Open Access

Novel Borrelia species detected in echidna ticks, Bothriocroton concolor, in Australia

  • Siew-May Loh1,
  • Alexander W. Gofton1,
  • Nathan Lo2,
  • Amber Gillett3,
  • Una M. Ryan1,
  • Peter J. Irwin1 and
  • Charlotte L. Oskam1Email author
Parasites & Vectors20169:339

https://doi.org/10.1186/s13071-016-1627-x

Received: 5 April 2016

Accepted: 7 June 2016

Published: 14 June 2016

Abstract

Background

To date, little has been documented about microorganisms harboured within Australian native ticks or their pathogenic potential. Recently, a Borrelia sp. related to the Relapsing Fever (RF) group was identified in a single tick removed from a wild echidna (Tachyglossus aculeatus). The present study investigated the presence of Borrelia in 97 Bothriocroton concolor ticks parasitizing echidnas in Queensland, New South Wales, and Victoria, Australia, using nested PCR with Borrelia-specific primers targeting the 16S rRNA (16S) and flaB genes.

Results

Borrelia-specific PCR assays confirmed the presence of a novel Borrelia sp. related to the RF and reptile-associated (REP) spirochaetes in 38 (39 %) B. concolor ticks. This novel Borrelia sp. was identified in 41 % of the B. concolor ticks in Queensland and New South Wales, but not in any ticks from Victoria. The resulting flaB sequences (407 bp) were 88 and 86 % similar to the flaB sequences from Borrelia turcica and Borrelia hermsii, respectively. Of the ticks confirmed as Borrelia-positive following the flaB assay, 28 were positive with the 16S assay. Phylogenetic analysis of the 16S sequences (1097 bp) suggests that these sequences belong to a novel Borrelia sp., which forms a unique monophyletic clade that is similar to, but distinct from, RF Borrelia spp. and REP-associated Borrelia spp.

Conclusions

We conclude that the novel Borrelia sp. identified in this study does not belong to the Borrelia burgdorferi (sensu lato) complex, and that the phylogenetic analysis of the partial 16S gene sequences suggests it forms a unique monophyletic cluster in the genus Borrelia, potentially forming a fourth major group in this genus associated with monotremes in Australia. However, a thorough molecular characterisation will be required to confirm the phylogenetic position of this unique Borrelia sp. The zoonotic potential and pathogenic consequences of this novel Borrelia sp. are unknown at the current time.

Keywords

Tick-borne disease Bothriocroton concolor Borrelia EchidnaAustralia

Background

Ticks (Acari: Ixodida) transmit the greatest diversity of zoonotic pathogens, including bacteria, protozoa and viruses, of any arthropod and are of major concern to the health and wellbeing of humans, wildlife, livestock and companion animals [1]. Globally, ticks are associated with the transmission of many bacterial pathogens; those of most concern belong predominantly to the genera Anaplasma [2], Borrelia [3], Ehrlichia [4], Francisella [5] and Rickettsia [6]. In Australia, enzootic ticks that parasitise humans and companion animals also serve as hosts for zoonotic pathogens [7].

In Australia, 70 native and introduced tick species have been described, comprising 56 hard ticks (family Ixodidae: genera Amblyomma, Bothriocroton (formerly Aponomma), Haemaphysalis, Ixodes, and Rhipicephalus,) and 14 soft ticks (family Argasidae: genera Argas and Ornithodoros) [8]. Only a few of these ticks are known to transmit pathogens associated with human tick-borne diseases (TBD); currently there are three zoonotic TBDs recognised in Australia associated with native tick species. For example, Rickettsia australis and Rickettsia honei, the causative agents of Queensland tick typhus and Flinders Island Spotted Fever, respectively, are associated with Ixodes holocyclus (Australian paralysis tick), Bothriocroton hydrosauri (reptile tick), and Ixodes tasmani (marsupial tick) [912]. In addition, Coxiella burnetii, the causative agent of Q fever, has been detected in Haemaphysalis humerosa, the bandicoot tick [13, 14], and Amblyomma triguttatum, the ornate kangaroo tick [15]. These ticks are thought to play a role in the life-cycle of C. burnetii and this pathogen has been detected in a number of wild animals [16, 17]. Although wildlife are often suggested to be reservoirs for such pathogens in Australia and abroad [16, 18], the importance of wildlife ticks in the ecology of these pathogens is often overlooked or neglected.

Spirochaetes in the genus Borrelia are transmitted by arthropods and are classified traditionally into two groups: Lyme Borreliosis (LB) Borrelia burgdorferi (sensu lato) (s.l.), transmitted by several species of ixodid ticks, and Relapsing Fever (RF) Borrelia, transmitted primarily by argasid ticks (tick-borne RF, TBRF) [19], a few species by ixodid ticks [2023], and also by lice [24]. The LB group is the most significant from a human health perspective [25]. The LB group consists of 18 species, of which the principal LB-causing agents include Borrelia afzelii, Borrelia burgdorferi (sensu stricto) (s.s.), and Borrelia garinii, which are transmitted by the ticks Ixodes pacificus and Ixodes scapularis in the United States, Ixodes ricinus in Europe, and Ixodes persulcatus in Europe and Asia [25].

The TBRF Borrelia group is conventionally divided geographically into ‘Old World’ RF organisms such as Borrelia crocidurae, Borrelia duttonii and Borrelia hispanica; and ‘New World’ RF species such as Borrelia hermsii, Borrelia parkeri and Borrelia turicatae [26, 27]. Although RF Borrelia are predominantly associated with soft ticks, well-known examples of hard ticks-associated TBRF Borrelia include: Borrelia miyamotoi, isolated from I. persulcatus and I. ricinus in Europe and Asia [2830] and I. scapularis and I. pacificus in north-eastern and western United States, respectively [31, 32]; Borrelia lonestari in Amblyomma americanum [33]; ‘Candidatus Borrelia texasensis’ in Dermacentor variabilis in the southern states of North America [34]; and Borrelia theileri in Rhipicephalus (Boophilus) microplus [21]. While also recorded in Europe, Asia and North America [3537], TBRF is a common bacterial infection in several regions in Africa resulting in febrile illness and spirochaetaemia [3840].

In 2003, a novel Borrelia sp. was isolated from a hard tick, Hyalomma aegyptium, removed from a tortoise in Istanbul, Turkey [41]. This species was later named Borrelia turcica [41, 42] and is genetically distinct from the LB and RF spirochaetes. Further studies have since supported a third major Borrelia group classification, designated the reptile-associated (REP) Borrelia sp. group [43].

In Australia, three borreliae have been reported: B. theileri, the causative agent of bovine spirochaetosis worldwide, transmitted by the cattle tick, R. (Boophilus) australis [44, 45]; Borrelia anserina associated with poultry and transmitted by the soft tick, Argas persicus [7, 46, 47]; and Borrelia queenslandica from long-haired rats, Rattus villosissimus, in north-west Queensland, which at the time could not be detected within the proposed tick vector, Ornithodoros gurneyi [48]. Spirochaetes have also been observed within blood films of bandicoots, cattle, kangaroo and rodents [49] and in I. holocyclus and Haemaphysalis spp. ticks, collected from companion animals and livestock [50].

Echidnas, also known as spiny anteaters, are egg-laying mammals classified under the order Monotremata and belonging in the family Tachyglossidae [51]. The short-beaked echidna (T. aculeatus) is found in Australia and New Guinea, and is comprised of five subspecies (ssp.): T. a. acanthion, T. a. aculeatus, T. a. multiaculeatus, and T. a. setosus, can be found exclusively in Australia, while T. a. lawesii is found in New Guinea [51]. In a recent molecular survey of bacteria associated with native Australian human-biting ticks, a novel Borrelia sp. related to the RF group was identified in a single I. holocyclus tick removed from an echidna host [52]. This finding prompted the current investigation to further assess the occurrence and phylogenetic position of Borrelia sp. in ticks collected from echidnas in three regions of Australia and to provide greater insight into its distribution in Australia.

Methods

Tick sample collection and identification

A total of 97 ticks were collected from 22 echidnas (T. aculeatus ssp.) by veterinarians at the Australian Zoo Wildlife Hospital in Beerwah, Queensland (n = 81), wildlife carers at the Wild Days Wildlife shelter in Narre Warren, Victoria (n = 4), and through public submission from Wagga Wagga, western New South Wales (n = 12). Ticks were preserved in 70 % ethanol immediately after removal and sent to Murdoch University for species identification and molecular analyses. Twenty-six male and 71 female B. concolor ticks were identified based on morphological assessment according to the standard keys for identifying Australian ticks [53].

DNA extraction

Prior to DNA extraction, ticks were surface-sterilised with 10 % sodium hypochlorite and washed with sterile and DNA-free water, and 70 % ethanol. The extractions were carried out as described by Gofton et al. [52]. Negative controls were treated in an identical manner. Borrelia afzelii and B. burgdorferi (s.s.) DNA previously extracted from questing nymphal I. ricinus ticks (LN1, LN6, LN7 and LN9) from Leipzig, Germany [52], were used as positive controls in all PCR assays. DNA from one Borrelia sp.-infected female I. holocyclus tick (NL230) described in Gofton et al. [52], collected from an echidna host in New South Wales was also reanalysed in the present study.

Borrelia-specific PCR and sequencing

To determine the presence of Borrelia sp. within the 97 B. concolor ticks, and one I. holocyclus tick, DNA extractions were subjected to two Borrelia genus-specific PCR assays. Borrelia-specific nested-PCR assays were conducted targeting the 16S rRNA (16S) and flaB genes (Table 1). Each 25 μl PCR reaction contained 1× PerfectTaq buffer, 2.5 mM MgCl2, 1 mM dNTPs, 400 nM of each primer, 1.25 U PerfectTaq polymerase, and 2 μl undiluted DNA. Both the primary and nested Borrelia 16S PCR assays were performed with the following thermal conditions: initial denaturation at 95 °C for 5 min, 35 cycles of denaturation at 95 °C for 30 s, annealing at 51 °C for 40 s, and extension at 72 °C for 2 min, and a final extension at 72 °C for 5 min. The flaB PCR assays were performed with an initial denaturation at 95 °C for 5 min, 35 cycles of denaturation at 95 °C for 30 s, annealing at 52 °C (primary) or 55 °C (nested) for 30 s, and extension at 72 °C for 30 s, and a final extension at 72 °C for 5 min. No-template controls and positive controls were included in all PCR assays.
Table 1

Primers used for Borrelia-specific 16S rRNA and flaB genes amplification in this study, including primer sequences, annealing temperature and expected product size

Gene

Primer

Sequence (5' – 3')

Annealing temperature

Expected product size (bp)

Reference

16S

External

    
 

Bor-16 F

TGCGTCTTAAGCATGCAAGT

   
 

Bor-1360R

GTACAAGGCCCGAGAACGTA

51 °C

1,344

This study

 

Internal

    
 

Bor-27 F

CATGCAAGTCAAACGGAATG

   
 

Bor-1232R

ACTGTTTCGCTTCGCTTTGT

51 °C

1,205

This study

flaB

External

    
 

FlaB280F

GCAGTTCARTCAGGTAACGG

   
 

FlaRL

GCAATCATAGCCATTGCAGATTGT

52 °C

645

[33, 66]

 

Internal

    
 

flaB_737F

GCATCAACTGTRGTTGTAACATTAACAGG

   
 

FlaLL

ACATATTCAGATGCAGACAGAGGT

55 °C

407

[33, 66]

Amplified PCR products were electrophoresed through 1–2 % agarose gels, stained with GelRed (Biotium), and visualised under UV light. Amplicons of expected sizes were excised from the gel and purified with the Wizard® SV Gel and PCR Clean-Up System (Promega Madison, WI, USA,), according to the manufacturer’s recommendations. Purified PCR products were sequenced with both forward and reverse nested PCR primers using BigDye v3.1 terminator on an ABI 373096 Capillary Sequencer (Life Technologies, USA).

Sequence analysis

Trimmed 16S sequences (1097 bp) generated in this study, together with sequences from other Borrelia spp. retrieved from GenBank, were aligned using MAFFT v7.017 [54], and the alignment was refined using MUSCLE [55]. Spirochaeta americana ASpG1 strain (AF373921) [56] was used as an outgroup. Following multiple sequence alignments, MEGA version 6 [57] was used to determine the most suitable nucleotide substitution model based on the Bayesian Information Criterion (BIC). General time reversible (GTR) model was selected and the Borrelia 16S phylogenetic tree was generated using FastTree 2 [58], with 20 rate categories of site and resampling 1000 times. 16S sequences generated in this study were deposited in GenBank under accessions KU954112 to KU954115, and flaB sequences were deposited in GenBank under the accession KX192143 to KX192150.

Results

Molecular and phylogenetic analyses

A partial fragment (407 bp) of the Borrelia flaB gene was successfully amplified in three male and 35 female B. concolor ticks (38/97, 39 %) and also in the single I. holocyclus tick from the previous study [52] (Table 2). All ticks (n = 4) from Victoria were negative for Borrelia spp. Bothriocroton concolor ticks that were Borrelia-positive originated from Queensland and New South Wales (38/93, 41 %) (Additional file 1: Table S1). BLAST analysis showed that, with 100 % query coverage, these flaB equences shared 87.4–88.6 % similarity with flaB sequences from B. turcica (GenBank: AB109245; AB109244; AB109241) [41, 42], and with 85.5–86.4 % similarity with B. hermsii (GenBank: AY597795; AY597798) [59]. BLAST analysis of the positive samples isolated from I. ricinus ticks revealed 100 % identity with the LB spirochaetes, with samples LN1, LN7 and LN9 identical to B. afzelii at the flaB locus (GenBank: GU826786) [60]; and LN6 identical to B. burgdorferi (GenBank: DQ016620; AF386506; AB035618) [61, 62].
Table 2

Summary of the sex and life stages of the tick specimens used in this study, including the positive controls Ixodes holocyclus and I. ricinus. The geographical regions where the ticks were collected either from a host or vegetation were recorded and the number of ticks positive for Borrelia genes is presented in terms of percentage

Ticks

Region

Host/questing

n

Gene (% of positive samples)

16S

flaB

B. concolor

     

M

QLD

Echidna

12

0

16.6

NSW

Echidna

10

10

10

VIC

Echidna

4

0

0

F

QLD

Echidna

69

36.2

50.7

NSW

Echidna

2

100

100

I. holocyclus

     

F

NSW

Echidna

1

100

100

I. ricinus

     

N

Leipzig, Germany

Questing

4

100

100

Abbreviations: M male adult, F female adult, N nymph, QLD Queensland, NSW New South Wales, VIC Victoria

The Borrelia-specific 16S PCR assay was positive for one male and 27 female B. concolor ticks (28/97; 29 %), all of which were also positive at the flaB locus (Table 2). Three distinct Borrelia 16S sequences were produced from B. concolor ticks, which differed by only a single nucleotide polymorphism (SNP) (99.9 % similarity) between the isolates. Alignment with the Borrelia sequence isolated from an I. holocyclus tick, from the previous study [52], revealed that the sequences shared > 99 % identity, with one verified SNP. The three unique Borrelia 16S sequences from B. concolor ticks were putatively designated Borrelia sp. Aus A, Borrelia sp. Aus B, and Borrelia sp. Aus C, which occurred in 22, five, and one sample, respectively. The Borrelia 16S sequence from the I. holocyclus tick was putatively designated Borrelia sp. NL230.

Phylogenetic analysis indicates that Borrelia 16S sequences from this study form a unique monophyletic clade, with high confidence, and that is most similar to, but distinct from the RF and REP Borrelia groups (Fig. 1). Borrelia 16S sequences from these echidna ticks were most dissimilar to the LB group (96.1–96.7 % similarity); and most similar (98.6–98.7 %) to both Borrelia sp. tAG66M and B. hermsii DAH (Additional file 2: Table S2). Borrelia 16S sequences generated from nymphal I. ricinus ticks clustered within the B. burgdorferi (s.l.) group, with high (99.9–100 %) similarity to B. burgdorferi and B. afzelii (Fig. 1).
Fig. 1

Molecular phylogenetic analysis of 1097 bp fragment of 16S rRNA of the Borrelia sp. amplified in B. concolor ticks from echidnas. Phylogenetic distances were inferred using FastTree 2 [57], and specimens were compared with the B. burgdorferi (sensu lato) complex, the TBRF group, and the REP-associated spirochaetes. “*” represents Borrelia sequences from B. concolor ticks; “**” indicates Borrelia sequence from Ixodes holocyclus tick; and “***” represents positive controls from I. ricinus nymphs

Discussion

This study presents the first molecular characterisation in Australia of a novel Borrelia sp. identified in B. concolor (this study) and in a single I. holocyclus tick [52], with a close relationship to the RF and REP-associated Borrelia groups. The current investigation aimed to provide further evidence for Borrelia in echidna ticks, if any, and to validate the previous findings [52] with an increased sample size and by targeting the highly conserved Borrelia housekeeping genes (e.g. flaB and 16S rRNA).

The reduced sensitivity of detecting Borrelia with the 16S PCR assay, compared to the flaB PCR assay, may be explained by the reduced PCR efficiency that is typically observed when amplifying longer gene fragments. For this reason, PCR assays targeting short gene fragments, such as the flaB assay used in the present study, are recommended for sensitive detection of Borrelia spp. However, short gene fragments are often inadequate to produce meaningful phylogenetic reconstructions, and therefore must be complimented by assays that amplify longer gene sequences, albeit with less sensitivity, such as the 16S assay in the present study.

Recently, REP-associated Borrelia spp. isolated from reptile ticks, H. aegyptium, were shown to form a monophyletic clade [42], while sharing a common ancestor with the RF and LB Borrelia groups, supporting the notion of a third Borrelia group [43]. Likewise, the Borrelia sp. identified in this study also forms a unique monophyletic clade, and may form a fourth major phylogenetic Borrelia group. However, further analyses of other Borrelia housekeeping genes, such as gyrB, groEL, glpQ genes, and 23S-16S intergenic spacer region [43], and morphological characterisation are necessary for a complete description of this novel Borrelia sp.

The current investigation has provided new records of a Borrelia sp. found in a native Australian hard tick, B. concolor [44, 45, 52]. Currently, the degree of similarity and differences between the Borrelia sp. characterised in the present study and B. queenslandica, identified in long-haired rats, R. villosissimus [48], is unknown as molecular data does not exist for this species.

All ticks removed from echidna hosts for the present study were B. concolor, a known specialist tick species usually restricted to parasitising echidnas (family Tachyglossidae) [53, 63]; however, kangaroos (Macropus fuliginosus fuliginosus) were recently recorded as a new host of B. concolor on Kangaroo Island, South Australia [64]. Bothriocroton concolor has a relatively wide distribution including both coastal and sub-coastal regions of Queensland and New South Wales, as well as inland New South Wales; whereas the distribution of I. holocyclus (the host of the first isolate reported by Gofton et al. [52]) is mainly restricted to coastal regions of the eastern Australia [53]. It has been reported that echidnas can also host other tick species such as Amblyomma australiense, Amblyomma echidnae, Amblyomma moyi, Amblyomma papuanum, Bothriocroton tachyglossi, Bothriocroton undatum, Haemaphysalis humerosa and I. tasmani [53]. Therefore, different tick species that feed on the same host may become infected through blood meals, if the echidna serves as a bacteraemic vertebrate reservoir for this bacterium.

Generally, many tick-borne microorganisms circulate within unique sylvatic cycles in an ecosystem, and wildlife and their ticks play important roles as reservoirs and bridging vectors, respectively [65]. In Australia, wildlife and their ticks have long been considered as reservoirs for a number of tick-borne pathogens [10, 16]. In the case of the vector of Borrelia in Australia, the soft tick, O. gurneyi from long-haired rats, was proposed as a vector of B. queenslandica, however, transmission attempts were unsuccessful [48]. Here in the present study, a novel bacterium harboured within B. concolor was identified in two tick genera, suggesting that echidnas may be a potential reservoir of this bacterium. However, whether these ticks are able to acquire this bacterium from infected echidna hosts remains to be confirmed through the analysis of echidna blood samples, and until further studies are completed, the vertebrate reservoir of this spirochaete remains to be determined. Likewise, the role of B. concolor and I. holocyclus ticks as potential bridging vectors of this novel Borrelia sp. remains to be assessed via transmission studies. Despite this, the potential and importance of wildlife and their ticks acting as reservoir and vector in maintaining the persistence of this bacterium in the environment cannot be disregarded. The pathogenic consequences (if any) and potential infectivity of this Borrelia sp. to animals and humans are unknown at the present time.

Conclusions

The current study has identified a novel Borrelia sp. harboured within echidna ticks. Phylogenetic analysis of the partial 16S sequences showed that this Borrelia forms a unique monophyletic clade that is closely related to the RF and REP Borrelia groups and is most dissimilar to the LB group. Moreover, this study highlights the significance of studying Australian wildlife ticks. The presence of a novel Borrelia sp. in Australia is of significant public health importance and warrants further investigations to better understand the biology, ecology, pathogenicity (if any), and infectivity of this organism to humans, domestic animals, and wildlife.

Abbreviations

16S, 16S rRNA; BIC, bayesian information criterion; GTR, general time reversible; LB, Lyme Borreliosis; NSW, New South Wales; QLD, Queensland; REP, reptile-associated; RF, relapsing fever; s.l., sensu lato; s.s., sensu stricto; SNP, single nucleotide polymorphism; TBD, tick-borne disease; TBRF, tick-borne relapsing fever; VIC, Victoria

Declarations

Acknowledgements

The authors wish to acknowledge the assistance of Reinhard Straubinger and Martin Pfeffer for the collection of Ixodes ricinus ticks in Germany. The authors acknowledge the clinicians and volunteers as well as individuals who have participated in the collection and submission of the tick samples from the Australian Zoo Wildlife Hospital and Wild Days Wildlife shelter. Many thanks to Frances Brigg and the Western Australia State Agricultural Biotechnology Centre for Sanger Sequencing, and Kevin Stratford and the Pawsey Supercomputing Centre for technical assistance during data analysis.

Funding

This study was part-funded by the Australian Research Council (LP13010005), Bayer HealthCare (Germany) and Bayer Australia and is gratefully acknowledged.

Availability of data and materials

Nucleotide sequences reported in this study were deposited in GenBank under accessions KU954112 to KU954115 and KX192143 to KX192150.

Authors’ contributions

SL performed tick identification, DNA extractions, PCR assays, data analyses, wrote the manuscript, and participated in the design of the study. AWG designed the primers, performed PCR assays on the Borrelia sp. NL230 isolate, and contributed to data analysis and manuscript preparation. NL contributed to the tick collection and identification, DNA extraction, and preparation of manuscript. AG contributed to tick samples collection and manuscript preparation. UMR and PJI contributed to the design of the study and manuscript preparation. CLO conceived, designed, coordinated the study, and contributed to the data analyses and the preparation of the manuscript. All authors read and approved the final version of the manuscript.

Competing interests

The authors declare that they have no competing interests.

Consent for publication

Not applicable.

Ethics approval and consent to participate

This study was conducted under the compliance of the Australian Code for the Responsible Conduct of Research, 2007 and Australian Code for the Care and Use of Animals for Scientific Purposes, 2013. Tick collection was carried out opportunistically with approval from the Murdoch University Animal Ethics Committee.

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Authors’ Affiliations

(1)
Vector and Water-Borne Pathogen Research Laboratory, School of Veterinary and Life Sciences, Murdoch University
(2)
School of Biological Sciences, The University of Sydney
(3)
Australia Zoo Wildlife Hospital

References

  1. Parola P, Raoult D. Ticks and tickborne bacterial diseases in humans: An emerging infectious threat. Clin Infect Dis. 2001;32:897–928.View ArticlePubMedGoogle Scholar
  2. Stuen S, Granquist EG, Silaghi C. Anaplasma phagocytophilum-a widespread multi-host pathogen with highly adaptive strategies. Front Cell Infect Microbiol. 2013;3:31.View ArticlePubMedPubMed CentralGoogle Scholar
  3. Dantas-Torres F, Chomel BB, Otranto D. Ticks and tick-borne diseases: a One Health perspective. Trends Parasitol. 2012;28:437–46.View ArticlePubMedGoogle Scholar
  4. Yabsley MJ. Natural history of Ehrlichia chaffeensis: vertebrate hosts and tick vectors from the United States and evidence for endemic transmission in other countries. Vet Parasitol. 2010;167:136–48.View ArticlePubMedGoogle Scholar
  5. Petersen JM, Mead PS, Schriefer ME. Francisella tularensis: an arthropod-borne pathogen. Vet Res. 2009;40:7.View ArticlePubMedGoogle Scholar
  6. Parola P, Paddock CD, Raoult D. Tick-borne rickettsioses around the world: Emerging diseases challenging old concepts. Clin Microbiol Rev. 2005;18:719–56.View ArticlePubMedPubMed CentralGoogle Scholar
  7. Barker SC, Walker AR. Ticks of Australia. The species that infest domestic animals and humans. Zootaxa. 2014;3816:1–144.View ArticlePubMedGoogle Scholar
  8. Barker SC, Walker AR, Campelo D. A list of the 70 species of Australian ticks; diagnostic guides to and species accounts of Ixodes holocyclus (paralysis tick), Ixodes cornuatus (southern paralysis tick) and Rhipicephalus australis (Australian cattle tick); and consideration of the place of Australia in the evolution of ticks with comments on four controversial ideas. Int J Parasitol. 2014;44:941–53.View ArticlePubMedGoogle Scholar
  9. Sexton DJ, Dwyer B, Kemp R, Graves S. Spotted fever group rickettsial infections in Australia. Rev Infect Dis. 1991;13:876–86.View ArticlePubMedGoogle Scholar
  10. Stenos J, Graves S, Popov VL, Walker DH. Aponomma hydrosauri, the reptile-associated tick reservoir of Rickettsia honei on Flinders Island, Australia. Am J Trop Med Hyg. 2003;69:314–7.PubMedGoogle Scholar
  11. Graves S, Stenos J. Rickettsioses in Australia. Ann N Y Acad Sci. 2009;1166:151–5.View ArticlePubMedGoogle Scholar
  12. Campbell RW, Domrow R. Rickettsioses in Australia: Isolation of Rickettsia tsutsugamushi and R. australis from naturally infected arthropods. Trans Roy Soc Trop Med Hyg. 1974;68:397–402.View ArticlePubMedGoogle Scholar
  13. Bennett MD, Woolford L, Banazis MJ, O’Hara AJ, Warren KS, Nicholls PK, et al. Coxiella burnetii in western barred bandicoots (Perameles bougainville) from Bernier and Dorre Islands in Western Australia. EcoHealth. 2011;8:519–24.View ArticlePubMedGoogle Scholar
  14. Smith DJW, Derrick EH. Studies in the epidemiology of Q fever 1. The isolation of six strains of Rickettsia burneti from the tick Haemaphysalis humerosa. Aust J Exp Biol Med Sci. 1940;18:1–8.View ArticleGoogle Scholar
  15. McDiarmid L, Petney T, Dixon B, Andrews R. Range expansion of the tick Amblyomma triguttatum triguttatum, an Australian vector of Q fever. Int J Parasitol. 2000;30:791–3.View ArticlePubMedGoogle Scholar
  16. Cooper A, Stephens J, Ketheesan N, Govan B. Detection of Coxiella burnetii DNA in wildlife and ticks in northern Queensland, Australia. Vector Borne Zoonotic Dis. 2013;13:12–6.View ArticlePubMedGoogle Scholar
  17. Tozer SJ, Lambert SB, Strong CL, Field HE, Sloots TP, Nissen MD. Potential animal and environmental sources of Q fever infection for humans in Queensland. Zoonoses Public Health. 2014;61:105–12.View ArticlePubMedGoogle Scholar
  18. Banazis MJ, Bestall AS, Reid SA, Fenwick SG. A survey of Western Australian sheep, cattle and kangaroos to determine the prevalence of Coxiella burnetii. Vet Microbiol. 2010;143:337–45.View ArticlePubMedGoogle Scholar
  19. Felsenfeld O. Borreliae, human relapsing fever, and parasite-vector-host relationships. Bacteriol Rev. 1965;29:46–74.PubMedPubMed CentralGoogle Scholar
  20. Trees AJ. The transmission of Borrelia theileri by Boophilus annulatus (Says, 1821). Trop Anim Health Prod. 1978;10:93–4.View ArticlePubMedGoogle Scholar
  21. Smith RD, Miranpuri GS, Adams JH, Ahrens EH. Borrelia theileri: isolation from ticks (Boophilus microplus) and tick-borne transmission between splenectomized calves. Am J Vet Res. 1985;46:1396–8.PubMedGoogle Scholar
  22. Varela-Stokes AS. Transmission of bacterial agents from lone star ticks to white-tailed deer. J Med Entomol. 2007;44:478–83.View ArticlePubMedGoogle Scholar
  23. van Duijvendijk G, Coipan C, Wagemakers A, Fonville M, Ersöz J, Oei A, et al. Larvae of Ixodes ricinus transmit Borrelia afzelii and B. miyamotoi to vertebrate hosts. Parasit Vectors. 2016;9:97.View ArticlePubMedPubMed CentralGoogle Scholar
  24. Raoult D, Roux V. The body louse as a vector of reemerging human diseases. Clin Infect Dis. 1999;29:888–911.View ArticlePubMedGoogle Scholar
  25. Rudenko N, Golovchenko M, Grubhoffer L, Oliver Jr JH. Updates on Borrelia burgdorferi sensu lato complex with respect to public health. Ticks Tick Borne Dis. 2011;2:123–8.View ArticlePubMedPubMed CentralGoogle Scholar
  26. Fukunaga M, Okada K, Nakao M, Konishi T, Sato Y. Phylogenetic analysis of Borrelia species based on flagellin gene sequences and its application for molecular typing of Lyme disease borreliae. Int J Syst Bacteriol. 1996;46:898–905.View ArticlePubMedGoogle Scholar
  27. Toledo A, Anda P, Escudero R, Larsson C, Bergstrom S, Benach JL. Phylogenetic analysis of a virulent Borrelia species isolated from patients with relapsing fever. J Clin Microbiol. 2010;48:2484–9.View ArticlePubMedPubMed CentralGoogle Scholar
  28. Fraenkel CJ, Garpmo U, Berglund J. Determination of novel Borrelia genospecies in Swedish Ixodes ricinus ticks. J Clin Microbiol. 2002;40:3308–12.View ArticlePubMedPubMed CentralGoogle Scholar
  29. Fukunaga M, Takahashi Y, Tsuruta Y, Matsushita O, Ralph D, McClelland M, et al. Genetic and phenotypic analysis of Borrelia miyamotoi sp. nov., isolated from the ixodid tick Ixodes persulcatus, the vector for Lyme disease in Japan. Int J Syst Bacteriol. 1995;45:804–10.View ArticlePubMedGoogle Scholar
  30. Platonov AE, Karan LS, Kolyasnikova NM, Makhneva NA, Toporkova MG, Maleev VV, et al. Humans infected with relapsing fever spirochete Borrelia miyamotoi, Russia. Emerg Infect Dis. 2011;17:1816–23.View ArticlePubMedPubMed CentralGoogle Scholar
  31. Mun J, Eisen RJ, Eisen L, Lane RS. Detection of a Borrelia miyamotoi sensu lato relapsing-fever group spirochete from Ixodes pacificus in California. J Med Entomol. 2006;43:120–3.View ArticlePubMedGoogle Scholar
  32. Scoles GA, Papero M, Beati L, Fish D. A relapsing fever group spirochete transmitted by Ixodes scapularis ticks. Vector Borne Zoonotic Dis. 2001;1:21–34.View ArticlePubMedGoogle Scholar
  33. Barbour AG, Maupin GO, Teltow GJ, Carter CJ, Piesman J. Identification of an uncultivable Borrelia species in the hard tick Amblyomma americanum: possible agent of a Lyme disease-like illness. J Infect Dis. 1996;173:403–9.View ArticlePubMedGoogle Scholar
  34. Lin T, Gao L, Seyfang A, Oliver Jr JH. ‘Candidatus Borrelia texasensis’, from the American dog tick Dermacentor variabilis. Int J Syst Evol Microbiol. 2005;55:685–93.View ArticlePubMedGoogle Scholar
  35. Assous MV, Wilamowski A. Relapsing fever borreliosis in Eurasia-forgotten, but certainly not gone! Clin Microbiol Infec. 2009;15:407–14.View ArticleGoogle Scholar
  36. Boyer KM, Munford RS, Maupin GO, Pattison CP, Fox MD, Barnes AM, et al. Tick-borne relapsing fever: an interstate outbreak originating at Grand Canyon National Park. Am J Epidemiol. 1977;105:469–79.PubMedGoogle Scholar
  37. Schwan TG, Raffel SJ, Schrumpf ME, Webster LS, Marques AR, Spano R, et al. Tick-borne Relapsing Fever and Borrelia hermsii, Los Angeles County, California, USA. Emerg Infect Dis. 2009;15:1026–31.View ArticlePubMedPubMed CentralGoogle Scholar
  38. Dupont HT, La Scola B, Williams R, Raoult D. A focus of tick-borne relapsing fever in southern Zaire. Clin Infect Dis. 1997;25:139–44.View ArticlePubMedGoogle Scholar
  39. Trape JF, Diatta G, Arnathau C, Bitam I, Sarih M, Belghyti D, et al. The epidemiology and geographic distribution of relapsing fever borreliosis in West and North Africa, with a review of the Ornithodoros erraticus complex (Acari: Ixodida). PLoS ONE. 2013;8(11):e78473.Google Scholar
  40. Vial L, Diatta G, Tall A, el Ba H, Bouganali H, Durand P, et al. Incidence of tick-borne relapsing fever in west Africa: longitudinal study. Lancet. 2006;368:37–43.View ArticlePubMedGoogle Scholar
  41. Guner ES, Hashimoto N, Kadosaka T, Imai Y, Masuzawa T. A novel, fast-growing Borrelia sp. isolated from the hard tick Hyalomma aegyptium in Turkey. Microbiology. 2003;149:2539–44.View ArticlePubMedGoogle Scholar
  42. Guner ES, Watanabe M, Hashimoto N, Kadosaka T, Kawamura Y, Ezaki T, et al. Borrelia turcica sp. nov., isolated from the hard tick Hyalomma aegyptium in Turkey. Int J Syst Evol Microbiol. 2004;54:1649–52.View ArticlePubMedGoogle Scholar
  43. Takano A, Goka K, Une Y, Shimada Y, Fujita H, Shiino T, et al. Isolation and characterization of a novel Borrelia group of tick-borne borreliae from imported reptiles and their associated ticks. Environ Microbiol. 2010;12:134–46.View ArticlePubMedGoogle Scholar
  44. Callow LL, Hoyte HMD. Transmission experiments using Babesia bigemina, Theileria mutans, Borrelia sp. and the cattle tick. Aust Vet J. 1961;37:381–90.View ArticleGoogle Scholar
  45. Mulhearn CR. A note on two blood parasites of cattle (Spirochaeta theileri and Bartonella bovis) recorded for the first time in Australia. Aust Vet J. 1946;22:118.View ArticlePubMedGoogle Scholar
  46. Gorrie CJR. Vaccination against spirochetosis in fowls. Aust Vet J. 1950;26:308–15.View ArticlePubMedGoogle Scholar
  47. Petney TN, Andrews RH, McDiarmid LA, Dixon BR. Argas persicus sensu stricto does occur in Australia. Parasitol Res. 2004;93:296–9.View ArticlePubMedGoogle Scholar
  48. Carley JG, Pope JH. A new species of Borrelia (B. queenslandica) from Rattus villosissimus in Queensland. Aust J Exp Biol Med Sci. 1962;40:255–61.View ArticlePubMedGoogle Scholar
  49. Mackerras MJ. The haematozoa of Australian mammals. Aust J Zool. 1959;7:105–35.View ArticleGoogle Scholar
  50. Wills MC, Barry RD. Detecting the cause of Lyme disease in Australia. Med J Aust. 1991;155:275.PubMedGoogle Scholar
  51. Augee M, Gooden B, Musser A. Echidna: extraordinary egg-laying mammal. Collingwood: CSIRO Publishing; 2006.Google Scholar
  52. Gofton AW, Oskam CL, Lo N, Beninati T, Wei H, McCarl V, et al. Inhibition of the endosymbiont “Candidatus Midichloria mitochondrii” during 16S rRNA gene profiling reveals potential pathogens in Ixodes ticks from Australia. Parasit Vectors. 2015;8:345.View ArticlePubMedPubMed CentralGoogle Scholar
  53. Roberts FHS. Australian ticks. 2nd ed. Melbourne: CSIRO; 1970.Google Scholar
  54. Katoh K, Misawa K, Kuma K, Miyata T. MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res. 2002;30:3059–66.View ArticlePubMedPubMed CentralGoogle Scholar
  55. Edgar RC. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res. 2004;32:1792–7.View ArticlePubMedPubMed CentralGoogle Scholar
  56. Hoover RB, Pikuta EV, Bej AK, Marsic D, Whitman WB, Tang J, et al. Spirochaeta americana sp. nov., a new haloalkaliphilic, obligately anaerobic spirochaete isolated from soda Mono Lake in California. Int J Syst Evol Microbiol. 2003;53:815–21.View ArticlePubMedGoogle Scholar
  57. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: Molecular evolutionary genetic analysis version 6.0. Mol Biol Evol. 2013;30:2725–9.View ArticlePubMedPubMed CentralGoogle Scholar
  58. Price MN, Dehal PS, Arkin AP. FastTree 2-approximately maximum-likelihood trees for large alignments. PLoS ONE. 2010;5:e9490.View ArticlePubMedPubMed CentralGoogle Scholar
  59. Porcella SF, Raffel SJ, Anderson Jr DE, Gilk SD, Bono JL, Schrumpf ME, et al. Variable tick protein in two genomic groups of the relapsing fever spirochete Borrelia hermsii in western North America. Infect Immun. 2005;73:6647–58.View ArticlePubMedPubMed CentralGoogle Scholar
  60. Reye AL, Hubschen JM, Sausy A, Muller CP. Prevalence and seasonality of tick-borne pathogens in questing Ixodes ricinus ticks from Luxembourg. Appl Environ Microbiol. 2010;76:2923–31.View ArticlePubMedPubMed CentralGoogle Scholar
  61. Wodecka B. flaB gene as a molecular marker for distinct identification of Borrelia species in environmental samples by the PCR-restriction fragment length polymorphism method. Appl Environ Microbiol. 2011;77:7088–92.View ArticlePubMedPubMed CentralGoogle Scholar
  62. Quessada T, Martial-Convert F, Arnaud S, Leudet De La Vallee H, Gilot B, et al. Prevalence of Borrelia burgdorferi species and identification of Borrelia valaisiana in questing Ixodes ricinus in the Lyon region of France as determined by polymerase chain reaction-restriction fragment length polymorphism. Eur J Clin Microbiol Infect Dis. 2003;22:165–73.PubMedGoogle Scholar
  63. Belan I, Bull CM. Host-seeking behaviour by Australian ticks (Acari: Ixodidae) with differing host specificities. Exp Appl Acarol. 1995;19:221–32.View ArticlePubMedGoogle Scholar
  64. Oorebeek M, Rismiller P. Bothriocroton concolor (Acari: Ixodidae) on the Kangaroo Island kangaroo: a new host-parasite relationship. J Med Entomol. 2007;44:901–2.PubMedGoogle Scholar
  65. Hamer SA, Hickling GJ, Keith R, Sidge JL, Walker ED, Tsao JI. Associations of passerine birds, rabbits, and ticks with Borrelia miyamotoi and Borrelia andersonii in Michigan, U.S.A. Parasit Vectors. 2012;5:231.View ArticlePubMedPubMed CentralGoogle Scholar
  66. Clark KL, Leydet B, Hartman S. Lyme borreliosis in human patients in Florida and Georgia, USA. Int J Med Sci. 2013;10:915–31.View ArticlePubMedPubMed CentralGoogle Scholar

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