Kursat ALTAY, Mehmet Fatih AYDIN, Ayperi AYTMIRZAKIZI, Zarima JUMAKANOVA, Ayday CUNUSOVA, Nazir DUMANLI

Molecular Survey of Hepatozoonosis in Natural Infected Dogs: First Detection and Molecular Characterisation of Hepatozoon canis in Kyrgyzstan

Canine hepatozoonosis is a tick-borne protozoan disease and widespread in Europe, Africa, Asia and America. There is not any available data about the presence of Hepatozoon infections in dogs in Kyrgyzstan. In the study we aimed that investigate the presence of Hepatozoon canis and the prevalance of Hepatozoon infections in dogs from Kyrgyzstan using polymerase chain reaction (PCR) and sequence analysis. To determine the prevelance of hepatozoonosis in dogs, a total of 170 blood samples were applied to PCR to amplify a fragment of 666 bp found in 18S SSU rRNA gene of Hepatozoon spp. The PCR results shown that Hepatozoon infection rate was 28.8% (49/170) in dogs. Eleven representative positive samples were sequenced to classification of the species. The nucleotide sequences were compared to the H. canis sequences which registered in GenBank using the basic local alignment search tool. Results of sequence analyse of 11 amplicons indicated that 8 were 100% identical and the other 3 sequences shared 99% similarity with H. canis. The sequences were deposited in Genbank with accession numbers from MG917709 to MG917719. It was the first record of H. canis in dogs in Kyrgyzstan.

Doğal Enfekte Köpeklerde Hepatozoon Enfeksiyonlarının Moleküler Yöntemlerle Araştırılması: Hepatozoon canis’in Kırgızistan’da İlk Tespiti

Köpeklerde hepatozoonozis kenelerle nakledilen ve Afrika, Avrupa, Asya ile Amerika’da yaygın olarak görülen protozoan bir hastalıktır. Kırgızistan’da köpeklerde Hepatozoon enfeksiyonlarının varlığına dair bir bilgiye ulaşılamamıştır. Bu çalışmada, polimeraz zincir reaksiyonu (PZR) ve sekans analizi ile Kırgızistan’da köpeklerde Hepatozoon enfeksiyonlarının yaygınlığının ve Hepatozoon canis’in varlığının belirlenmesi amaçlanmıştır. Hepatozoon enfeksiyonlarının prevalansını belirlemek için 170 kan örneğine, Hepatozoon spp. 18S SSU rRNA geninin 666 bp’lik kısımını amplifiye etmek üzere PZR uygulanmıştır. PZR sonucunda Hepatozoon spp. enfeksiyon oranı %28.8 (49/170) olarak ortaya çıkmıştır. On bir PZR pozitif DNA örneğinin DNA dizilimi belirlenmiş ve elde edilen DNA dizilimleri BLAST programı kullanılarak GenBank’ta kayıtlı diğer dizilimlerle karşılaştırılmıştır. Bunlardan 8’inin GenBankasında kayıtlı H. canis DNA dizilimleri ile %100 oranında eşleştiği, diğer 3 dizilimin ise %99 oranında bir benzerliğe sahip olduğu belirlenmiştir. DNA dizilimleri GenBankasına MG917709-MG917719 numaraları altında kaydedilmiştir. Bu çalışma Kırgızistan’da köpeklerde H. canis’in belirlendiği ilk çalışma niteliğindedir.

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1. Baneth G: Perspectives on canine and feline hepatozoonosis. Vet Parasitol, 181, 3-11, 2011. DOI: 10.1016/j.vetpar.2011.04.015

2. Dantas-Tores F, Otranto D: Hepatozoonosis. In, Marcondes CB (Ed): Arthropod Borne Diseases, Springer International Publishing Switzerland, 363-368, 2017. DOI: 10.1007/978-3-319-13884-8

3. Smith TG: The genus Hepatozoon (Apicomplexa: Adeleina). J Parasitol, 82, 565-585, 1996. DOI: 10.2307/3283781

4. Aydin MF, Sevinc F, Sevinc M: Molecular detection and characterization of Hepatozoon spp. in dogs from central part of Turkey. Ticks Tick Borne Dis, 6, 388-392, 2015. DOI: 10.1016/j.ttbdis.2015.03.004

5. Orkun Ö, Koç N, Sürsal N, Çakmak A, Nalbantoğlu S, Karaer Z: 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 (2): 227-232, 2018. DOI: 10.9775/ kvfd.2017.18678

6. Baneth G, Samish M, Alekseev E, Aroch I, Shkap V: Transmission of Hepatozoon canis to dogs by naturally fed or percutaneously injected Rhipicephalus sanguineus ticks. J Parasitol, 87, 606-611, 2001. DOI: 10.1645/0022-3395(2001)087[0606:TOHCTD]2.0.CO;2

7. Baneth G, Samish M, Shkap V: Life cycle of Hepatozoon canis (Apicomplexa: Adeleorina: Hepatozoidae) in the ticks Rhipicephalus sanguineus and domectic dogs (Canis familiaris). J Parasitol, 93 (2): 283- 299, 2007. DOI: 10.1645/GE-494R.1

8. Mathew JS, Ewing SA, Panciera RJ, Woods JP: Experimental transmission of Hepatozoon americanum Vincent- Johnson et al., 1997 to dogs by the Gulf Coast tick, Amblyomma maculatum Koch. Vet Parasitol, 80, 1-14, 1998. DOI: 10.1016/S0304-4017(98)00189-7

9. Taylor MA, Coop RL, Wall RL, Parasites of dogs and cats. Veterinary Parasitology, Blackwell Publishing, 409-433, 2007.

10. Baneth G, Vicent-Jhonson N: Hepatozoonosis. In, Shaw SE, Day MJ (Eds): Arthropod Borne Infectious Diseases of the Dog and Cat. 78-88, Manson Publishing Ltd, London, 2005.

11. Eiras DF, Basabe J, Scodellaro CF, Banach DB, Matos ML, Krimer A, Baneth G: First molecular characterization of canine hepatozoonosis in Argentina: Evaluation of asymptomatic Hepatozoon canis infection in dogs from Buenos Aires. Vet Parasitol, 149, 275-279, 2007. DOI: 10.1016/j.vetpar.2007.07.010

12. Aktas M, Ozubek S, Altay K, Sayın Ipek ND, Balkaya I, Utuk AE, Kirbas A, Simsek S, Dumanli N: Molecular detection of tick-borne rickettsial and protozoan pathogens in domestic dogs from Turkey. Parasit Vectors, 8:157, 2015. DOI: 10.1186/s13071-015-0763-z

13. Aktas M, Ozubek S, Altay K, Balkaya I, Utuk AE, Kirbas A, Simsek S, Dumanli N: A molecular and parasitological survey of Hepatozoon canis in domestic dogs in Turkey. Vet Parasitol, 209, 264-267, 2015. DOI: 10.1016/j.vetpar.2015.02.015

14. Otranto D, Dantas-Torres F, Weigl S, Latrofa MS, Stanneck D, de Caprariis D, Capelli G, Baneth G: Diagnosis of Hepatozoon canis in young dogs by cytology and PCR. Parasit Vectors, 4:55, 2011. DOI: 10.1186/1756- 3305-4-55

15. Altay K, Dumanlı N, Aktaş M, Özübek S: Survey of Anaplasma infections in small ruminants from East part of Turkey. Kafkas Univ Vet Fak Derg, 20 (1): 1-4, 2014. DOI: 10.9775/kvfd.2013.9189

16. Altay K, Ataş AD, Özkan E: Sivas yöresi koyun keçi ve kenelerde Theileria ve Babesia türlerinin moleküler yöntemlerle araştırılması. Manas J Agr Vet Life Sci, 7 (1): 30-39, 2017.

17. Altay K, Dumanlı N, Holman PJ, Aktaş M: Detection of Theileria ovis in naturally infected sheep by nested PCR. Vet Parasitol, 127 (2): 99-104, 2005. DOI: 10.1016/j.vetpar.2004.09.012

18. Altay K, Aktaş M, Dumanlı N: Doğu ve Güneydoğu Anadolu Bölgelerinde küçük ruminantlarda Theileria enfeksiyonları. Türkiye Parazitol Derg, 31 (4): 268-271, 2007.

19. Dumanlı N, Aktaş M, Çetinkaya B, Çakmak A, Köroğlu E, Saki CE, Erdoğmuş Z, Nalbantoğlu S, Öngör H, Şimşek S, Karahan M, Altay K: Prevalence and distribution of tropical theileriosis in eastern Turkey. Vet Parasitol, 127 (1): 9-15, 2005. DOI: 10.1016/j.vetpar.2004.08.006

20. Karagenc TI, Pasa S, Kirli G, Hosgor M, Bilgic HB, Ozon YH, Atasoy A, Eren H: A parasitological, molecular and serological survey of Hepatozoon canis infection in dogs around the Aegean coast of Turkey. Vet Parasitol, 135, 113-119, 2006. DOI: 10.1016/j.vetpar.2005.08.007

21. Pasa S, Kiral F, Karagenc T, Atasoy A, Seyrek K: Description of dogs naturally infected with Hepatozoon canis in the Aegean region of Turkey. Turk J Vet Anim Sci, 33 (4): 287 289-295, 2009. DOI: 10.3906/vet-0801-11

22. Altay K, Dumanlı N, Aktaş M: A study on ovine tick-borne hemoprotozoan parasites (Theileria and Babesia) in the East Black Sea Region of Turkey. Parasitol Res, 111 (1): 149-153, 2012. DOI: 10.1007/ s00436-011-2811-8

23. Ilyasov S, Zabenko O, Gaydamak N, Kirilenko A, Myrsaliev N, Shevchenko V, Penkina L: Climate profile of the Kyrgyz Republic, The State Agency for Environmental Protection and Forestry under the Government of the Kyrgyz Republic, The United Nations Development Programme,Bishkek, 2013. http://www.kg.undp.org/content/dam/ kyrgyzstan/Publications/env-energy/kgz_Kyrgyzstan%20Climate%20 profile_ENG_for%20web-opt.pdf; Accessed: 17.05.2018.

24. Inokuma H, Okuda M, Ohno K, Shimoda K, Onishi T: Analysis of the 18S rRNA gene sequence of a Hepatozoon detected in two Japanese dogs. Vet Parasitol, 106, 265-271, 2002. DOI: 10.1016/S0304-4017 (02)00065-1

25. Sasaki M, Omobowale O, Ohta K, Tozuka M, Matsuu A, Hirata H, Nottidge HO, Ikadai H, Oyamada T: A PCR-based epidemiological survey of Hepatozoon canis in dogs in Nigeria. J Vet Med Sci, 70 (7): 743- 745, 2008. DOI: 10.1292/jvms.70.743

26. Jittapalapong S, Rungphisutthipongse O, Maruyama S, Schaefer JJ, Stich RW: Detection of Hepatozoon canis in stray dogs and cats in Bangkok Thailand. Ann N Y Acad Sci, 1081, 479-488, 2006. DOI: 10.1196/ annals.1373.071

27. Vojta L, Mrljak V, Curkovic S, Zivicnjak T, Marinculic A, Beck R: Molecular epizootiology of canine hepatozoonosis in Croatia. Int J Parasitol, 39, 1129-1136, 2009. DOI: 10.1016/j.ijpara.2009.02.007

28. Rubini AS, dos Santos Paduan K, Von Ah Lopes V, O’Dwyer LH: Molecular and parasitological survey of Hepatozoon canis (Apicomplexa: Hepatozoidae) in dogs from rural area of Sao Paulo state, Brazil. Parasitol Res, 102 (5): 895-899, 2008. DOI: 10.1007/s00436-007-0846-7

29. Ivanov A, Tsachev I: Hepatozoon canis and hepatozoonosis in the dog. Trakia J Sci, 6 (2): 27-35, 2008.

30. Penzhorn BL, Netherlands EC, Cook CA, Smit NJ, Vorster I, Harrison- White RF, Oosthuizen MC: Occurrence of Hepatozoon canis (Adeleorina: Hepatozoidae) and Anaplasma spp. (Rickettsiales: Anaplasmataceae) in black-backed jackals (Canis mesomelas) in South Africa. Parasit Vectors, 11:158, 2018. DOI: 10.1186/s13071-018-2714-y