Molecular Detection of Anaplasma phagocytophilum, Babesia odocoilei, and Borrelia burgdorferi Sensu Lato in Ixodes scapularis Ticks Collected in Veterinary Clinics in Southern Wellington County, Ontario, Canada
Tick-borne zoonotic diseases are sinister afflictions to mankind world-wide. A total of 96 adults of the blacklegged tick, Ixodes scapularis, were collected in southern Wellington County. Using molecular analysis, three pathogens were detected, namely Borrelia burgdorferi sensu lato (s.l.), 24/96 (25%), Babesia odocoilei, 15/96 (16%), and Anaplasma phagocytophilum, 1/96 (1%). A single co-infection consisting of B. burgdorferi s.l. and A. phagocytophilum was also detected. We report the first tick-host-pathogen study in southern Wellington County. Overall, 16 established populations were discovered. If clinicians only test and treat patients for the Lyme disease bacterium, they miss 40% of the tick-borne zoonotic infections.
References
[1]
Eisen, R.J., Eisen, L. and Beard, C.B. (2016) County-Scale Distribution of Ixodes scapularis and Ixodes pacificus (Acari: Ixodidae) in the Continental United States. JournalofMedicalEntomology, 53, 349-386. https://doi.org/10.1093/jme/tjv237
[2]
Cosiquien, R.J.S., Stojiljkovic, N., Nordstrom, C.W., Amadi, E., Lutwick, L. and Dumic, I. (2023) Anaplasma phagocytophilum Encephalitis: A Case Report and Literature Review of Neurologic Manifestations of Anaplasmosis. InfectiousDiseaseReports, 15, 354-359. https://doi.org/10.3390/idr15040035
[3]
Western, K.A., Benson, G.D., Gleason, N.N., Healy, G.R. and Schultz, M.G. (1970) Babesiosis in a Massachusetts Resident. NewEnglandJournalofMedicine, 283, 854-856. https://doi.org/10.1056/nejm197010152831607
[4]
Scott, J.D., Sajid, M.S., Pascoe, E.L. and Foley, J.E. (2021) Detection of Babesia odocoilei in Humans with Babesiosis Symptoms. Diagnostics, 11, Article 947. https://doi.org/10.3390/diagnostics11060947
[5]
Scott, J.D. (2024) Human Babesiosis Caused by Babesia odocoilei: A Confirmed Zoonosis. AmericanJournalofInfectiousDiseases, 20, 50-51. https://doi.org/10.3844/ajidsp.2024.50.51
[6]
Burgdorfer, W., Barbour, A.G., Hayes, S.F., Benach, J.L., Grunwaldt, E. and Davis, J.P. (1982) Lyme Disease—A Tick-Borne Spirochetosis? Science, 216, 1317-1319. https://doi.org/10.1126/science.7043737
[7]
Sroda Agudogo, J., Febres-Cordero, D. and Collier, A.Y. (2025) A Day in the Woods in Pregnancy: Fetal and Neonatal Implications. NeoReviews, 26, e57-e61. https://doi.org/10.1542/neo.26-1-006
[8]
Pritt, B.S., Allerdice, M.E.J., Sloan, L.M., Paddock, C.D., Munderloh, U.G., Rikihisa, Y., et al. (2017) Proposal to Reclassify Ehrlichia muris as Ehrlichia muris Subsp. Muris Subsp. Nov. and Description of Ehrlichia muris Subsp. Eauclairensis Subsp. Nov., a Newly Recognized Tick-Borne Pathogen of Humans. InternationalJournalofSystematicandEvolutionaryMicrobiology, 67, 2121-2126. https://doi.org/10.1099/ijsem.0.001896
[9]
Anderson, J.F. and Armstrong, P.M. (2012) Prevalence and Genetic Characterization of Powassan Virus Strains Infecting Ixodes scapularis in Connecticut. TheAmericanSocietyofTropicalMedicineandHygiene, 87, 754-759. https://doi.org/10.4269/ajtmh.2012.12-0294
[10]
Scott, J.D. and Scott, C.M. (2024) Molecular Detection of Borrelia burgdorferi Sensu Lato, Borrelia miyamotoi, Babesia odocoilei, Babesia microti and Anaplasma phagocytophilum in Ixodes Ticks Collected across Canada. JournalofBiomedicalResearch&EnvironmentalSciences, 5, 1321-1337. https://doi.org/10.37871/jbres2020
[11]
Gray, A., Capewell, P., Zadoks, R., Taggart, M.A., French, A.S., Katzer, F., et al. (2021) Wild Deer in the United Kingdom Are a Potential Reservoir for the Livestock Parasite Babesia divergens. CurrentResearchinParasitology&Vector-BorneDiseases, 1, Article ID: 100019. https://doi.org/10.1016/j.crpvbd.2021.100019
[12]
Dreghiciu, I.C., Hoffman, D., Dumitru, S., Oprescu, I., Imre, M., Florea, T., et al. (2025) First Molecular Identification of Zoonotic Babesia odocoilei in Ticks from Romania. Microorganisms, 13, Article 1182. https://doi.org/10.3390/microorganisms13061182
[13]
Holman, P.J., Madeley, J., Craig, T.M., Allsopp, B.A., Allsopp, M.T.E.P., Petrini, K.R., et al. (2000) Antigenic, Phenotypic and Molecular Characterization Confirms Babesia odocoilei Isolated from Three Cervids. JournalofWildlifeDiseases, 36, 518-530. https://doi.org/10.7589/0090-3558-36.3.518
[14]
Jalovecka, M., Sojka, D., Ascencio, M. and Schnittger, L. (2019) Babesia Life Cycle—When Phylogeny Meets Biology. TrendsinParasitology, 35, 356-368. https://doi.org/10.1016/j.pt.2019.01.007
[15]
Milnes, E.L., Thornton, G., Léveillé, A.N., Delnatte, P., Barta, J.R., Smith, D.A., et al. (2019) Babesia odocoilei and Zoonotic Pathogens Identified from Ixodes scapularis Ticks in Southern Ontario, Canada. TicksandTick-BorneDiseases, 10, 670-676. https://doi.org/10.1016/j.ttbdis.2019.02.016
Steiner, F.E., Pinger, R.R., Vann, C.N., Grindle, N., Civitello, D., Clay, K., et al. (2008) Infection and Co-Infection Rates of Anaplasma phagocytophilum Variants, Babesia Spp., Borrelia burgdorferi, and the Rickettsial Endosymbiont in Ixodes scapularis (Acari: Ixodidae) from Sites in Indiana, Maine, Pennsylvania, and Wisconsin. JournalofMedicalEntomology, 45, 289-297. https://doi.org/10.1093/jmedent/45.2.289
[18]
Waldrup, K.A., Kocan, A.A., Barker, R.W. and Wagner, G.G. (1990) Transmission of Babesia odocoilei in White-Tailed Deer (Odocoileus virginianus) by Ixodes scapularis (Acari: Ixodidae). JournalofWildlifeDiseases, 26, 390-391. https://doi.org/10.7589/0090-3558-26.3.390
[19]
Eshoo, M.W., Carolan, H.E., Massire, C., Chou, D.M., Crowder, C.D., Rounds, M.A., et al. (2015) Survey of Ixodes pacificus Ticks in California Reveals a Diversity of Microorganisms and a Novel and Widespread Anaplasmataceae Species. PLOSONE, 10, e0135828. https://doi.org/10.1371/journal.pone.0135828
[20]
Scott, J.D. and Scott, C.M. (2025) Molecular Detection of Borrelia burgdorferi Sensu Lato, Babesia odocoilei, Babesia microti and Anaplasma phagocytophilum in Ixodes Ticks in British Columbia, Canada. JournalofBiomedicalResearch&EnvironmentalSciences, 6, 1367-1379. https://doi.org/10.37871/jbres2192
[21]
Emerson, H.R. and Wright, W.T. (1968) The Isolation of a Babesia in White-Tailed Deer. BulletinoftheWildlifeDiseaseAssociation, 4, 142-143. https://doi.org/10.7589/0090-3558-4.4.142
[22]
Emerson, H.R. and Wright, W.T. (1970) Correction. Journal of Wildlife Diseases, 6, 519.
[23]
Waldrup, K.A., Kocan, A.A., Qureshi, T., Davis, D.S., Baggett, D. and Wagner, G.G. (1989) Serological Prevalence and Isolation of Babesia odocoilei among White-Tailed Deer (Odocoileus virginianus) in Texas and Oklahoma. JournalofWildlifeDiseases, 25, 194-201. https://doi.org/10.7589/0090-3558-25.2.194
[24]
Goff, W.L., Jessup, D.A., Waldrup, K.A., Thomford, J.W., Conrad, P.A., Boyce, W.M., et al. (1993) The Isolation and Partial Characterization of A Babesia Sp. from Desert Bighorn Sheep (Ovis canadensis Nelsoni). JournalofEukaryoticMicrobiology, 40, 237-243. https://doi.org/10.1111/j.1550-7408.1993.tb04909.x
[25]
Thomford, J.W., Conrad, P.A., Boyce, W.M., Holman, P.J. and Jessup, D.A. (1993) Isolation and in Vitro Cultivation of Babesia Parasites from Free-Ranging Desert Bighorn Sheep (Ovis Canadensis Nelsoni) and Mule Deer (Odocoileus hemionus) in California. TheJournalofParasitology, 79, 77-84. https://doi.org/10.2307/3283281
[26]
Scott, J.D., Fernando, K., Banerjee, S.N., Durden, L.A., Byrne, S.K., Banerjee, M., et al. (2001) Birds Disperse Ixodid (Acari: Ixodidae) and Borrelia burgdorferi-Infected Ticks in Canada. JournalofMedicalEntomology, 38, 493-500. https://doi.org/10.1603/0022-2585-38.4.493
[27]
Scott, J.D., Durden, L.A. and Anderson, J.F. (2015) Infection Prevalence of Borrelia burgdorferi in Ticks Collected from Songbirds in Far-Western Canada. OpenJournalofAnimalSciences, 5, 232-241. https://doi.org/10.4236/ojas.2015.53027
[28]
Scott, J.D., Clark, K. and Durden, L. (2019) Presence of Babesia odocoilei and Borrelia burgdorferi Sensu Stricto in a Tick and Dual Parasitism of Amblyomma inornatum and Ixodes scapularis on a Bird in Canada. Healthcare, 7, Article 46. https://doi.org/10.3390/healthcare7010046
[29]
Scott, J.D., Pascoe, E.L., Sajid, M.S. and Foley, J.E. (2020) Monitoring of Nesting Songbirds Detects Established Population of Blacklegged Ticks and Associated Lyme Disease Endemic Area in Canada. Healthcare, 8, Article 59. https://doi.org/10.3390/healthcare8010059
[30]
Scott, J.D., Pascoe, E.L., Sajid, M.S. and Foley, J.E. (2021) Detection of Babesia odocoilei in Ixodes scapularis Ticks Collected in Southern Ontario, Canada. Pathogens, 10, 327. https://doi.org/10.3390/pathogens10030327
[31]
Scott, J.D. and Pesapane, R.R. (2021) Detection of Anaplasma phagocytophilum, Babesia odocoilei, Babesia sp., Borrelia burgdorferi Sensu Lato, and Hepatozooncanis in Ixodes scapularis Ticks Collected in Eastern Canada. Pathogens, 10, Article 1265. https://doi.org/10.3390/pathogens10101265
[32]
Scott, J.D., McGoey, E. and Pesapane, R.R. (2022) Tick-borne Pathogens Anaplasma phagocytophilum, Babesia odocoilei, and Borrelia burgdorferi Sensu Lato in Blacklegged Ticks Widespread across Eastern Canada. JournalofBiomedicalResearch&EnvironmentalSciences, 3, 1249-1256. https://doi.org/10.37871/jbres1586
[33]
Scott, J.D., McGoey, E., Morales, A. and Pesapane, R.R. (2022) Molecular Detection of Anaplasma phagocytophilum, Babesia odocoilei, Babesia Species and Borrelia burgdorferi Sensu Lato in Songbirds. JournalofBiomedicalResearch&EnvironmentalSciences, 3, 1451-1459. https://doi.org/10.37871/jbres1619
[34]
Scott, J.D., Pascoe, E.L., Sajid, M.S. and Foley, J.E. (2020) Detection of Babesia odocoilei in Ixodes scapularis Ticks Collected from Songbirds in Ontario and Quebec, Canada. Pathogens, 9, Article 781. https://doi.org/10.3390/pathogens9100781
[35]
Scott, J.D., Foley, J.E., Anderson, J.F., Clark, K.L. and Durden, L.A. (2017) Detection of Lyme Disease Bacterium, Borrelia burgdorferi Sensu Lato, in Blacklegged Ticks Collected in the Grand River Valley, Ontario, Canada. InternationalJournalofMedicalSciences, 14, 150-158. https://doi.org/10.7150/ijms.17763
[36]
Keirans, J.E., Hutcheson, H.J., Durden, L.A. and Klompen, J.S.H. (1996) Ixodes (Ixodes) Scapularis (Acari: Ixodidae): Redescription of All Active Stages, Distribution, Hosts, Geographical Variation, and Medical and Veterinary Importance. JournalofMedicalEntomology, 33, 297-318. https://doi.org/10.1093/jmedent/33.3.297
[37]
Durden, L.A. and Keirans, J.E. (1996) Nymphs of the Genus Ixodes (Acari: Ixodidae) of the United States: Taxonomy, Identification Key, Distribution, Hosts, and Medical/Veterinary Importance. Entomological Society of America, 95 p.
[38]
Courtney, J.W., Kostelnik, L.M., Zeidner, N.S. and Massung, R.F. (2004) Multiplex Real-Time PCR for Detection of Anaplasma phagocytophilum and Borrelia burgdorferi. JournalofClinicalMicrobiology, 42, 3164-3168. https://doi.org/10.1128/jcm.42.7.3164-3168.2004
[39]
Tokarz, R., Tagliafierro, T., Cucura, D.M., Rochlin, I., Sameroff, S. and Lipkin, W.I. (2017) Detection of Anaplasma phagocytophilum, Babesia microti, Borrelia burgdorferi, Borrelia miyamotoi, and Powassan Virus in Ticks by a Multiplex Real-Time Reverse Transcription-PCR Assay. mSphere, 2, e00151-17. https://doi.org/10.1128/msphere.00151-17
[40]
Crandall, K.E., Kerr, J.T. and Millien, V. (2022) Emerging Tick-Borne Pathogens in Central Canada: Recent Detections of Babesia odocoilei and Rickettsiarickettsii. Vector-BorneandZoonoticDiseases, 22, 535-544. https://doi.org/10.1089/vbz.2022.0036
[41]
Holden, K., Boothby, J.T., Anand, S. and Massung, R.F. (2003) Detection of Borrelia burgdorferi, Ehrlichiachaffeensis, and Anaplasma phagocytophilum in Ticks (Acari: Ixodidae) from a Coastal Region of California. JournalofMedicalEntomology, 40, 534-539. https://doi.org/10.1603/0022-2585-40.4.534
[42]
Persing, D.H., Mathiesen, D., Marshall, W.F., Telford, S.R., Spielman, A., Thomford, J.W., et al. (1992) Detection of Babesia microti by Polymerase Chain Reaction. JournalofClinicalMicrobiology, 30, 2097-2103. https://doi.org/10.1128/jcm.30.8.2097-2103.1992
[43]
Burgess, H.J., Lockerbie, B.P., Ayalew, L.E., Dibernardo, A., Hrazdilová, K., Modry, D., et al. (2021) Species-Specific PCR Assay for the Detection of Babesia odocoilei. JournalofVeterinaryDiagnosticInvestigation, 33, 1188-1192. https://doi.org/10.1177/10406387211032927
[44]
Rajakaruna, R.S., Capps-Ludwig, D., Durden, L.A. and Eremeeva, M.E. (2025) Detection of Rickettsia and Bartonella in Fleas and Ticks Collected from Pets at Veterinary Clinics in Georgia, United States. JournalofParasitology, 111, 113-122. https://doi.org/10.1645/24-109
[45]
Newbold, C., Craig, A., Kyes, S., Rowe, A., Fernandez-Reyes, D. and Fagan, T. (1999) Cytoadherence, Pathogenesis and the Infected Red Cell Surface in Plasmodiumfalciparum. InternationalJournalforParasitology, 29, 927-937. https://doi.org/10.1016/s0020-7519(99)00049-1
[46]
Schetters, T. (2019) Mechanisms Involved in the Persistence of Babesia Canis Infection in Dogs. Pathogens, 8, Article 94. https://doi.org/10.3390/pathogens8030094
[47]
Scott, J.D. and Scott, C.M. (2025) Zoonotic Pathogens Anaplasma phagocytophilum, Babesia microti, Babesia odocoilei, and Borrelia burgdorferi Sensu Lato Detected in Ixodes scapularis Ticks Collected at an Established Population in Eastern Canada. JournalofBiomedicalResearch&EnvironmentalSciences, 6, 1852-1864. https://doi.org/10.37871/jbres2233
[48]
Roberts, E.D., Bohm Jr., R.P., Cogswell, R.B., Lanners, H.N., Lowrie, R.E. and Povinelli, L. (1995) Chronic Lyme Disease in the Rhesus Monkey. LaboratoryInvestigation, 72, 146-160.
[49]
Stricker, R.B., Fesler, M.C. and Johnson, L. (2026) Persistent Borrelia Infection in Chronic Lyme Disease: A Review of the Medical Literature. AdvancesinInfectiousDiseases, 16, 73-87. https://doi.org/10.4236/aid.2026.161005
[50]
Scott, J.D., Clark, K.L., Foley, J.E., Bierman, B.C. and Durden, L.A. (2018) Far-Reaching Dispersal of Borrelia burgdorferi Sensu Lato-Infected Blacklegged Ticks by Migratory Songbirds in Canada. Healthcare, 6, Article 89. https://doi.org/10.3390/healthcare6030089
[51]
Scott, J.D., Clark, K.L., Foley, J.E., Anderson, J.F., Bierman, B.C. and Durden, L.A. (2018) Extensive Distribution of the Lyme Disease Bacterium, Borrelia burgdorferi Sensu Lato, in Multiple Tick Species Parasitizing Avian and Mammalian Hosts across Canada. Healthcare, 6, Article 131. https://doi.org/10.3390/healthcare6040131
[52]
Scott, J.D. and Scott, C.M. (2023) Primary Detection of the Establishment of Blacklegged Ticks, Ixodes scapularis, in British Columbia, Canada. JournalofBiomedicalResearch&EnvironmentalSciences, 4, 935-941. https://doi.org/10.37871/jbres1754
[53]
Scott, J.D., McKeown, J.T.A. and Scott, C.M. (2023) Migratory Songbirds Transport Amblyommalongirostre and Amblyommamaculatum Ticks to Canada. JournalofBiomedicalResearch&EnvironmentalSciences, 4, 150-156. https://doi.org/10.37871/jbres1659
[54]
Scott, J.D., Anderson, J.F. and Durden, L.A. (2012) Widespread Dispersal of Borrelia burgdorferi-Infected Ticks Collected from Songbirds across Canada. JournalofParasitology, 98, 49-59. https://doi.org/10.1645/ge-2874.1
[55]
Richter, D., Spielman, A., Komar, N. and Matuschka, F. (2000) Competence of American Robins as Reservoir Hosts for Lyme Disease Spirochetes. EmergingInfectiousDiseases, 6, 133-138. https://doi.org/10.3201/eid0602.000205
