Persian Journal of Acarology

Persian Journal of Acarology

Molecular detection of Hepatozoon canis(Apicomplexa: Hepatozoidae)in ticks (Ixodida) collected from dogs in Kerman, Iran

Document Type : Original Article

Authors
1 Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Bahonar University, Kerman, Iran
2 Department of Pathobiology, Faculty of Veterinary Medicine, Islamic Azad University, Science and Research Branch, Tehran, Iran
3 Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Bahonar University, Kerman, Iran
4 Department of Pathobiology, Faculty of Veterinary Science, Bu-Ali Sina University, Hamedan, Iran
Abstract
Previous studies from Iran showed that canine hepatozoonosis is prevalent in dogs. Still, there is limited information about the tick vectors of Hepatozoon canis (Apicomplexa: Hepatozoidae) although Ripicephalus sanguineus sensu lato complex ticks (Acari: Ixodida) are known to be competent vectors. In Iran, He. canis was detected in R. sanguineus and R. turanicus in the west of the country. This study aimed to investigate the molecular prevalence of He. canis in ticks collected from stray dogs in a southeastern city of Iran. From 2019 to 2020, 50 tick-infested free-ranging dogs (24 female, 26 male) in Kerman city were randomly chosen. From each dog, five ticks were collected in individually labeled tubes and transferred to the laboratory. Ticks of each dog were identified morphologically, crushed, and pooled for genomic DNA extraction. Conventional PCR targeting an 891 bp fragment of the 18S ribosomal RNA gene of He. canis was performed on 50 tick pools, and products were gel-electrophoresed. All of the collected ticks were diagnosed as R. sanguineus sensu lato. DNA of He. canis was detected in 14 tick pools (28%). Dogs from which positive ticks were collected were of both sexes and aged between 5 months and 6 years old. This study shows for the first time a considerable prevalence of He. canis in ticks feeding from stray dogs of Kerman implying a potential health risk for domestic and wild canine hosts in the region. It also shows that molecular examination of ticks for Hepatozoon DNA could be a proper alternative to blood collection in epidemiological studies. Effective ectoparasite control strategies, regular examination of dogs, and successful chemoprophylaxis are advocated.
Keywords

Alanazi, A.D., Nguyen, V.L., Alyousif, M.S., Manoj, R.R.S., Alouffi, A.S., Donato, R., Sazmand, A., Mendoza-Roldan, J.A., Dantas-Torres, F. & Otranto, D. (2020) Ticks and associated pathogens in camels (Camelus dromedarius) from Riyadh Province, Saudi Arabia. Parasites & Vectors, 13(1): 110. DOI: 10.1186/s13071-020-3973-y
Andersson, M.O., Tolf, C., Tamba, P., Stefanache, M., Radbea, G., Rubel, F., Waldenström, J., Dobler, G. & Chițimia-Dobler, L. (2017) Babesia, Theileria, and Hepatozoon species in ticks infesting animal hosts in Romania. Parasitology Research, 116: 2291–2297.
Bahiraei, Z., Sazmand, A., Nourian, A. & Otranto, D. (2023) Identification of Hepatozoon and Cercopithifilaria in ticks infesting dogs of different regions of Iran. Journal of Entomological Society of Iran, 43(1): 53–61. DOI: 10.52547/jesi.43.1.6
Baneth, G. & Allen, K. (2022) Hepatozoonosis of dogs and cats. Veterinary Clinics: Small Animal Practice, 52(6): 1341–1358.
Baneth, G., Samish, M., Aleksefv, E., Aroch, I. & Shkap, V. (2001) Transmission of Hepatozoon canis to dogs by naturally-fed or percutaneously-injected Rhipicephalus sanguineus ticks. Journal of Parasitology, 87(3): 606–611. DOI: 10.1645/0022-3395(2001)087[0606:TOHCTD] 2.0.CO;2
Barati, A. & Razmi, G.R. (2018) A parasitologic and molecular survey of Hepatozoon canis infection in stray dogs in Northeastern Iran. Journal of Parasitology, 104(4): 413–417. DOI: 10.1645/17-105
Beikpour, F., Ndiana, L.A., Sazmand, A., Capozza, P., Nemati, F., Pellegrini, F., Zafari, S., Zolhavarieh, SM., Cardone, R. & Faraji, R. (2022) Detection and genomic characterization of canine circovirus in Iran. Animals, 12(4): 507. DOI: 10.3390/ani12040507
Beugnet, F., Halos, L. & Guillot, J. (2018) Textbook of clinical parasitology in dogs and cats. Grupo Asís Biomedia, S.L., Zaragoza, Spain, 413 pp.
Beus, K., Goudarztalejerdi, A. & Sazmand A. (2024) Molecular detection and identification of hemotropic Mycoplasma species in dogs and their ectoparasites in Iran. Scientific Reports, 14(1): 580. DOI: 10.1038/s41598-024-51173-w
Dalimi, A., Jameie, F., Mohammadiha, A., Barati, M. & Molaei, S. (2017) Molecular detection of Hepatozoon canis in dogs of Ardabil Province, Northwest of Iran. Archives of Razi Institute, 72(3): 197–201. DOI: 10.22034/ari.2017.108389
Dantas-Torres, F. (2010) Biology and ecology of the brown dog tick, Rhipicephalus sanguineus. Parasites & Vectors, 3(1): 26. DOI: 10.1186/1756-3305-3-26
Dantas-Torres, F., Chomel, B.B. & Otranto D. (2012) Ticks and tick-borne diseases: a One Health perspective. Trends in Parasitology, 28(10): 437–446. DOI: 10.1016/j.pt.2012.07.003
Dantas-Torres, F. & Otranto, D. (2022) Rhipicephalus sanguineus (Brown dog tick). Trends in Parasitology, 38(11): 993–994. DOI: 10.1016/j.pt.2022.08.011
Del Cerro, A., Oleaga, A., Somoano, A., Barandika, J.F., García-Pérez, A.L. & Espí, A. (2022) Molecular identification of tick-borne pathogens (Rickettsia spp., Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Coxiella burnetii and piroplasms) in questing and feeding hard ticks from North-Western Spain. Ticks and Tick-Borne Diseases, 13(4): 101961. DOI: 10.1016/j. ttbdis.2022.101961
Estrada-Peña, A., Mihalca, A.D. & Petney, T.N. (Eds.) (2018) Ticks of Europe and North Africa: a guide to species identification. Springer, Cham., 404 pp.
Farkas, R., Solymosi, N., Takács, N., Hornyák, Á., Hornok, S., Nachum-Biala, Y. & Baneth, G. (2014) First molecular evidence of Hepatozoon canis infection in red foxes and golden jackals from Hungary. Parasites & Vectors, 7: 1–7. DOI: 10.1186/1756-3305-7-303
Geurden, T., Becskei, C., Six, R.H., Maeder, S., Latrofa, M.S., Otranto, D. & Farkas, R. (2018) Detection of tick-borne pathogens in ticks from dogs and cats in different European countries. Ticks and Tick-Borne Diseases, 9(6): 1431–1436. DOI: 10.1016/j.ttbdis.2018.06.013
Ghodrati, S., Lesiczka, P.M., Zurek, L., Szekely, F. & Modrý, D. (2024) Rhipicephalus sanguineus from Hungarian dogs: Tick identification and detection of tick-borne pathogens. Veterinary Parasitology: Regional Studies and Reports, 50: 101007. DOI: 10.1016/j.vprsr.2024.101007
Giannelli, A., Lia, R.P., Annoscia, G., Buonavoglia, C., Lorusso, E., Dantas-Torres, F., Baneth, G. & Otranto, D. (2017) Rhipicephalus turanicus, a new vector of Hepatozoon canis. Parasitology, 144(6): 730–737. DOI: 10.1017/S003118201600250X
Greay, T.L., Zahedi, A., Krige, A.S., Owens, J.M., Rees, R.L., Ryan, U.M., Oskam, C.L. & Irwin, P.J. (2018) Endemic, exotic and novel apicomplexan parasites detected during a national study of ticks from companion animals in Australia. Parasites & Vectors, 11: 1–20. DOI: 10.1186/s130 71-018-2775-y
Guo, W.P., Xie, G.C., Xue, Z.Q., Yu, J.J., Jian, R., Du, L.Y. & Li, Y.N. (2020) Molecular detection of Hepatozoon canis in dogs and ticks in Shaanxi province, China. Comparative Immunology, Microbiology and Infectious Diseases, 72: 101514. DOI: 10.1016/j.cimid.2020.101514
Hamšíková, Z., Silaghi, C., Rudolf, I., Venclíková, K., Mahríková, L., Slovák, M., Mendel, J., Blažejová, H., Berthová, L. & Kocianová, E. (2016) Molecular detection and phylogenetic analysis of Hepatozoon spp. in questing Ixodes ricinus ticks and rodents from Slovakia and Czech Republic. Parasitology Research, 115: 3897–3904. DOI: 10.1007/s00436-016-5156-5
Hornok, S., Tánczos, B., de Mera, I.G.F., de la Fuente, J., Hofmann-Lehmann, R. & Farkas, R. (2013) High prevalence of Hepatozoon-infection among shepherd dogs in a region considered to be free of Rhipicephalus sanguineus. Veterinary Parasitology, 196(1–2): 189–193. DOI: 10.1016/j.vetp ar.2013.02.009
Hosseini, S.H., Manshori-Ghaishghorshagh, F., Ramezani, M., Nayebzadeh, H., Ahoo, M.B., Eslamian, A., Soltani, M., Jamshidi, S., Bezerra-Santos, M.A. & Jalousian, F. (2022) Canine microfilaraemia in some regions of Iran. Parasites & Vectors, 15(1): 90. DOI: 10.1186/s13071-022-05209-7
Hosseini-Chegeni, A., Nasrabadi, M., Sadat Hashemi-Aghdam, S., Oshaghi, MA., Lotfi, A., Telmadarraiy, Z. & Sedaghat, MM. (2019a) Molecular identification of Rhipicephalus species (Acari: Ixodidae) parasitizing livestock from Iran. Mitochondrial DNA Part A, 30(3): 448–456. DOI: 10.1080/24701394.2018.1546298
Hosseini-Chegeni, A., Tavakoli, M. & Telmadarraiy, Z. (2019b) The updated list of ticks (Acari: Ixodidae & Argasidae) occurring in Iran with a key to the identification of species. Systematic and Applied Acarology, 24(11): 2133–2166. DOI: 10.11158/saa.24.11.8
Iatta, R., Sazmand, A., Nguyen, V.L., Nemati, F., Ayaz, M.M., Bahiraei, Z., Zafari, S., Giannico, A., Greco, G. & Dantas-Torres, F. (2021) Vector-borne pathogens in dogs of different regions of Iran and Pakistan. Parasitology Research, 120(12): 4219–4228 DOI: 10.1007/s00436-020-0699 2-x
Jamshidi, S., Maazi, N., Ranjbar-Bahadori, S., Rezaei, M., Morakabsaz, P. & Hosseininejad, M. (2012) A survey of ectoparasite infestation in dogs in Tehran, Iran. Revista Brasileira de Parasitologia Veterinária, 21(3): 326–329. DOI: 10.1590/S1984-29612012000300030
Jarquín-Díaz, V.H., Barbachano-Guerrero, A., Maldonado-Rodríguez, R., Vásquez-Aguilar, A.A. & Aguilar-Faisal, J.L. (2016) First molecular evidence of Hepatozoon canis in domestic dogs and ticks in fragmented rainforest areas in Mexico. Veterinary Parasitology: Regional Studies and Reports, 6: 4–8. DOI: 10.1016/j.vprsr.2016.11.001
Kamani, J., Baneth, G., Mumcuoglu, K.Y., Waziri, N.E., Eyal, O., Guthmann, Y. & Harrus, S. (2013) Molecular detection and characterization of tick-borne pathogens in dogs and ticks from Nigeria. PLoS Neglected Tropical Diseases, 7(3): e2108. DOI: 10.1371/journal.pntd.0002108
Karasartova, D., Gureser, A.S., Gokce, T., Celebi, B., Yapar, D., Keskin, A., Celik, S., Ece, Y., Erenler, A.K. & Usluca, S. (2018) Bacterial and protozoal pathogens found in ticks collected from humans in Corum province of Turkey. PLoS Neglected Tropical Diseases, 12(4): e000639 5. DOI: 10.1371/journal.pntd.0006395
Khazeni, A., Telmadarraiy, Z., Oshaghi, M.A., Mohebali, M., Zarei, Z. & Abtahi, SM. (2013) Molecular detection of Ehrlichia canis in ticks population collected on dogs in Meshkin-Shahr, Ardebil Province, Iran. Journal of Biomedical Science and Engineering, 6: 1–5. DOI: 10.4236/jbi se.2013.67A3001
Khoshnegah, J., Mohri, M., Movassaghi, A.R. & Kazemi Mehrjerdi, H. (2009) The first report of Hepatozoon canis infection of a dog in Iran. Comparative Clinical Pathology, 18(4): 455–458. DOI: 10.1007/s00580-008-0794-3
Laidoudi, Y., Bedjaoui, S., Medkour, H., Latrofa, M.S., Mekroud, A., Bitam, I., Davoust, B., Otranto, D. & Mediannikov, O. (2020) Molecular approach for the diagnosis of blood and skin canine filarioids. Microorganisms, 8(11): 1671. DOI: 10.3390/microorganisms8111671
Mirani, F., Yakhchali, M. & Naem, S. (2017) A study on ectoparasites fauna of dogs in suburbs of Ghilanegharb, Kermanshah province, Iran. Journal of Faculty of Veterinary Medicine University of Tehran, 72(1): 7–14.
Mitková, B., Hrazdilová, K., Steinbauer, V., D’Amico, G., Mihalca, A.D. & Modrý, D. (2016) Autochthonous Hepatozoon infection in hunting dogs and foxes from the Czech Republic. Parasitology Research, 115: 4167–4171. DOI: 10.1007/s00436-016-5191-2
Mostafavi, S.M., Khalili, M., Akhtardanesh, B., Nourollahifard, S.R. & Esmaeili, S. (2022) Rickettsia spp. in Rhipicephalus sanguineus sensu lato ticks collected from stray dogs in Kerman city, Iran. Ticks and Tick-Borne Diseases, 13(5): 101985. DOI: 10.1016/j.ttbdis.2022.101985
Murata, T., Inoue, M., Tateyama, S., Taura, Y. & Nakama, S. (1993) Vertical transmission of Hepatozoon canis in dogs. Journal of Veterinary Medical Science, 55(5): 867–868. DOI: 10.1292 /jvms.55.867
Myers, S., Clow, K., DeWinter, S., Sundstrom, K. & Little, S. (2024) Multiple species of canine Rhipicephalus complex detected in Canada. Veterinary Parasitology: Regional Studies and Reports, 48: 100976. DOI: 10.1016/j.vprsr.2023.100976
Najm, N.A., Meyer-Kayser, E., Hoffmann, L., Pfister, K. & Silaghi, C. (2014) Hepatozoon canis in German red foxes (Vulpes vulpes) and their ticks: molecular characterization and the phylogenetic relationship to other Hepatozoon spp. Parasitology Research, 113: 2679–2685. DOI: 10.1007/s00436-014-3923-8
Nava, S., Beati, L., Venzal, J.M., Labruna, M.B., Szabó, M.P., Petney, T., Saracho-Bottero, M.N., Tarragona, E.L., Dantas-Torres, F. & Silva, M.M.S. (2018) Rhipicephalus sanguineus (Latreille, 1806): Neotype designation, morphological re-description of all parasitic stages and molecular characterization. Ticks and Tick-Borne Diseases, 9(6): 1573–1585. DOI: 10.1016/j.ttbdis.2018.0 8.001
Orkun, Ö. & Emir, H. (2020) Identification of tick-borne pathogens in ticks collected from wild animals in Turkey. Parasitology Research, 119(9): 3083–3091. DOI: 10.1007/s00436-020-0681 2-2
Otranto, D., Huchet, J.B., Giannelli, A., Callou, C. & Dantas-Torres, F. (2014) The enigma of the dog mummy from Ancient Egypt and the origin of ‘Rhipicephalus sanguineus’. Parasites & Vectors, 2: 7. DOI: 10.1186/1756-3305-7-2
Rahmani Amoli, A., Khoshnegah, J. & Razmi, G.R. (2012) A preliminary parasitological survey of Hepatozoon spp. infection in dogs in Mashhad, Iran. Iranian Journal of Parasitology, 7(4): 99–103.
Sazmand, A., Bahiraei, Z., Nemati, F., Annoscia, G., Bezerra-Santos, M.A., Nayebzadeh, H., Salemi, AM., Mousavi, SM., Sadjjadi, SM. & Otranto, D. (2022) Dermal microfilariae of dogs, jackals and cats in different regions of Iran. Parasites & Vectors, 15(1): 28. DOI: 10.1186/s13071-021-05141-2
Sazmand, A., Eigner, B., Mirzaei, M., Hekmatimoghaddam, S.H., Harl, J., Duscher, G.G., Fuehrer, H.P. & Joachim, A. (2016) Molecular identification of hemoprotozoan parasites in camels (Camelus dromedarius) of Iran. Iranian Journal of Parasitology, 11(4): 568–573.
Schäfer, I., Müller, E., Nijhof, A.M., Aupperle-Lellbach, H., Loesenbeck, G., Cramer, S. & Naucke, T.J. (2022) First evidence of vertical Hepatozoon canis transmission in dogs in Europe. Parasites & Vectors, 15(1): 296. DOI: 10.1186/s13071-022-05392-7
Shamshiri, Z., Goudarztalejerdi, A., Zolhavarieh, S.M., Kamalpour, M. & Sazmand, A. (2023) Molecular identification of Bartonella species in dogs and arthropod vectors in Hamedan and Kermanshah, Iran. Iranian Veterinary Journal, 19(2): 104–116. DOI: 10.22055/ivj.2022.325115 .2436
Shiri, A., Kheirandish, F., Sazmand, A., Kayedi, M.H. & Hosseini-Chegeni, A. (2024) Molecular identification of hemoparasites in ixodid ticks in Iran. Veterinary Parasitology: Regional Studies and Reports, 47: 100967. DOI: 10.1016/j.vprsr.2023.100967
Šlapeta, J., Chandra, S. & Halliday, B. (2021) The “tropical lineage” of the brown dog tick Rhipicephalus sanguineus sensu lato identified as Rhipicephalus linnaei. International Journal for Parasitology, 51(6): 431–436. DOI: 10.1016/j.ijpara.2021.02.001
Šlapeta, J., Halliday, B., Chandra, S., Alanazi, A.D. & Abdel-Shafy, S. (2022) Rhipicephalus linnaei (Audouin, 1826) recognised as the “tropical lineage” of the brown dog tick Rhipicephalus sanguineus sensu lato: neotype designation, redescription, and establishment of morphological and molecular reference. Ticks and Tick-Borne Diseases, 13(6): 102024. DOI: 10.1016/j.ttbdis. 2022.102024
Šlapeta, J., Halliday, B., Dunlop, J.A., Nachum-Biala, Y., Salant, H., Ghodrati, S., Modrý, D. & Harrus, S. (2023) The “southeastern Europe” lineage of the brown dog tick Rhipicephalus sanguineus (sensu lato) identified as Rhipicephalus rutilus Koch, 1844: Comparison with holotype and generation of mitogenome reference from Israel. Current Research in Parasitology & Vector Borne Diseases, 3: 100118. DOI: 10.1016/j.crpvbd.2023.100118
Smith, T.G. (1996) The genus Hepatozoon (Apicomplexa: Adeleina). Journal of Parasitology, 1: 565–585.
Soltani, R. & Dalimi, A. (2018) A molecular study on Hepatozoon canis infection in dogs in Tehran (Iran). Archives of Razi Institute, 73(4): 257–263. DOI: 10.22092/ari.2017.110293.1125
Tolnai, Z., Sréter-Lancz, Z. & Sréter, T. (2015) Spatial distribution of Anaplasma phagocytophilum and Hepatozoon canis in red foxes (Vulpes vulpes) in Hungary. Ticks and Tick-Borne Diseases, 6(5): 645–648. DOI: 10.1016/j.ttbdis.2015.05.009
Zeinali, M., Malekifard, F., Rakhshanpour, A. & Yakhchali, M. (2022) Study on Hepatozoon canis in dogs in Urmia region. Iranian Journal of Animal Science, 52(4): 253–260. DOI: 10.22059/ijas. 2021.329716.653845