Molecular characterization of Hepatozoon spp. in naturally infected dogs in Aydın province

Metin Pekağırbaş; Muhammed Veli Demirbilek; Emrah Simsek; Heycan Berk Aydın; Hakan Kanlıoğlu; Asude Gülçe Oryaşın; Nuran Selek

doi:10.33988/auvfd.1561161

Research Article

Molecular characterization of Hepatozoon spp. in naturally infected dogs in Aydın province

Year 2025, Accepted Papers, 1 - 6

Metin Pekağırbaş , Muhammed Veli Demirbilek , Emrah Simsek , Heycan Berk Aydın , Hakan Kanlıoğlu , Asude Gülçe Oryaşın , Nuran Selek

https://doi.org/10.33988/auvfd.1561161

Abstract

Canine hepatozoonosis, a disease caused by the protozoan Hepatozoon canis and Hepatozoon americanum, represents a significant tick-borne disease affecting domestic and wild carnivores. The objective of this study was to detect Hepatozoon species in randomly selected dogs from Aydın by PCR and to elucidate their molecular characterization and phylogenetic differences through sequence analysis. In total, 100 blood samples collected from dogs were analyzed, and the prevalence of Hepatozoon DNA was determined to be 3%, with only three samples testing positive. Partial sequences of the 18S rRNA gene exhibited 100% similarity with corresponding H. canis isolates. Phylogenetic analysis of the 18S rRNA gene region revealed the formation of two primary clusters, one consisting of H. canis isolates and the other comprising different Hepatozoon species. While H. canis isolates formed distinct subclusters, they were all grouped separately from other Hepatozoon species. Furthermore, phylogenetic analysis highlighted the presence of multiple H. canis haplotypes in Türkiye, with intraspecific nucleotide differences ranging from 0.0% to 2.9%. The nucleotide differences among the isolates identified in this study ranged from 0.0% to 1.6%. All sequences obtained in this study have been submitted to GenBank and assigned accession numbers PQ669652, PQ671331 and PQ669658. These findings underscore the need for further investigations into Hepatozoon infections among cats and wild animals in the region. Additionally, the detection of the parasite in vector ticks could offer valuable insights into the genetic diversity and distribution of circulating Hepatozoon species.

Keywords

Aydın, Dogs, Hepatozoon canis, Molecular Characterization

Ethical Statement

Ethical approval for this study was obtained from the Aydın Adnan Menderes University Local Ethics Committee (Decision number 64583101/2018/069).

Supporting Institution

This research received no grant from any funding agency/sector.

References

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4. Aslan B, Çelik ÖY, Ayan A, et al (2022): A Molecular survey of Hepatozoon canis in dogs in the Siirt province of Turkey. Acta Veterinaria Brno, 91, 277-283.
5. Aydin MF, Sevinc F, Sevinc M (2015): Molecular detection and characterization of Hepatozoon spp. in dogs from the Central part of Turkey. Ticks Tick borne Dis, 6, 388-392.
6. Aysul N, Ural K, Ulutas B et al (2013): First detection and molecular identification of Babesia gibsoni in two dogs from the Aydin Province of Turkey. Turk J of Vet Anim Sci, 37, 226-229
7. Bakırcı S, Aysul N, Bilgiç HB, et al (2019): Tick Bites on Humans in Southwestern Region of Turkey: Species Diversity. Türkiye Parasitol Derg, 43, 30.
8. Baneth G, Vincent-Johnson N (2005): Hepatozoonosis. In: Shaw, S.E., Day, M.J. (Eds.), Arthropod-borne Infectious Diseases of the Dog and Cat. Manson Publishing, London, pp. 78–88.
9. Bolukbas CS, Pekmezci D, Gurler AT, et al (2016): Molecular survey of Hepatozoon canis in dogs from Samsun Province of Northern part of Turkey. Etlik Vet Mikrobiyol Derg, 27, 104-107.
10. Bouattour A, Chabchoub A, Hajjaji I et al (2021): Hepatozoon canis and Babesia vogeli infections of dogs in Tunisia. Vet Parasitol Reg Stud Reports. 23, 100512.
11. Bowman DD (2009): Parasitology for Veterinarians. Saunders Elsevier, Ninth Edition, St. Louis, Missori.
12. Dumler JS, Barbet AF, Bekker CP, et al (2001): Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and “HGE agent” as subjective synonyms of Ehrlichia phagocytophila. Int J Syst Evol Microbiol, 51, 2145–2165.
13. Düzlü Ö, İnci A, Yıldırım A, et al (2014): The investigation of some tick-borne protozoon and rickettsial infections in dogs by Real Time PCR and the molecular characterizations of the detected isolates. Ankara Univ Vet Fak Derg, 61, 275-282.
14. Guven E, Avcioglu H, Cengiz S, et al (2017): Vector-borne pathogens in stray dogs in Northeastern Turkey. Vector Borne Zoonotic Dis 17, 610-617.
15. Hasegawa M, Iida Y, Yano T, et al (1985): Phylogenetic relationships among eukaryotic kingdoms inferred from ribosomal RNA sequences. J Mol Evol, 22, 32-38.
16. Inokuma H, Okuda M, Ohno K, et al (2002): Analysis of the 18S rRNA gene sequence of a Hepatozoon detected in two Japanese dogs. Vet Parasitol, 106, 265-271.
17. Karagenc TI, Pasa S, Kirli G, et al (2006): A parasitological, molecular and serological survey of Hepatozoon canis infection in dogs around the Aegean coast of Turkey. Vet Parasitol, 135, 113-119.
18. Kimura M (1980): A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol, 16, 111-20.
19. Kumar S, Stecher G, Li M, et al (2018): MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Mol Biol Evol, 35, 1547–1549.
20. Majlathova V, Hurnikova Z, Majlath I, et al (2007): Hepatozoon canis infection in Slovakia: imported or autochthonous? Vector Borne Zoonotic Dis, 7, 199-202.
21. Matsuu A, Ono S, Ikadai H, et al (2005): Development of a SYBR green real-time polymerase chain reaction assay for quantitative detection of Babesia gibsoni (Asian genotype) DNA. Journal Veterinary Diagn Invest, 17, 569-573. 22. Najm NA, Meyer-Kayser E, Hoffmann L, et al (2014): Hepatozoon canis in German red foxes (Vulpes vulpes) and their ticks: molecular characterization and the phylogenetic relationship to other Hepatozoon spp. Parasitol Res, 113, 2679-2685. 23. Ogden NH, Lindsay LR (2016): Effects of Climate and Climate Change on Vectors and Vector-Borne Diseases: Ticks Are Different. Trends Parasitol, 32, 646–656.
24. Orkun Ö, Koç N, Sürsal N, et al (2018): Molecular characterization of tick-borne blood protozoa in stray dogs from Central Anatolia Region of Turkey with a high-rate Hepatozoon infection. Kafkas Univ Vet Fak Derg, 24, 227-232, 2018.
25. Orkun Ö, Nalbantoğlu S (2018): Hepatozoon canis in Turkish red foxes and their ticks. Vet Parasitol Reg Stud Rep, 13, 35–37.
26. Otranto D, Dantas-Torres F, Breitschwerdt EB (2009): Managing canine vector-borne diseases of zoonotic concern: Part two. Trends Parasitol, 25, 228–235.
27. Otranto D, Dantas-Torres F, Weigl S, et al (2011): Diagnosis of Hepatozoon canis in young dogs by cytology and PCR. Parasit Vectors 4, 55.
28. Rochlin I, Toledo A (2020): Emerging tick-borne pathogens of public health importance: a mini-review. J Med Microbiol, 69, 781-791.
29. Stich RW, Blagburn BL, Bowman DD, et al (2014): Quantitative factors proposed to influence the prevalence of canine tick-borne disease agents in the United States. Parasit Vectors, 7, 417.
30. Sutherst RW (2004): Global change and human vulnerability to vectorborne diseases. Clin Microbiol Rev, 17, 136–173.
31. Tuna GE, Bakirci S, Ulutaş B (2020): Evaluation of clinical and haematological findings of mono-and co-infection with Hepatozoon canis in dogs. Animal Health Prod Hyg, 9, 696-702
32. Tuzdil AN (1933): Bizde ilk defa görülen bir Hepatozoon canis vakası. Türk Bay Cem Mec, 13, 35.
33. Ul-Hasan M, Abubakar M, Muhammad G, et al (2012): Prevalence of tick infestation (Rhipicephalus sanguineus and Hyalomma anatolicum anatolicum) in dogs in Punjab, Pakistan. Vet Italia, 48, 95–98
34. Vincent-Johnson NA, Macintire DK, Lindsay DL, et al (1997): A new Hepatozoon species from dogs: description of the causative agent of canine hepatozoonosis in North America. J Parasitol, 83, 1165–1172. 35. Vojta L, Mrljak V, Ćurković S, et al (2009): Molecular epizootiology of canine hepatozoonosis in Croatia. Int J Parasitol, 39, 1129–1136 36. Voyvoda H, Pasa S, Uner A (2004): Clinical Hepatozoon canis infection in a dog in Turkey. J Small Anim Pract, 45, 613-61.

Year 2025, Accepted Papers, 1 - 6

Metin Pekağırbaş , Muhammed Veli Demirbilek , Emrah Simsek , Heycan Berk Aydın , Hakan Kanlıoğlu , Asude Gülçe Oryaşın , Nuran Selek

https://doi.org/10.33988/auvfd.1561161

Abstract

References

1.Ahmad AS, Saeed MA, Rashid I, et al (2018): Molecular characterization of Hepatozoon canis from farm dogs in Pakistan. Parasitol Res, 117, 1131-1138.
2. Aktas M, Ozubek S, Altay K, et al (2015): A molecular and parasitological survey of Hepatozoon canis in domestic dogs in Turkey. Vet Parasitol, 209, 264-267.
3. Aktas M, Ozubek S, Ipek DNS (2013): Molecular investigations of Hepatozoon species in dogs and developmental stages of Rhipicephalus sanguineus. Parasitol Res, 112, 2381-2385.
4. Aslan B, Çelik ÖY, Ayan A, et al (2022): A Molecular survey of Hepatozoon canis in dogs in the Siirt province of Turkey. Acta Veterinaria Brno, 91, 277-283.
5. Aydin MF, Sevinc F, Sevinc M (2015): Molecular detection and characterization of Hepatozoon spp. in dogs from the Central part of Turkey. Ticks Tick borne Dis, 6, 388-392.
6. Aysul N, Ural K, Ulutas B et al (2013): First detection and molecular identification of Babesia gibsoni in two dogs from the Aydin Province of Turkey. Turk J of Vet Anim Sci, 37, 226-229
7. Bakırcı S, Aysul N, Bilgiç HB, et al (2019): Tick Bites on Humans in Southwestern Region of Turkey: Species Diversity. Türkiye Parasitol Derg, 43, 30.
8. Baneth G, Vincent-Johnson N (2005): Hepatozoonosis. In: Shaw, S.E., Day, M.J. (Eds.), Arthropod-borne Infectious Diseases of the Dog and Cat. Manson Publishing, London, pp. 78–88.
9. Bolukbas CS, Pekmezci D, Gurler AT, et al (2016): Molecular survey of Hepatozoon canis in dogs from Samsun Province of Northern part of Turkey. Etlik Vet Mikrobiyol Derg, 27, 104-107.
10. Bouattour A, Chabchoub A, Hajjaji I et al (2021): Hepatozoon canis and Babesia vogeli infections of dogs in Tunisia. Vet Parasitol Reg Stud Reports. 23, 100512.
11. Bowman DD (2009): Parasitology for Veterinarians. Saunders Elsevier, Ninth Edition, St. Louis, Missori.
12. Dumler JS, Barbet AF, Bekker CP, et al (2001): Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and “HGE agent” as subjective synonyms of Ehrlichia phagocytophila. Int J Syst Evol Microbiol, 51, 2145–2165.
13. Düzlü Ö, İnci A, Yıldırım A, et al (2014): The investigation of some tick-borne protozoon and rickettsial infections in dogs by Real Time PCR and the molecular characterizations of the detected isolates. Ankara Univ Vet Fak Derg, 61, 275-282.
14. Guven E, Avcioglu H, Cengiz S, et al (2017): Vector-borne pathogens in stray dogs in Northeastern Turkey. Vector Borne Zoonotic Dis 17, 610-617.
15. Hasegawa M, Iida Y, Yano T, et al (1985): Phylogenetic relationships among eukaryotic kingdoms inferred from ribosomal RNA sequences. J Mol Evol, 22, 32-38.
16. Inokuma H, Okuda M, Ohno K, et al (2002): Analysis of the 18S rRNA gene sequence of a Hepatozoon detected in two Japanese dogs. Vet Parasitol, 106, 265-271.
17. Karagenc TI, Pasa S, Kirli G, et al (2006): A parasitological, molecular and serological survey of Hepatozoon canis infection in dogs around the Aegean coast of Turkey. Vet Parasitol, 135, 113-119.
18. Kimura M (1980): A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol, 16, 111-20.
19. Kumar S, Stecher G, Li M, et al (2018): MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Mol Biol Evol, 35, 1547–1549.
20. Majlathova V, Hurnikova Z, Majlath I, et al (2007): Hepatozoon canis infection in Slovakia: imported or autochthonous? Vector Borne Zoonotic Dis, 7, 199-202.
21. Matsuu A, Ono S, Ikadai H, et al (2005): Development of a SYBR green real-time polymerase chain reaction assay for quantitative detection of Babesia gibsoni (Asian genotype) DNA. Journal Veterinary Diagn Invest, 17, 569-573. 22. Najm NA, Meyer-Kayser E, Hoffmann L, et al (2014): Hepatozoon canis in German red foxes (Vulpes vulpes) and their ticks: molecular characterization and the phylogenetic relationship to other Hepatozoon spp. Parasitol Res, 113, 2679-2685. 23. Ogden NH, Lindsay LR (2016): Effects of Climate and Climate Change on Vectors and Vector-Borne Diseases: Ticks Are Different. Trends Parasitol, 32, 646–656.
24. Orkun Ö, Koç N, Sürsal N, et al (2018): Molecular characterization of tick-borne blood protozoa in stray dogs from Central Anatolia Region of Turkey with a high-rate Hepatozoon infection. Kafkas Univ Vet Fak Derg, 24, 227-232, 2018.
25. Orkun Ö, Nalbantoğlu S (2018): Hepatozoon canis in Turkish red foxes and their ticks. Vet Parasitol Reg Stud Rep, 13, 35–37.
26. Otranto D, Dantas-Torres F, Breitschwerdt EB (2009): Managing canine vector-borne diseases of zoonotic concern: Part two. Trends Parasitol, 25, 228–235.
27. Otranto D, Dantas-Torres F, Weigl S, et al (2011): Diagnosis of Hepatozoon canis in young dogs by cytology and PCR. Parasit Vectors 4, 55.
28. Rochlin I, Toledo A (2020): Emerging tick-borne pathogens of public health importance: a mini-review. J Med Microbiol, 69, 781-791.
29. Stich RW, Blagburn BL, Bowman DD, et al (2014): Quantitative factors proposed to influence the prevalence of canine tick-borne disease agents in the United States. Parasit Vectors, 7, 417.
30. Sutherst RW (2004): Global change and human vulnerability to vectorborne diseases. Clin Microbiol Rev, 17, 136–173.
31. Tuna GE, Bakirci S, Ulutaş B (2020): Evaluation of clinical and haematological findings of mono-and co-infection with Hepatozoon canis in dogs. Animal Health Prod Hyg, 9, 696-702
32. Tuzdil AN (1933): Bizde ilk defa görülen bir Hepatozoon canis vakası. Türk Bay Cem Mec, 13, 35.
33. Ul-Hasan M, Abubakar M, Muhammad G, et al (2012): Prevalence of tick infestation (Rhipicephalus sanguineus and Hyalomma anatolicum anatolicum) in dogs in Punjab, Pakistan. Vet Italia, 48, 95–98
34. Vincent-Johnson NA, Macintire DK, Lindsay DL, et al (1997): A new Hepatozoon species from dogs: description of the causative agent of canine hepatozoonosis in North America. J Parasitol, 83, 1165–1172. 35. Vojta L, Mrljak V, Ćurković S, et al (2009): Molecular epizootiology of canine hepatozoonosis in Croatia. Int J Parasitol, 39, 1129–1136 36. Voyvoda H, Pasa S, Uner A (2004): Clinical Hepatozoon canis infection in a dog in Turkey. J Small Anim Pract, 45, 613-61.

There are 32 citations in total.

Details

Primary Language	English
Subjects	Veterinary Parasitology
Journal Section	Research Article
Authors	Metin Pekağırbaş 0000-0003-3170-410X Muhammed Veli Demirbilek 0009-0002-0173-7170 Emrah Simsek 0000-0002-0492-9840 Heycan Berk Aydın 0009-0007-9466-3487 Hakan Kanlıoğlu 0000-0003-4949-944X Asude Gülçe Oryaşın 0000-0002-7219-2879 Nuran Selek 0000-0001-6223-058X
Early Pub Date	February 14, 2025
Publication Date
Submission Date	October 9, 2024
Acceptance Date	December 8, 2024
Published in Issue	Year 2025Accepted Papers

Cite

APA	Pekağırbaş, M., Demirbilek, M. V., Simsek, E., Aydın, H. B., et al. (2025). Molecular characterization of Hepatozoon spp. in naturally infected dogs in Aydın province. Ankara Üniversitesi Veteriner Fakültesi Dergisi1-6. https://doi.org/10.33988/auvfd.1561161
AMA	Pekağırbaş M, Demirbilek MV, Simsek E, Aydın HB, Kanlıoğlu H, Oryaşın AG, Selek N. Molecular characterization of Hepatozoon spp. in naturally infected dogs in Aydın province. Ankara Univ Vet Fak Derg. Published online February 1, 2025:1-6. doi:10.33988/auvfd.1561161
Chicago	Pekağırbaş, Metin, Muhammed Veli Demirbilek, Emrah Simsek, Heycan Berk Aydın, Hakan Kanlıoğlu, Asude Gülçe Oryaşın, and Nuran Selek. “Molecular Characterization of Hepatozoon Spp. In Naturally Infected Dogs in Aydın Province”. Ankara Üniversitesi Veteriner Fakültesi Dergisi, February (February 2025), 1-6. https://doi.org/10.33988/auvfd.1561161.
EndNote	Pekağırbaş M, Demirbilek MV, Simsek E, Aydın HB, Kanlıoğlu H, Oryaşın AG, Selek N (February 1, 2025) Molecular characterization of Hepatozoon spp. in naturally infected dogs in Aydın province. Ankara Üniversitesi Veteriner Fakültesi Dergisi 1–6.
IEEE	M. Pekağırbaş, M. V. Demirbilek, E. Simsek, H. B. Aydın, H. Kanlıoğlu, A. G. Oryaşın, and N. Selek, “Molecular characterization of Hepatozoon spp. in naturally infected dogs in Aydın province”, Ankara Univ Vet Fak Derg, pp. 1–6, February 2025, doi: 10.33988/auvfd.1561161.
ISNAD	Pekağırbaş, Metin et al. “Molecular Characterization of Hepatozoon Spp. In Naturally Infected Dogs in Aydın Province”. Ankara Üniversitesi Veteriner Fakültesi Dergisi. February 2025. 1-6. https://doi.org/10.33988/auvfd.1561161.
JAMA	Pekağırbaş M, Demirbilek MV, Simsek E, Aydın HB, Kanlıoğlu H, Oryaşın AG, Selek N. Molecular characterization of Hepatozoon spp. in naturally infected dogs in Aydın province. Ankara Univ Vet Fak Derg. 2025;:1–6.
MLA	Pekağırbaş, Metin et al. “Molecular Characterization of Hepatozoon Spp. In Naturally Infected Dogs in Aydın Province”. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 2025, pp. 1-6, doi:10.33988/auvfd.1561161.
Vancouver	Pekağırbaş M, Demirbilek MV, Simsek E, Aydın HB, Kanlıoğlu H, Oryaşın AG, Selek N. Molecular characterization of Hepatozoon spp. in naturally infected dogs in Aydın province. Ankara Univ Vet Fak Derg. 2025:1-6.

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