Molecular Detection of Hepatozoon canis in Dogs of Ardabil Province, Northwest of Iran

Document Type: Short Communication

Authors

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

2 Faculty of Medical Sciences, Tarbiat Modares University

3 Infectious diseases Research Center, Aja University of Medical Sciences

Abstract

Hepatozoon species are protozoan parasites that infect some animals such as birds, reptiles, amphibians, and carnivores. Previous studies performed on canine hepatozoonosis in Iran have never used molecular techniques for diagnosis of this disease. The main objective of the present study was to detect Hepatozoon canis in the blood of dogs using polymerase chain reaction (PCR) method and sequencing. A total of 104 blood samples were collected from dogs of Meshginshahr County (Ardabil Province), and DNA was extracted from blood samples by dint of DNG-plus Extraction Kit. Then, 18S rRNA gene was amplified by using the conventional PCR methods. PCR products yielded an amplicon of the approximate length of 897 bp for all the positive samples. Twenty-four out of the 104 (23.07%) samples were found to be positive for H. canis. This rate of infection is relatively high among dogs in Ardabil Province. Sequence analysis confirmed the molecular identity of 99% of the samples by comparison with GenBank profiles. This is the first report of molecular detection of H. canis from Iran.

Keywords

Main Subjects


Article Title [French]

Diagnostic moléculaire de l’hépatozoonose canine dans la province d’Ardabil, nord-ouest de l'Iran

Abstract [French]

Les espèces d’hépatozoonose sont des protozoaires infectant les animaux tels que les oiseaux, les reptiles, les amphibiens et les carnivores. Bien que l’hépatozoonose canine ait été rapportée en Iran, elle n'a pas encore été détectée par des méthodes moléculaires. L'objectif principal de cette étude était le diagnostic moléculaire de l’hépatozoonose canine par la méthode PCR et la détermination de sa séquence nucléotidique. A cet effet, 104 échantillons de sang provenant des chiens de la ville de Meshkinshahr (Province Ardabil) ont été recueillies et les ADNs ont été extraites à l’aide du kit DNG-PLUS. Ensuite le gène SrRNA 18 a été amplifié par PCR conventionnelle afin de déterminer sa séquence. Une bande d'environ 897 pb a été identifiée dans les échantillons positifs et 24 des 104 échantillons analysés (23.7%) étaient infectés par une hépatozoonose canine. Ce pourcentage d'infestation chez les chiens de la province Ardabil est relativement élevé. La séquence moléculaire du fragment étudié était 99% similaire à celles enregistrées dans la banque de gènes. Cette étude représente le premier rapport sur la détection de l’hépatozoonose canine en Iran.

Keywords [French]

  • Hépatozoonose canine
  • PCR
  • Chien
  • Province d’Ardabil
  • Iran
Aktas, M., Ozubek, S., Altay, K., Balkaya, I., Utuk, A.E., Kirbas, A., et al., 2015. A molecular and parasitological survey of Hepatozoon canis in domestic dogs in Turkey. Vet Parasitol 209, 264-267.

Aktas, M., Ozubek, S., Ipek, D.N., 2013. Molecular investigations of Hepatozoon species in dogs and developmental stages of Rhipicephalus sanguineus. Parasitol Res 112, 2381-2385.

Baneth, G., Barta, J.R., Shkap, V., Martin, D.S., Macintire, D.K., Vincent-Johnson, N., 2000. Genetic and antigenic evidence supports the separation of Hepatozoon canis and Hepatozoon americanum at the species level. J Clin Microbiol 38, 1298-1301.

Baneth, G., Samish, M., Alekseev, E., Aroch, I., Shkap, V., 2001a. Transmission of Hepatozoon canis to Dogs by Naturally-Fed or Percutaneously-Injected Rhipicephalus sanguineus Ticks. J Parasitol 87, 606-611.

Baneth, G., Samish, M., Shkap, V., 2007. Life cycle of Hepatozoon canis (Apicomplexa: Adeleorina: Hepatozoidae) in the tick Rhipicephalus sanguineus and domestic dog (Canis familiaris). J Parasitol 93, 283-299.

Baneth, G.A.D., Samish, M., Alekseev, E., Aroch, I., Shkap, V., 2001b. Transmission of Hepatozoon canis to Dogs by Naturally-Fed or Percutaneously-Injected Rhipicephalus sanguineus Ticks.  J Parasitol 87, 606-611.

Christophers, S.R., 2016. Leucocytozoon Canis (Classic Reprint), Fb &C Limited.

Criado-Fornelio, A., Martinez-Marcos, A., Buling-Saraña, A., Barba-Carretero, J.C., 2003. Molecular studies on Babesia, Theileria and Hepatozoon in southern Europe. Vet Parasitol 114, 173-194.

Criado-Fornelio, A., Rey-Valeiron, C., Buling, A., Barba-Carretero, J.C., Jefferies, R., Irwin, P., 2007. New advances in molecular epizootiology of canine hematic protozoa from Venezuela, Thailand and Spain. Vet Parasitol 144, 261-269.

Dissanaike, A.S., 1961. Hepatozoon canis infection in dogs in Ceylon. Ceylon Vet J 9, 144–145.

Elias, E., Homans, P.A., 1988. Hepatozoon canis infection in dogs: clinical and haematological findings; treatment. Journal of Small Anim Pract 29, 55-62.

Gomes, P.V., Mundim, M.J., Mundim, A.V., de Avila, D.F., Guimaraes, E.C., Cury, M.C., 2010. Occurrence of Hepatozoon sp. in dogs in the urban area originating from a municipality in southeastern Brazil. Vet Parasitol 174, 155-161.

Gondim, L.s.F.P., Kohayagawa, A., Alencar, N.X., Biondo, A.W., Takahira, R.K., Franco, S.R.V., 1998. Canine hepatozoonosis in Brazil: description of eight naturally occurring cases. Vet Parasitol 74, 319-323.

Gonen, L., Strauss-Ayali, D., Shkap, V., Vincent-Johnson, N., Macintire, D.K., Baneth, G., 2004. An enzyme-linked immunosorbent assay for antibodies to Hepatozoon canis. Vet Parasitol 122, 131-139.

Inokuma, H., Ohno, K., Yamamoto, S., 1999. Serosurvey of Ehrlichia canis and Hepatozoon canis infection in dogs in Yamaguchi Prefecture, Japan. J Vet Med Sci 61, 1153-1155.

Inokuma, H., Okuda, M., Ohno, K., Shimoda, K., Onishi, T., 2002. Analysis of the 18S rRNA gene sequence of a Hepatozoon detected in two Japanese dogs. Veterinary Parasitology 106, 265-271.

James, S.P., 1905. On a parasite found in the white corpuscles of the blood of dogs. Sci Mem Offrs Med Sanit Deps India 14, 1-12.

Khazeni, A., Telmadarraiy, Z., Oshaghi, A., Mohebali, M., Zarei, Z., Abtahi, S.M., 2013. Molecular detection of <i>Ehrlichia canis</i> in ticks population collected on dogs in Meshkin-Shahr, Ardebil Province, Iran. J Biomed Sci Engin 06, 5.

Khoshnegah, J., Mohri, M., Movassaghi, A.R., Mehrjerdi , H.K., 2009. The first report of Hepatozoon canis infection of a dog in Iran. Com Clin Pathol 18, 455-458.

Murata, T., Shiramizu, K., Hara, Y., Inoue, M., Shimoda, K., Nakama, S., 1991. First case of Hepatozoon canis infection of a dog in Japan. J Vet Med Sci 53, 1097-1099.

O’Dwyer, L.H., Massard, C.L., Pereira de Souza, J.C., 2001.Hepatozoon canis infection associated with dog ticks of rural areas of Rio de Janeiro State, Brazil. Veterinary Parasitology 94, 143-150.

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

Paludo, G.R., Dell’Porto, A., de Castro e Trindade, A.R., McManus, C., Friedman, H., 2003. Hepatozoon spp.: report of some cases in dogs in Brası́lia, Brazil. Vet Parasitol 118, 243-248.

Pawar, R.M., Poornachandar, A., Srinivas, P., Rao, K.R., Lakshmikantan, U., Shivaji, S., 2012. Molecular characterization of Hepatozoon spp. infection in endangered Indian wild felids and canids. Vet Parasitol 186, 475-479.

Rahmani Amoli, A.A., Khoshnegah, J., Razmi, G.R., 2012. A preliminary parasitological survey of Hepatozoon spp. infection in dogs in Mashhad, Iran. Iranian J Parasitol 7, 99-103.

Rajamanickam, C., Wiesenhutter, E., Zin, F.M., Hamid, J., 1985. The incidence of canine haematozoa in peninsular Malaysia. Vet Parasitol 17, 151-157.

Razmi, G.R., Naghibi, A., Aslani, M.R., Dastjerdi, K., Hossieni, H., An epidemiological study on Babesia infection in small ruminants in Mashhad suburb, Khorasan province, Iran. Small Rumin Res 50, 39-44.

Rubini, A.S., dos Santos Paduan, K., Cavalcante, G.G., Ribolla, P.E., O'Dwyer, L.H., 2005. Molecular identification and characterization of canine Hepatozoon species from Brazil. Parasitol Res 97, 91-93.

Shkap, V., Baneth, G., Pipano, E., 1994. Circulating antibodies to Hepatozoon canis demonstrated by immunofluorescence. J Vet Diagn Invest 6, 121-123.

Vargas-Hernandez, G., Andre, M.R., Faria, J.L., Munhoz, T.D., Hernandez-Rodriguez, M., Machado, R.Z., et al., 2012. Molecular and serological detection of Ehrlichia canis and Babesia vogeli in dogs in Colombia. Vet Parasitol 186, 254-260.