China.

Ticks, as obligate blood-sucking ectoparasites, attack a broad range of vertebrates and transmit a great diversity of pathogenic microorganisms. They are considered second only to mosquitoes as vectors of human disease, and the most important vector of pathogens of domestic and wild animals. Of the 117 described species in the Chinese tick fauna, 60 are known to transmit one or more diseases: 36 species isolated within China and 24 species isolated outside China. Moreover, 38 of these species carry multiple pathogens, indicating the potentially vast role of these vectors in transmitting pathogens. Spotted fever is the most common tick-borne disease, and is carried by at least 27 tick species, with Lyme disease and human granulocytic anaplasmosis ranked as the second and third most widespread tick-borne diseases, carried by 13 and 10 species, respectively. Such knowledge provides us with clues for the identification of tick-associated pathogens and suggests ideas for the control of tick-borne diseases in China. However, the numbers of tick-associated pathogens and tick-borne diseases in China are probably underestimated because of the complex distribution and great diversity of tick species in this country.

Review

Ticks, as obligate blood-sucking ectoparasites, attack a broad range of vertebrates, including humans, and they are considered second only to mosquitoes as vectors of human disease, and the most important vector of pathogens of domestic and wild animals [1]. They transmit a variety of pathogens of medical and veterinary interest, including viruses, bacteria, rickettsiae, helminthes, and protozoans, all of which are able to cause damage to livestock production and human health. The global threat of tick-borne diseases is increasing, with new pathogens identified continuously [2]. There are an estimated 899 species of ticks belonging to three families: Argasidae, Ixodidae, and Nuttalliellidae (represented by a monotypic species restricted to South Africa) [3].

In China, 117 species of the following genera have been identified: Argas (seven species), Carios (four species), and Ornithodoros (two species) in the family Argasidae; and Amblyomma (eight species), Anomalohimalaya (two species), Dermacentor (twelve species), Haemaphysalis (forty four species), Hyalomma (six species), Ixodes (twenty four species), and Rhipicephalus (eight species) in the family Ixodidae [4]. Some of these species carry or transmit one or more infectious pathogens, resulting in severe zoonotic diseases. The most commonly observed human tick-borne diseases in China are reportedly Lyme disease, tick-borne encephalitis, Crimean-Congo hemorrhagic fever, Q fever, tularemia, and North-Asia tick-borne spotted fever [5]. Epidemiologically important tick-borne diseases, such as Human Granulocytic Anaplasmosis (HGA) and severe Fever with Thrombocytopenia Syndrome (FLTS), have also emerged in recent years. The characterization of a new bunyavirus (associated with fever, thrombocytopenia, and leukopenia syndrome) in 2010 has prompted greater attention to ticks and tick-borne diseases throughout China. However, tick-associated pathogens and diseases are still underestimated because of the complex distribution and the large diversity of tick species in China.

Although the rapid development of molecular techniques has greatly advanced the identification of emerging tick pathogens, continuous research is required to fully comprehend the diversity of tick-borne pathogens and to completely identify the vector roles of ticks in China. In this study, with regard to the Chinese tick fauna, we reviewed the tick-associated pathogenic microorganisms that have been identified world-wide, and evaluated the potential roles of the ticks as vectors throughout China. This will extend the identification of tick-associated pathogens and suggest better strategies for the control of tick-borne diseases in China.

Role of argasid ticks as vectors in China and their associated tick-borne pathogens

In China, there are 13 species of argasid ticks, belonging to three genera: Argas (seven species), Carios (four species), and Ornithodoros (two species) [4]. The majority of these are nidicolous, usually residing in the burrows, caves, or nests of their hosts. Among all the argasids found in China, four Argas species, two Carios species, and two Ornithodoros species are competent to transmit or cause human disease (Table 1) [6-17]. Among these eight tick species, four (A. japonicas, A. persicus, O. tartakovskyi, and O. tholozani) have been confirmed as causing host illnesses in China. A case of human dermatitis was recorded in 1986 after a bite by A. japonicas, but no pathogen has been identified from this tick species in China [6]. The tick A. persicus mainly infests poultry and carries the most diverse array of pathogens in the family Agarsidae, including Borrelia anserine, Kyasanur Forest disease virus, and Wolbachia persica n. sp. However, only B. anserine, known to cause avian spirochetosis, has been confirmed in China [7]. Lake Clarendon virus was isolated from A. robertsi; Quaranfil virus and Gissar virus were identified in A. vulgaris; and “Issyk-Kul” virus has been identified in C. vespertilionis. No virus has been detected in ticks collected in China. The symptoms or diseases caused by these viruses are still unclear [11-13,16], and the vector roles of these ticks in China remain unknown. Carios capensis can be coinfected by pathogen DNA from Borrelia, Coxiella, and Rickettsia, as well as West Nile virus [14,15], although no pathogens have been reported in this tick species collected in China. Ornithodoros tartakovskyi and O. tholozani both cause tick-borne relapsing fever in China, but carry different pathogens, B. latyshevyi and B. persica, respectively [17].

Table 1

Tick-borne pathogens and the vector role of argasid ticks distributed in China

Tick species	Pathogens	Diseases	References
Argas
A. japonicas	Unidentified	Dermatitis	[6]
A. persicus	Borrelia anserine; Kyasanur Forest disease virusa; Wolbachia persica, n. sp.a	Avian spirochetosis; Kyasanur forest disease; Paralysis	[7-10]
A. robertsi	Lake Clarendon virusa	unknown	[11]
A. vulgaris	Quaranfil virusa; Gissar virusa	unknown	[12,13]
Carios
C. capensis	West Nile Virusa,b; Borrelia, Coxiella, and Rickettsia a,b	West Nile fever	[14,15]
C. vespertilionis	“Issyk-Kul” virusa	unknown	[16]
Orithodorous
O. tartakovskyi	B. latyshevyi	Tick-borne relapsing fever	[17]
O. tholozani	B. persiea	Tick-borne relapsing fever	[17]

aThese pathogenic microorganisms have been recorded outside China.

bThese pathogenic species have been detected within ticks or have been shown to be transmitted by ticks under controlled experimental conditions.

Ixodid ticks in China, their roles as vectors, and associated tick-borne pathogens

There are 104 species of ixodid ticks in China in seven genera: Amblyomma (eight species), Anomalohimalaya (two species), Dermacentor (twelve species), Haemaphysalis (forty four species), Hyalomma (six species), Ixodes (twenty four species), and Rhipicephalus (eight species) [4]. Of these, 52 species from six genera have been shown to carry or transmit pathogenic microorganisms: Ixodes (seven species), Amblyomma (three species), Dermacentor (nine species), Haemaphysalis (twenty one species), Hyalomma (five species), and Rhipicephalus (seven species) (Table 2) [18-114]. Of these 52 species, 32 occur in China (Table 2). Tick-borne spotted fever is the most commonly detected disease, carried by at least 27 tick species. Lyme disease and human granulocytic anaplasmosis are the second and third most widespread tick-borne diseases, transmitted by at least 13 and 10 tick species, respectively (Table 2). Eight tick species are vectors for human granulocytic ehrlichiosis, seven tick species carry tick-borne encephalitis and babesiosis, and six species transmit hemorrhagic fever. The ixodid ticks that act as vectors of Babesia are usually coinfected with more than one Babesia species. These ticks include I. persulcatus, D. nuttalli, Rh. microplus, and Rh. haemaphysaloides, which are often infected by Babesia bigemina and Ba. bovis (Table 2).

Table 2

Tick-borne pathogens and the role of ixodid ticks as vectors within China

Tick species	Pathogens	Diseases	References
Ixodes
I. persulcatus	B. burgdorferi; human granulocytc Ehrlichia (HGE)b; Spotted Fever Group Rickettsia (SFGR) b; Ehrlichia b; Anaplasma phagocytophila b; tick-borne Encephalitis virus (TBEV); Babesia bigemina, Ba. bovis	Lyme disease; Ehrlichiosis; spotted fever; human granulocytic anaplasmosis; babesiosis	[18-25]
I. kazakstani	B. burgdorferi a	Lyme disease	[26]
I. nipponensis	B. afzelii a,b; TBEV a,b	Lyme disease; tick-borne encephalitis;	[27,28]
I. ovatus	Ehrlichia a,b; TBEV; R. japonica a,b	Ehrlichiosis; tick-borne encephalitis; Oriental spotted fever	[29-31]
I. granulates	B. burgdorferi	Lyme disease	[32]
I. acutitarsus	B. burgdorferi	Lyme disease	[33]
I. sinensis	R. monacensis b; Ehrlichia, Bartonella, and Borrelia b; B. garinii b	Lyme disease; Mediterranean Spotted Fever	[34]
Amblyomma
Am. geoemydae	Reptile-associated Borrelia spp. a,b;relapsing fever Borrelia sp. a,b	Relapsing fever	[35]
Am. helvolum	SFGR a,b; Rickettsia sp. a,b	Spotted fever	[36,37]
Am. testudinarium	R. tamurae sp. nov.a,b; Ehrlichia chaffeensis b;	Human monocytic ehrlichiosis	[38,39]
Haemaphysalis
H. longicornis	New bunyavirusb; B. burodorferi; A. phagocytophilum b; SFGRa,b; Babesia spa; Huaiyangshan virusb; Borrelia, Bartonella, Anaplasma, and Ehrlichia b; Theileria uilenbergi b;	Severe fever with thrombocytopenia syndrome; Lyme disease; human granulocytic anaplasmosis; spotted fever; babesiosis; Huaiyangshan hemorrhagic fever	[40-47]
H. concinna	B. garinii b; HGEb; SFGRb; TBEVb	Human granulocytic Ehrlichiosis	[23,48-50]
H. punctata	B. burgdorferi sensu strictob; Ba. major and T. orientalis; Crimean-Congo hemorrhagic fever virusa; Rickettsia a,b; R. aeschlimannii a,b; An. phagocytophilum b; Flavivirus b	Lyme disease; Babesiosis; tick-borne encephalitis; Crimean-Congo hemorrhagic fever	[3,51-55]
H. cornigera	R. heilongjiangensis a,b	Spotted fever	[31]
H. erinacei	SFGRa,b	Spotted fever	[56]
H. flava	Ehrlichia b; R. japonica a,b	EhIichiosis; Japanese Spotted fever	[57,58]
H. formosensis	R. asiatica sp. nov.a,b; Kyasanur Forest disease virusa,b; R. japonica a,b; An. phagocytophilum a,b	Spotted fever; Kyasanur Forest disease;	[31,59-61]
H. hystricis	An. phagocytophilum b; R. japonica a,b	Human granulocytic anaplasmosis; Japanese Spotted fever	[31,62]
H. japonica	B. garinii b; TBEVa,b	Lyme disease; tick-borne encephalitis	[63,64]
H. kitaokai	SFGRa,b	Spotted Fever	[65]
H. lagrangei	Anaplasma spp.a,b; Rickettsia a,b;	Human granulocytic ehrlichiosis; Rickettsioses	[66,67]
H. bispinosa	B. burodorferi; T. sergenti; Ba. bigemina	Lyme disease; Piroplasmosis	[32,68,69]
H. megaspinosa	A. bovis and An. phagocytophilum a,b; SFGRa,b	Human granulocytic anaplasmosis; spotted fever;	[65,70]
H. ornithophila	SFGRa,b	Spotted Fever	[71]
H. phasiana	TBEVa,b	Tick-borne encephalitis	[64]
H. qinghaiensis	T. uilenbergi b; An. phagocytophilum b; Theileria spp.;	Human granulocytic anaplasmosis; theileriosis	[72-74]
H. spinigera	Flavivirusa,b	Kyasanur forest disease	[3]
H. tibetensis	GRD spirochetes	Unknown	[75]
H. wellingtoni	Kyasanur forest disease virusa,b; Eubacterium sp. strain Hw124 and Eubacterium sp. strain Hw191a,b	Kyasanur forest disease;	[66,76]
H. campanulata	Coxiella burneti b	Q fever	[77]
H. yeni	R. sibirica b	North-Asia Spotted fever	[78]
Hyalomma
Hy. anatolicum	Trypanosoma theileri-like flagellatesa,b; Crimean-Congo haemorrhagic fever virusa,b; T. annulata	Crimean-Congo haemorrhagic fever	[79-81]
Hy. asiaticum	Hemorrhagic fever virusb; R. mongolotimonae b; T. annulata; Rickettsiale b	Hemorrhagic fever; theileriosis	[81-84]
Hy. scupense	T. annulata	Theileriosis	[81]
Hy. rufipes	T. annulata a,b	Theileriosis	[85]
Hy. dromedarii	Kadam virusa,b	Unknown	[86]
Dermacentor
D. nuttalli	B. burgdorferi b; SFGRb; HGE; Ba. caballi and Ba. equi a,b; Rickettsiaes	Lyme disease; North Asia Spotted Fever; human granulocytic ehrlichiosis; babesiosis	[23,84,87-89]
D. reticulatus	R. helvetica a,b; R. slovaca a,b; An. phagocytophila a,b; Babesia a,b	Unexplained febrile illness; Spotless rickettsiosis; human granulocytic anaplasmosis; babesiosis	[90-93]
D. silvarum	HGEb; E. chaffeensis b; TBEV; R. raoultii sp. nov.b; R. heilongjiangensis a,b, R. sibirica b	Human granulocytic ehrlichiosis; Encephalitis; Spotted Fever;	[23,94-98]
D. auratus	SFGRb	Spotted Fever	[99]
D. everestianus	An. ovis b; R. raoultii–like bacteriab; Bacillus tularensis b	Spotted Fever; Tularemia	[100-102]
D. marginatus	B. burgdorferi b; R. slovaca a,b	Lyme disease; tick-borne lymphadenopathy	[51,103]
D. niveus	SFGRa,b; An. ovis b; Bacillus tularensis b	Spotted Fever; Tularemia	[100,102,104]
D. sinicus	SFGRb	Spotted fever	[105]
D. taiwanensis	R. japonica a,b;	Japanese Spotted fever	[106]
Rhipicephalus
Rh. microplus	A. marginale b, Ba. bigemina, Ba. bovis, T. equi; E. chaffeensis a,b; TBEVb; C. burnetii b	Babesiosis; theileriosis; Encephalitis; Q fever	[2,107,108]
Rh. bursa	An. marginale b, An. ovis b, An. phagocytophilum b, Ba. bigemina b, Ba. ovis b, Bhanja virusa,b, Crimean-Congo hemorrhagic fever virusa,b	Human granulocytic anaplasmosis; babesiosis; Crimean-Congo hemorrhagic fever	[3,109]
Rh. pumilio	SFGRa,b; R. conorii b; An. phagocytophilum b	Spotted fever; human granulocytic anaplasmosis	[110,111]
Rh. rossicus	West Nile virusa,b	West Nile fever	[112]
Rh. sanguineus	Ba. vogeli b, E. canis b, R. conorii a,b, R. massiliae a,b, R. rickettsii a,b	Babesiosis; Mediterranean spotted fever	[3,113]
Rh. turanicus	R. conorii a,b, R. massiliae a,b	Mediterranean spotted fever	[114]
Rh. haemaphysaloides	SFGRb; Ba. bigemina b, B. bovis b	Spotted fever; Babesiosis	[109]

aThese pathogenic microorganisms have been recorded outside China.

bThese pathogenic species have been detected within ticks or have been shown to be transmitted by ticks under controlled experimental conditions.

Genus Ixodes

Ixodes persulcatus is undoubtedly the most notorious tick within China, and is known to carry a wide range of microorganisms, including Borrelia, Ehrlichia, Rickettsia, Anaplasma, and Babesia [18-25]. Lyme disease is mainly transmitted by Ixodes ticks, and Borrelia spp. have been isolated from or detected in I. persulcatus, I. kazakstani, I. nipponensis, I. granulates, I. acutitarsus, and I. sinesis in China [18,26,27,32-34]. Tick-borne encephalitis virus is carried by I. persulcatus, I. nipponensis, and I. ovatus [24,28,30], whereas spotted fever can only be transmitted by I. persulcatus and I. ovatus [20,31].

Among these Ixodes species, only I. kazakstani and I. nipponensis have not yet been shown to carry Lyme disease in China, because B. burgdorferi has not been found in I. kazakstani collected in China [26], and B. afzelii has not been detected in I. nipponensis distributed in China [28]. Although their pathogens have not been confirmed in China, the vector roles of these ticks are widely recognized [26,28]. Tick-borne encephalitis virus has not been found in I. kazakstani in China [27], whereas Ehrlichia and R. japonica have only been found in the species I. ovatus, distributed outside China [29,31].

Genus Amblyomma (Am.)

Amblyomma geoemydae [35], Am. helvolum [36,37], and Am. testudinarium [38,39], collected from Japan, Thailand, and China, are known to carry pathogen DNA from Borrelia, Rickettsia, and Ehrlichia, respectively. However, although all these species are found in China, E. chaffeensis, detected in Am. testudinarium, is the only bacterial species that has been found in specimens collected within China [38].

Genus Haemaphysalis

The majority of ixodid ticks found in China belong to the genus Haemaphysalis. Globally, 21 of the 44 species found within China are known to be associated with pathogens. Of these 21 species, 11 (H. longicornis, H. concinna, H. punctata, H. flava, H. hystricis, H. japonica, H. bispinosa, H. qinghaiensis, H. tibetensis, H. campanulata, and H. yeni) have been confirmed as pathogen vectors in China (Table 2) [40-78]. The most commonly detected diseases vectored by this genus of ticks are spotted fever and human granulocytic anaplasmosis, which are transmitted by 11 and six species, respectively. Borrelia is carried by at least five species of this genus, and Babesia by at least four species (Table 2).

The ticks H. longicornis, H. punctata, and H. concinna support the greatest diversity of pathogenic microorganisms, with H. longicornis the major vector of B. burgdorferi, Theileria spp., Coxiella burnetti, Babesia spp., Anaplasma phagocytophilum, Ehrlichia, Bartonella, spotted-fever-group rickettsiae, Huaiyangshan virus, and the recently identified New bunyavirus (Table 2), which has caused many deaths in China, Japan, and Korea [40-47]. Haemaphysalis concinna is mainly distributed in northern China, where multiple outbreaks of H. concinna-borne disease have been reported since the early 20th century. These outbreaks have been attributed to a diverse array of pathogens, including B. garinii, human granulocytic Ehrlichia, spotted-fever-group Rickettsiae, and encephalitis viruses [23,48-50]. Haemaphysalis punctata transmits B. burgdorferi sensu stricto, Ba. major, T. orientalis, Crimean–Congo hemorrhagic fever virus, Rickettsia, R. aeschlimannii, An. phagocytophilum, and Flavivirus, resulting in diseases such as Lyme disease, babesiosis, tick-borne encephalitis, and Crimean-Congo hemorrhagic fever [3,51-55]. Haemaphysalis formosensis has been shown to carry pathogen DNA from a number of bacterial species, including R. asiatica sp. nov., Kyasanur Forest disease virus, R. japonica, and An. phagocytophilum, but these pathogens have not yet been detected in this tick species within China. Among the pathogenic microorganisms transmitted by Haemaphysalis species, most have been characterized with molecular techniques, and some species have been shown to transmit particular pathogens under controlled experimental conditions (Table 2).

Genus Hyalomma (Hy.)

Five species of Hyalomma are known to harbor pathogenic microorganisms (Table 2) [79-86], and three have been confirmed as vectors within China: Hy. anatolicum, Hy. asiaticum, and Hy. scupense. Hyalomma anatolicum and Hy. asiaticum carry the greatest diversity of pathogens, and each transmits at least three pathogens. Theileria annulata is the most common pathogenic microorganism, and is transmitted by four of the five Hyalomma vector ticks (Hy. anatolicum, Hy. asiaticum, Hy. scupense, and Hy. rufipes) [79,85]. Trypanosoma theileri-like flagellates and Crimean-Congo hemorrhagic fever virus have been detected in Hy. anatolicum outside China, whereas T. annulata was characterized from Hy. anatolicum within China [79-81]. Hemorrhagic fever virus and R. mongolotimonae were detected in Hy. asiaticum in north China [81-83], and Hy. asiaticum is the only tick species that can transmit Rickettsiae [84]. Hyalomma dromedarii has been shown to transmit Kadam virus outside China, although the resulting symptoms are still unknown [86].

Genus Dermacentor

Nine of the 12 species of Dermacentor found within China can transmit pathogens, and seven of these species (D. nuttali, D. silvarum, D. auratus, D. everestianus, D. marginatus, D. niveus, and D. sinicus) are of epidemiological importance in China (Table 2) [23,84-106]. The widely distributed D. nuttalli, D. reticulates, and D. silvarum carry the largest numbers of different pathogenic microorganisms, and Rickettsiae are the most commonly found bacteria in this genus (Table 2). The causative agent of human granulocytic ehrlichia has been detected in D. silvarum and D. nuttalli within China [23], and Babesia is commonly found in D. nuttalli and D. reticulatus outside China [89]. Borrelia burgdorferi has been found in D. marginatus [51] and D. nuttalli within China [87]; An. ovis and Bacillus tularensis are most commonly found in D. everestianus [100] and D. niveus within China [104]; and An. phagocytophila is specifically detected in D. reticulatus outside China [92].

Genus Rhipicephalus (Rh.)

In the genus Rhipicephalus, seven tick species are known to harbor pathogenic microorganisms, and five of these species (Rh. microplus, Rh. bursa, Rh. pumilio, Rh. sanguineus, and Rh. haemaphysaloides) are confirmed vectors in China (Table 2) [107-114]. Rhipicephalus microplus and Rh. bursa carry the largest numbers of different pathogens in this genus. Ehrlichia chaffeensis, Bhanja virus, and Crimean–Congo hemorrhagic fever virus have not yet been detected in Rh. microplus or Rh. bursa within China. Babesia is the most common microorganism transmitted by Rh. microplus [2], Rh. bursa, Rh. sanguineus [3], and Rh. haemaphysaloides [109], and various Rickettsia species have been found in Rh. pumilio [110], Rh. sanguineus, Rh. haemaphysaloides, and Rh. turanicus. West Nile virus and Crimean–Congo hemorrhagic fever virus have been characterized solely in Rh. rossicus [112] and Rh. bursa [109], respectively, whereas Ehrlichia has been found in both Rh. microplus [107] and Rh. sanguineus [113]. Anaplasma marginale, An. ovis, and An. phagocytophilum can be acquired by Rh. bursa [3,113]. Among these bacterial species, no spotted-fever-group Rickettsia has been detected in Rh. pumilio collected in China; West Nile virus has not been characterized in Rh. rossicus within China; R. conori, R. massiliae, and R. rickettsii have not been detected in Rh. sanguineus in China; and R. massiliae has not been detected in Rh. turanicus within China.

Conclusion

Of the estimated 117 species of ticks in China, 36 have been confirmed to carry or transmit one or more pathogens, and 24 additional species are known to be pathogenic vectors in other countries. Furthermore, 38 species have been shown to carry multiple pathogens, indicating the major roles they play in the spread and transmission of these pathogens. Therefore, the number of pathogens and the vector potential of ticks may still be underestimated, because of the complex distributions and the great diversity of tick species in diverse ecological habitats in China. However, such knowledge will provide clues to the further identification of tick-associated pathogens, especially in epidemic areas with multiple tick species. Much more work is required to better distinguish between ticks that carry potential pathogens and those that are competent to transmit pathogens to a host. Targeted prevention methods will then be more effective in controlling tick-borne diseases.