Abstract
PCR-based single-strand conformation polymorphism (SSCP) analyses combined with DNA sequencing of the prokaryotic 16S ribosomal (r) RNA gene encompassing the hypervariable V4 region was used to determine the bacterial composition of Rocky Mountain wood ticks (Dermacentor andersoni) attached to Richardson’s ground squirrels (Urocitellus richardsonii) and questing on vegetation in southern Saskatchewan, Canada. The bacteria present in questing adult ticks from Saskatchewan Landing Provincial Park included Rickettsia peacockii, a Francisella-like endosymbiont (FLE) and an Arsenophonus-like endosymbiont. Bacteria in the adult and nymphal ticks attached to U. richardsonii collected from Beechy included R. peacockii, a FLE, and several other genera (e.g., Ralstonia, Sphingobium, Comamonas and Pseudomonas). The bacteria detected in D. andersoni in the present study are consistent with the findings of other studies that have characterized the microbiome of this tick species in the USA using next generation sequencing. This result demonstrates that the SSCP-based approach used in this study is cost- and time-effective for examining bacterial composition in ticks.
Similar content being viewed by others
References
Andreotti R, Perez de Leon AA, Dowd SE, Guerrero FD, Bendele KG, Scoles GA (2011) Assessment of bacterial diversity in the cattle tick Rhipicephalus (Boophilus) microplus through tag-encoded pyrosequencing. BMC Microbiol 11:6
Anstead CA, Chilton NB (2011) Ticks feeding on northern pocket gophers (Thomomys talpoides) in central Saskatchewan and the unexpected detection of Ixodes scapularis larvae. J Vector Ecol 36:355–360
Anstead CA, Hwang YT, Chilton NB (2013) Ticks (Acari: Ixodidae) on small mammals in Kootenay National Park, British Columbia, Canada. J Med Entomol 50:1208–1214
Anstead CA, Wallace S, Chilton NB (2014) Mutation scanning-based identification of larval and nymphal ticks (Acari: Ixodidae) from Richardson’s ground squirrels (Spermophilus richardsonii). Mol Cell Probes 28:6–9
Burgdorfer W (1975) A review of Rocky Mountain spotted fever (tick-borne typhus), its agent, and its tick vectors in the United States. J Med Entomol 12:269–278
Burgdorfer W, Hayes SF, Mavros AJ (1981) Nonpathogenic rickettsiae in Dermacentor andersoni: a limiting factor for the distribution of Rickettsia rickettsii. In: Burgdorfer W, Anacker RL (eds) Rickettsiae and rickettsial diseases. Academic Press, New York, pp 585–594
Busse H-J, Denner EBM, Lubitz W (1996) Classification and identification of bacteria: current approaches to an old problem. Overview of methods used in bacterial systematics. J Biotechnol 47:3–38
Carpi G, Cagnacci F, Wittekindt NE, Zhao F, Qi J, Tomsho LP, Drautz DI, Rizzoli A, Schuster SC (2011) Metagenomic profile of the bacterial communities associated with Ixodes ricinus ticks. PLoS ONE 6:e35604
Chilton NB, Dergousoff SJ, Lysyk TJ (2018) Prevalence of Anaplasma bovis in Canadian populations of the Rocky Mountain wood tick, Dermacentor andersoni. Ticks Tick Borne Dis 9:1528–1531
Chow KW, Weese JS, Rousseau J, Jardine CM (2018) Microbiota of field-collected Ixodes scapularis and Dermacentor variabilis from eastern and southern Ontario, Canada. Ticks Tick Borne Dis 9:235–244
Clarridge JE III (2004) Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases. Clin Microbiol Rev 17:840–862
Clay K, Klyachko N, Grindle D, Civitello D, Oleske D, Fuqua C (2008) Microbial communities and interactions in the lone star tick, Amblyomma americanum. Mol Ecol 17:4371–4381
Clayton KA, Gall CA, Mason KL, Scoles GA, Brayton KA (2015) The characterization and manipulation of the bacterial microbiome of the Rocky Mountain wood tick, Dermacentor andersoni. Parasite Vectors 8:632
Dergousoff SJ, Chilton NB (2007) Differentiation of three species of ixodid tick, Dermacentor andersoni, D. variabilis and D. albipictus, by PCR-based approaches using markers in ribosomal DNA. Mol Cell Probes 21:343–348
Dergousoff SJ, Chilton NB (2010) Detection of a new Arsenophonus-type bacterium in Canadian populations of the Rocky Mountain wood tick, Dermacentor andersoni. Exp Appl Acarol 52:85–91
Dergousoff SJ, Chilton NB (2011) Novel genotypes of Anaplasma bovis, “Candidatus Midichloria” sp. and Ignatzschineria sp. in the Rocky Mountain wood tick, Dermacentor andersoni. Vet Microbiol 150:100–106
Dergousoff SJ, Chilton NB (2012) Association of different genetic types of Francisella-like organisms with the Rocky Mountain wood tick (Dermacentor andersoni) and the American dog tick (Dermacentor variabilis) in localities near their northern distributional limits. Appl Environ Microbiol 78:965–971
Dergousoff SJ, Chilton NB (2016) Ticks on small mammals at two localities in southern Saskatchewan, Canada and the detection of Rickettsia peacockii (Rickettsiaceae) in Dermacentor andersoni (Acari: Ixodidae) nymphs. Can Entomol 148:43–51
Dergousoff SJ, Gajadhar AJA, Chilton NB (2009) Prevalence of Rickettsia species in Canadian populations of Dermacentor andersoni and D. variabilis. Appl Environ Microbiol 75:1786–1789
Dergousoff SJ, Galloway TD, Lindsay LR, Curry P, Chilton NB (2013) Range expansion of Dermacentor variabilis and D. andersoni (Acari: Ixodidae) near their northern distributional limits. J Med Entomol 50:510–520
Foley JE, Nieto NC (2010) Tularemia. Vet Microbiol 140:332–338
Gage KL, Schrumpf ME, Burgdorfer W, Schwan TG (1994) DNA typing of rickettsiae in naturally infected ticks using a polymerase chain reaction/restriction fragment length polymorphism system. Am J Trop Med Hyg 50:247–260
Gall CA, Reif KE, Scoles GA, Mason KL, Mousel M, Noh SM, Brayton KA (2016) The bacterial microbiome of Dermacentor andersoni ticks influences pathogen susceptibility. ISME J 10:1846–1855
Gall CA, Scoles GA, Magori K, Mason KL, Brayton KA (2017) Laboratory colonization stabilizes the naturally dynamic microbiome composition of field collected Dermacentor andersoni ticks. Microbiome 5:133
Gasser RB, Hu M, Chilton NB, Campbell BE, Jex A, Otranto D, Cafarchia C, Beveridge I, Zhu X (2006) Single-strand conformation polymorphism (SSCP) for the analysis of genetic variation. Nat Protoc 1:3121–3128
Gregson JD (1956) The ixodoidea of Canada (Pub #930). Science Service, Entomology Division, Canada Department of Agriculture, Ottawa, p 92
Humphreys FA, Campbell AG (1947) Plague, Rocky Mountain spotted fever, and tularaemia surveys in Canada. Can J Public Health 38:124–130
James AM, Freier JE, Keirans JE, Durden LA, Mertins JW, Schlater JL (2006) Distribution, seasonality, and hosts of the Rocky Mountain wood tick in the United States. J Med Entomol 43:17–24
Kocan KM, de la Fuente J, Blouin EF, Coetzee JF, Ewing SA (2010) The natural history of Anplasma marginale. Vet Parasitol 167:95–107
Krakowetz CN, Dergousoff SJ, Chilton NB (2010) Genetic variation in the mitochondrial 16S rDNA of the American dog tick, Dermacentor variabilis (Acari: Ixodidae). J Vector Ecol 35:163–173
Krakowetz CN, Lindsay LR, Chilton NB (2011) Genetic diversity in Ixodes scapularis (Acari: Ixodidae) from six established populations in Canada. Ticks Tick Borne Dis 2:143–150
Lalzar I, Harrus S, Mumcuoglu KY, Gottlieb Y (2012) Composition and seasonal variation of Rhipicephalus turanicus and Rhipicephalus sanguineus bacterial communities. Appl Environ Microbiol 78:4110–4116
Niebylski ML, Peacock MG, Fischer ER, Porcella SF, Schwan TG (1997a) Characterization of an endosymbiont infecting wood ticks, Dermacentor andersoni, as a member of the Genus Francisella. Appl Environ Microbiol 63:3933–3940
Niebylski ML, Schrumpf ME, Burgdorfer W, Fischer ER, Gage KL, Schwan TG (1997b) Rickettsia peacockii sp. nov., a new species infecting wood ticks, Dermacentor andersoni, in Western Montana. Int J Syst Bacteriol 47:446–452
Noguchi H (1926) Cultivation of Rickettsia-like microorganisms from the Rocky Mountain spotted fever tick, Dermacentor andersoni. J Exp Med 43:515–532
Patterson EI, Dergousoff SJ, Chilton NB (2009) Genetic variation in the 16S mitochondrial DNA gene of two Canadian populations of Dermacentor andersoni (Acari: Ixodidae). J Med Entomol 46:475–481
Philip RN, Casper EA, Burgdorfer W, Gerloff RK, Hughes LE, Bell EJ (1978) Serologic typing of rickettsiae of the spotted fever group by microimmunofluorescence. J Immunol 121:1961–1968
Schabereiter-Gurtner C, Lubitz W, Rölleke S (2003) Application of broad-range 16S rRNA PCR amplification and DGGE fingerprinting for detection of tick-feeding bacteria. J Microbiol Methods 52:251–260
Scoles GA (2004) Phylogenetic analysis of the Francisella-like endosymbionts of Dermacentor ticks. J Med Entomol 41:277–286
Sparagano OAE, Allsopp MTEP, Mank RA, Rijpkema SGT, Figueroa JV, Jongejan F (1999) Molecular detection of pathogen DNA in ticks (Acari: Ixodidae): a review. Exp Appl Acarol 23:929–960
Steinhaus EA (1942) The microbial flora of the Rocky Mountain wood tick, Dermacentor andersoni Stiles. J Bacteriol 44:397–404
Teng J, Lindsay R, Bartlett K, Klinkenberg B, Dibernardo Wood AH, Morshed M (2011) Prevalence of tick-borne pathogens in the South Okanagan, British Columbia: active surveillance in ticks (Dermacentor andersoni) and deer mice (Peromyscus maniculatus). BCMJ 53:122–127
Varela-Stokes AS, Park SH, Stokes JV, Gavron NA, Lee SI, Moraru GM, Ricke SC (2018) Tick microbial communities within enriched extracts of Amblyomma maculatum. Ticks Tick Borne Dis 9:798–805
Wilkinson PR (1967) The distribution of Dermacentor ticks in Canada in relation to bioclimatic zones. Can J Zool 45:517–537
Acknowledgements
Funding for this work was provided from the Natural Sciences Research Council of Canada (NSERC) and the Canadian Foundation for Innovation Fund (to NBC). SJD received funding through a NSERC Postgraduate Scholarship. CAA received funding through the Margaret MacKay Scholarship and the University of Saskatchewan.
Author information
Authors and Affiliations
Corresponding author
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Dergousoff, S.J., Anstead, C.A. & Chilton, N.B. Identification of bacteria in the Rocky Mountain wood tick, Dermacentor andersoni, using single-strand conformation polymorphism (SSCP) and DNA sequencing. Exp Appl Acarol 80, 247–256 (2020). https://doi.org/10.1007/s10493-019-00459-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10493-019-00459-0