A Molecular Study on Hepatozoon canis Infection in Dogs in Tehran (Iran)

Document Type: Original Articles

Authors

1 Department of Microbiology, Faculty of Food Industry and Agriculture, Standard Research Institute (SRI), Karaj, Iran

2 Department of Parasitology and Entomology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

Abstract

Hepatozoonosis is a protozoal disease caused by various species of Hepatozoon. This parasite is transmitted from tick; the main vector of Hepatozoon canis is usually the brown dog tick (Rhipicephalus sanguineus). However, several species of ticks are disposed as the alternative vectors. Dogs are usually infected by eating the tick or a part of the tick organ infected by the mature oocysts containing infectious sporozoite. In the current study, a total of 145 blood samples were collected from the cephalic vein of pet, stray, and shelter dogs in Tehran. To conduct this study, first thin blood smears were prepared from all the samples and stained with the Giemsa method. Then, after extraction of DNA from the blood samples, in order to trace Hepatozoon canis, the 18S rRNA gene segment of the parasite was amplified using polymerase chain reaction (PCR). To confirm the PCR-positive results, five randomly selected PCR-positive samples were sequenced. According to the results, through direct observation of microscopic slides, no infection of H. canis parasite was observed, but according to the PCR results, 32 out of the 145 blood samples were found to be infected by H. canis.  In this study, infection to H. canis in older dogs was higher than in young dogs, and more male dogs were found to be infected by the parasite compared to female dogs; but no significant difference was observed in this regard (P > 0.05). Moreover, stray dogs showed a significantly higher rate of infection, compared to the pet and shelter ones (P < 0.05).

Keywords

Main Subjects


Article Title [French]

Une Étude Moléculaire sur l'Infection à l'Hepatozoon canis chez les Chiens de Téhéran (Iran)

Abstract [French]

L'hépatozoonose est une maladie à protozoaires causée par diverses espèces d'Hepatozoon. Ce parasite est transmis par des tiques et le principal vecteur d'Hepatozoon canis est généralement la tique du chien brun (Rhipicephalus sanguineus). Cependant, plusieurs espèces de tiques représentent des vecteurs alternatifs. Les chiens sont généralement infectés après avoir ingérés la tique ou une partie de leur organe infecté par des oocystes matures contenant des sporozoïtes infectieux. Dans cette étude, un total de 145 échantillons de sang a été prélevé à partir de la veine céphalique des chiens de compagnie, des chiens errants et des chiens d’abri dans la ville de Téhéran. Des premiers frottis sanguins ont été préparés à partir de tous les échantillons et colorés avec la méthode Giemsa. Ensuite, dans le but de détecter la présence d’Hepatozoon canis, l'ADN des échantillons de sang a été extraite, et le segment du gène de l'ARNr 18S du parasite a été amplifié en utilisant des amorces de réaction en chaîne par polymérase (PCR) spécifiques. Pour confirmer les résultats positifs de la PCR, cinq échantillons positifs ont été sélectionnés au hasard et séquencés. D’après les résultats obtenus, aucune infection reliée directement au parasite H. canis n’a été observée à partir des lames microscopiques, alors que les résultats de la PCR montraient que 32 des 145 échantillons desang éxaminés étaient positifs. L'infection à H. canis était plus élevée chez les chiens âgés que chez les jeunes chiens. De plus, il a été constaté que le nombre de chiens infectés par le parasite était supérieur à celui des chiennes; sans pour autant que cela soit statistiquement significatif (p> 0,05). Enfin, les chiens errants présentaient logiquement un taux d'infection significativement plus élevé que les chiens de compagnie et les chiens d’abri (P <0,05).

Keywords [French]

  • Hepatozoon canis
  • Chien
  • PCR
  • Téhéran
Aktas, M., Özübek, S., Altay, K., Balkaya, İ., Utuk, AE., Kırbas, A., Şimsek, S., Dumanlı, N., 2015. A molecular and parasitological survey of Hepatozoon canis in domestic dogs in turkey, Veterinary Parasitology 209, 264-267.

Amoli, A.,A.R., Khoshnegah, j., Razmi, G. R. 2012, A Preliminary Parasitological Survey of Hepatozoon Spp. Infection in Dogs in Mashhad, Iran, Iranian J Parasitol 7, 4, 99-103.

Andre, M.R., Adania, C.H., Teixeira, R.H.F., Vargas, G.H., Falcade, M., Sousa, L., Salles, A.R., Allegretti, S.M., Felippe, P.A.N., Machado, R.Z., 2010, Molecular detection of Hepatozoon spp. In Brazilian and exotic wild carnivores, Veterinary Parasitology173, 134-138.

Aydin, M.F., Sevinc, F., Sevinc, M., 2015, Molecular detection and characterization of Hepatozoon spp. in dogs from the central part of Turkey, Ticks and Tick-borne Diseases 6, 388-392.

Baneth, G., Shkap, V. 1996, Hepatozoon canis: The prevalence of antibodies and gametocytes in dog in Israel, Veterinary Research Communications 20, 1, 41-46.

Baneth, G., Aroch, I. 1997, Hepatozoon canis infection in a Litter of Dalmatian dogs, Veterinary Parasitology 70, 1-3, 201-206.

Baneth, G., Weigler, B. 1997, Retrospective case-control study of hepatozoonosis in dog in Israel, Journal if Veterinary Internal Medicine 11, 6, 365-370.

Baneth, G., Mathew, J.S., Shkap, V. 2003, Canine hepatozoonosis two disease syndromes caused by separate Hepatozoon spp, Trends in Parasitology 19, 1, 27-31.

Baneth, G., Harrus, S., Gal, A., Aroch, I. 2015, Canine vector-borne co-infections: Ehrlichia canis and Hepatozoon canis in the same host monocytes, Veterinary Parasitology 208, 30-34.

Cardoso, L.,  Cortes, C.E.C., Eyal, O.,  Reis, A., Lopes, A.P.,  Vila-Viçosa, M.J., Rodrigues, P.A.,  and  Baneth, G. 2014,  Molecular and histopathological detection of Hepatozoon canis in red foxes (Vulpes vulpes) from Portugal, Parasites and Vectors7, 113-118.

Conceicao, F.M., Abranches, P., Janz, J. G. 1988, Hepatozoonosis in Foxes from Portugal, Journal of Wild life Diseases 24, 2, 344-347.

Criado, F.A., Martinez, M.A., Buling, S.A. 2003, Molecular studies on Babesia, THeileria and Hepatozoon in southern Europ: part I, Veterinary Parasitology 113, 3-4, 189-201.

Dalimi, A., Jameie, F., Mohammadiha, A., Barati, M., Molaei, S. 2017, Molecular detection of Hepatozoon canis in dogs of Ardabil Province, Northwest of Iran. Archiev of Razi Institute 72(3): 197-201.

Dezctek, D., Vojta, L., Curković, S., Lipej, Z., Mihaljević, Z., Cvetnić, Z., Beck, R. 2010, Molecular detection of Theileia annae and Hepatozoon canis in foxes (Vulpes vulpes) in Croatia, Veterinary Parasitology 172, 333-336.

Eiras, F.D., Basabe, J.,Matos, L.M.,Krimer, A.,Baneth, G. 2007, First molecular characterization of canine hepatozoonosis in Argentina: evaluation of asymptomatic Hepatozoon canis infection in dogs from Buenos Aires, Veterinary Parasitology149, 275-279.

Farkas, R., Solymosi, N., Takács., N., Hornyák, A., Hornok, S., Nachum-Biala, Y.,  Baneth, G.2014, First molecular evidence of Hepatozoon canis infection in red foxes and golden jackals from Hungary, Parasites and Vectors7,303.

Gimenez, C., Casado, N.,  Criado-Fornelio, A., de Miguel, F.A., Dominguez-Peñafiel, G. 2009, A molecular survey of Piroplasmida and Hepatozoon isolated from domestic and wild animals in Burgos (northern Spain), Veterinary Parasitology 162, 147-150.

Gomes, P.V.,  Mundim, M.J.S.,  Mundim, A.V., Cury, M.C., 2010, Occurrence of Hepatozoon sp. in dogs in the urbanarea originating from a municipality in southeastern Brazil, Vet. Parasitol 174, 155–161.

Greene, C.E. 2006, Infections Disease of the Dog and Cat, 3rd edition, st, louis: Elsevier.

Inokuma, H., Ohno, K., Yamamoto, S. 1999, Serosurvey of Ehrlichia canis and Hepatozoon canis Infection in Dogs in Yamaguchi Prefecture Japan, The Journal of Veterinary Medical Science 61, 10, 1153-1155.

Kaewkong, W., Intapan, P.M., Sanpool, O.,Janwan, P., Thanchomnang, T., Kongklieng, A., Tantrawatpan, C., Boonmars, T., Lulitanond, V., Taweethavonsawat, P., Chungpivat, S., Maleewong, W., 2014, High throughput pyrosequencing technology for molecular differential detection of Babesia vogeli, Hepatozoon canis, Ehrlichia canis and Anaplasma platys in canine blood samples, Ticks and Tick-borne Diseases 5, 381-385.

Karagenic, I.T., Pasa,S., Kirli, G., Hosgor, M., Bilgic, H.B., Ozon, Y.H., Atasoy, A.,  Eren, H. 2006, A parasitological, molecular and serological survey of Hepatozoon canis infection in dogs around the Aegean coast of Turkey, Veterinary Parasitology 135, 113-119.

Khoshnegah, J., Mohri, M., Movassaghi, A. R., Mehrjerdi, K. H. 2009, The first report of Hepatozoon canis infection of dog in Iran, Comparative Clinical Pathology18, 445-448.

Maede, Y., Ohsagi, T., Ohotaishi, N. 1982, Hepatozoon infection in wild fox (Vulpes vulpes) in Japan, Nippon Juigaku Zasshi44, 1, 137-142.

Macintire, D.K., Vincent-Johnson, N., Dillon, A.R., Blagburn, B., Lindsay, D.,  Whitley, E.M., Banfield, C.1997, Hepatozoonosis in dogs: 22 cases(1989-1994), Journal of the American Veterinary Medical Association 210, 7, 916-922.

Muratu, T., Inoue, M., Tateyama, S., Taura, Y. 1993, Vertical transmission of Hepatozoon canis in dogs, The Journal of Veterinary Medical Science 55, 5, 867-880.

Muratu, T., Inoue, M., Taura, Y. 1995, Detection of Hepatozoon canis oocyst from Ticks collected from the infected dogs, The Journal of Veterinary Medical Science 57, 1, 111-112.

Miranda, de.R.L., O'Dwyer, L.H., Castro, J.R., Metzger, B., Rubini, A.S., Mundim, A.V.,  Eyal, O., Talmi-Frank, D., Cury, M.C., Baneth, G. 2014, Prevalence and molecular characterization of Hepatozoon canis in dogs from urban and rural areas in Southeast Brazil, Research in Veterinary Science 97, 326-329.

Mohanchandra, P.R., Poornachandar, A., Srinivas, P., RamachandraRao, K., Lakshmikantan, U., Shivaji, S. 2012, Molecular characterization of Hepatozoon spp. infectin in endangered Indian wild felids canids, Veterinary Parasitology186, 475, 479.

Otranto, D., Dantas-Torres, F., Weigl, S.,Latrofa, M.S., Stanneck, D., Decaprariis, D., Capelli, G., Baneth, G. 2011, Diagnosis of Hepatozoon canis in young dogs by cytology and PCR. Parasites and Vectors 4, 55.

Rojas, A., Rojas, D., Montenegro, V.M., Baneth, G. 2014, Vector-borne pathogens in dogs from Costa Rica: first molec-ular description of Babesia vogeli and Hepatozoon canis infections witha high prevalence of monocytic ehrlichiosis and the manifestations of co-infection, Vet. Parasitol 199, 121–128.

Sasaki, M., Omobowale, O., Ohta, K., Tozuka, M., Matsuu, A., Hirata, H., Nottidge, H.O., Ikadai, H., Oyamada, T. 2008, A PCR Based Epidemiological Survey of Hepatozoon canis in Dogs in Nigeria, Japanes Society of Veterinary Science 70, 7, 743-745.

Smith, T. G., 1996, The genus Hepatozoon(Apicomplexa: Adeleorina), Journal of Parasitology 82, 4, 565-585.

Waned, T., Baneth, G., Zuckerman., Nyska, A.1994, Hepatozoon canis: size measurment of the gametocyte using image analysis technology, Comparative Haematology International 4, 3, 177-179.

Wall, R., Shearer, D. 2001, Veterinary ectoparasites: biology, pathology and control, ed 2, Blackwell Science: Oxford.