Molecular and Microscopic Detection of Theileria spp. among Cattle and Buffaloes in West Azarbaijan, Iran

Document Type: Short Communication

Authors

1 Department of Pathobiology, Faculty of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Parasitology and Mycology, Faculty of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran

3 Department of Pathobiology, Faculty of Veterinary Science, Tehran University, Tehran, Iran

4 Department of Microbiology, Faculty of Specialized Veterinary Science, Branch of Science and Research, Islamic Azad University, Tehran, Iran

Abstract

Bovine theileriosis is an important tick-borne disease caused by intraerythrocytic parasites from genus Theileria. This study sought to detect the theileriosis among cattle and buffaloes using molecular and microscopic tests in West Azerbaijan, Iran. For this purpose, 484 blood samples from 193 cattle and 291 buffaloes were collected during March to July 2014. The breed, gender, age, and habitat of these animals were recorded. These animals were native and apparently healthy, living in four different cities of the province. The blood films were stained with Giemsa’s for microscopic examinations. Direct cell semi-nested polymerase chain reaction (PCR) assay was performed to detect T.annulata DNA with Tbs-S/Tbs-A and To-S/Tbs-A primer pairs targeted to 18S ribosomal RNA gene for Theileria spp. and T.orientalis amplification, respectively. The molecular assays revealed that 36 cattle (18.65%) were infected, in which 15 cattle were infected by both T.annulata and T.orientalis. Out of 291 buffaloes, four samples (1.4%) were infected by Theileria genotypes, and two buffaloes (0.7%) were infected only by T.orientalis. The observational results of the gender, age, and habitat of the studied animals were similar to animals of the other parts of Iran. The present study indicated that T.orientalis may be prevalent in native cattle and buffaloes throughout the northern parts of Iran. This study assessed the infection of buffaloes with T.orientalis for the first time.

Keywords

Main Subjects


Article Title [French]

Diagnostic moléculaire et microscopique de Theileria orientalis chez les vaches de la province de l’Azerbaïdjan occidental, Iran

Abstract [French]

La theilériose bovine est une maladie importante causée par une cellule intra-érythrocytaire de Theileria. L’objectif de cette étude est de déterminer à l’aide de méthodes moléculaires et microscopiques l'état actuel de l’infection de Theileria chez les vaches de la province de l'Azerbaïdjan occidental en Iran. Dans cette étude, 484 échantillons de sang ont été prélevés entre mars et juillet 2014 à partir de vaches et buffles indigènes provenant de quatre villes différentes. Les animaux étudiés avaient été classés selon la race, le sexe, l’âge et la ville. Tous les échantillons sanguins des animaux étudués ont fait l’objet d’une coloration de Giemsa pour les observations parasitologiques . Les techniques de PCR et semi-nested PCR ont été également utilisées pour détecter l'ADN de Theileria orientalis à l’aide des paires d’ARN ribosomiques 18S: les ensemble d’amorces Tbs-S/Tbs-A et To/Tbs-A ont été respectivement utilisés pour l’amplification de Theileiria, et Theileria orientalis. . Les analyses moléculaires sur 484 échantillons ont détecté 40 échantillons infectés par le Theileria (8,26%) et 17 (3,51%) échantillons positifs pour le Theileria orientalis. La prévalence de Theileria orientalis chez les vaches indigènes et les buffles était respectivement de 8.33% et 0.68%, révélant pour la première fois une différence significative entre ces deux groupes (p<0.05). L’utilisation de cette approche moléculaire a confirmé l’existence d’infection au Theileria orientalis chez les vaches indigènes ainsi que chez certains buffles originaires plus particulièrement du nord-ouest de l'Iran.

Keywords [French]

  • Theileria orientalis
  • buffles
  • vaches
  • réaction en chaîne de polymérase
  • Iran
Ahmed, J.S., Luo, J., Schnittger, L., Seitzer, U., Jongejan, F., Yin, H., 2006. Phylogenetic position of small-ruminant infecting piroplasms. Ann N Y Acad Sci 1081, 498-504.

Akbari, j., Javanbakht, J., Tavassoli, M., Tabatabai, M., Shafiei, R., 2012. Molecular survey of Theileria annulata in cattle by PCR-RFLP method in Iran. Bacteriol Parasitol 3, 1-6.

Aktas, M., Altay, K., Dumanli, N., 2006. A molecular survey of bovine Theileria parasites among apparently healthy cattle and with a note on the distribution of ticks in eastern Turkey. Vet Parasitol 138, 179-185.

Almeria, S., Castellà, J., Ferrer, D., Ortuño, A., Estrada-Peña, A., Gutiérrez, J.F., 2001. Bovine piroplasms in Minorca (Balearic Islands, Spain): a comparison of PCR-based and light microscopy detection. Veterinary Parasitology 99, 249-259.

Altay, K., Aydin, M.F., Dumanli, N., Aktas, M., 2008. Molecular detection of Theileria and Babesia infections in cattle. Vet Parasitol 158, 295-301.

Dumanli, N., Aktas, M., Cetinkaya, B., Cakmak, A., Koroglu, E., Saki, C.E., et al., 2005. Prevalence and distribution of tropical theileriosis in eastern Turkey. Vet Parasitol 127, 9-15.

Garcı́a-Garcı́a, J.C., Montero, C., Redondo, M., Vargas, M., Canales, M., Boue, O., et al., 2000. Control of ticks resistant to immunization with Bm86 in cattle vaccinated with the recombinant antigen Bm95 isolated from the cattle tick, Boophilus microplus. Vaccine 18, 2275-2287.

Ghaemi, P., Hoghooghi-Rad, N., Shayan, P., Eckert, B., 2012. Detection of Theileria orientalis in Iran by semi-nested PCR. Parasitol Res 110, 527-531.

Hashemi-Fesharki, R., 1988. Control of Theileria annulata in Iran. Parasitology Today 4, 36-40.

Hashemi-Fesharki, R., Habibi, G.R., Ahourai, P., 1998. Delayed type hypersensitivity theilerin test in cattle vaccinated against Theileria annulata infection. Veterinary Parasitology 75, 261-263.

Hayashida, K., Hara, Y., Abe, T., Yamasaki, C., Toyoda, A., Kosuge, T., et al., 2012. Comparative genome analysis of three eukaryotic parasites with differing abilities to transform leukocytes reveals key mediators of Theileria-induced leukocyte transformation. MBio 3, e00204-00212.

Hoghooghi-Rad, N., Ghaemi, P., Shayan, P., Eckert, B., Sadr-Shirazi, N., 2011. Detection of native carrier cattle infected with Theileria annulata by semi-nested PCR in Golestan province of Iran. World Appl Sci J 12 317-323.

Mehlhorn, H., 2008. Encyclopedia of Parasitology: A-M, Springer.

Noaman, V., 2013. A molecular study on Theileria and Babesia in cattle from Isfahan province, Central Iran. J Parasit Dis 37, 208-210.

Ota, N., Mizuno, D., Kuboki, N., Igarashi, I., Nakamura, Y., Yamashina, H., et al., 2009. Epidemiological survey of Theileria orientalis infection in grazing cattle in the eastern part of Hokkaido, Japan. J Vet Med Sci 71, 937-944.

Rahbari, S., Nabian, S., Shayan, P., 2007. Primary report on distribution of tick fauna in Iran. Parasitol Res 101 Suppl 2, S175-177.

Rodriguez-Valle, M., Taoufik, A., Valdes, M., Montero, C., Ibrahin, H., Hassan, S.M., et al., 2012. Efficacy of Rhipicephalus (Boophilus) microplus Bm86 against Hyalomma dromedarii and Amblyomma cajennense tick infestations in camels and cattle. Vaccine 30, 3453-3458.

Safarpour Dehkordi, F., Parsaei, P., Saberian, S., Moshkelani, S., Hajshafiei, P., Hoseini, S.R., 2012. Prevalence study of T.annulata by comparison of four diagnostic techniques in South-west Iran. Bulg J Vet 2, 123-130.

Savini, G., Onuma, M., Scaramozzino, P., Kakuda, T., Semproni, G., Langella, V., 1998. First report of Theileria sergenti and T. buffeli/orientalis in cattle in Italy. Ann N Y Acad Sci 849, 404-407.

Shayan, P., Jafari, S., Fattahi, R., Ebrahimzade, E., Amininia, N., Changizi, E., 2016. Identification and characterization of Theileria ovis surface protein (ToSp) resembled TaSp in Theileria annulata. Parasitol Res 115, 1893-1899.

Shayan, P., Rahbari, S., 2005. Simultaneous differentiation between Theileria spp. and Babesia spp. on stained blood smear using PCR. Parasitol Res 97, 281-286.

Uilenberg, G., 1981. Theilerial Species of Domestic Livestock. In: Irvin, A.D., Cunningham, M.P., Young, A.S. (Eds.), Advances in the Control of Theileriosis: Proceedings of an International Conference held at the International Laboratory for Research on Animal Diseases in Nairobi, 9–13th February, 1981, Springer Netherlands, Dordrecht, pp. 4-37.

Uilenberg, G., Hashemi-Fesharki, R., 1984. Theileria orientalis in Iran. Vet Q 6, 1-4.

Willadsen, P., Smith, D.O.N., Cobon, G., McKenna, R.V., 1996. Comparative vaccination of cattle against Boophilus microplus with recombinant antigen Bm86 alone or in combination with recombinant Bm91. Parasite  Immunology 8, 241-246.

Yokoyama, N., Ueno, A., Mizuno, D., Kuboki, N., Khukhuu, A., Igarashi, I., et al., 2011. Genotypic diversity of Theileria orientalis detected from cattle grazing in Kumamoto and Okinawa prefectures of Japan. J Vet Med Sci 73, 305-312.