Original Scientific Article
Molecular detection and phylogenetic analysis of Anaplasma spp. in cattle in Al-Qadisiyah province of Iraq
Correspondence: Yahia I. Khudhair, firstname.lastname@example.org
Received: 16 January 2019
Received in revised form: 17 July 2019
Accepted: 20 August 2019
Available Online First: 10 September 2019
Published on: 15 October 2019
Anaplasma spp. are widely spread rickettsial bacteria transmitted by ticks and placing high impacts on veterinary and public health. A limited number of studies have been carried out on Anaplasmosis in the central part of Iraq. This study was conducted to determine the presence of Anaplasma spp. in cattle in Al-Qadisiyah province, Iraq. A total of 400 blood specimens were collected from cattle suffering from heavy tick infestation. Cattle were blood-sampled from four hyperendemic areas with ticks. Blood samples were screened using microscopic and polymerase chain reaction (PCR) methods. Diff-quick stained blood smears revealed Anaplasma-like inclusion bodies in 254 (63.5%) samples. According to the 16S rRNA-gene-based PCR analysis, Anaplasma spp. was detected in 124 of the 400 (31%) samples, divided as 96/254 (37.8%) among the microscopical positive samples and 28/146 (19.17%) among the microscopical negative samples. Phylogenetic analysis based on the partial 16S rRNA gene sequencing of ten-PCR positive samples were 99–97% identical to sequences deposited in the GenBank, revealing presence of A. phagocytophilum, A. marginale and unnamed Anaplasma spp. in 40%, 20%, and 40% samples, respectively. Relationships among Anaplasma spp. infections and cattle breed, age, and sex were analyzed. Calves less than one year old showed significantly higher rates (p<0.005) than those from other age groups, whereas sex and breed demonstrated no significant differences (p˃0.001). This study shows that a variety of Anaplasma spp., were endemic in central part of Iraq and is still a hidden problem in cattle in the hyperendemic areas of tick, which requires serious control strategies.
Keywords: Anaplasma phagocytophilum, bovine anaplasmosis, Iraq, phylogenetic analysis
1. Noaman, V., Shayan, P. (2010). Molecular detection of Anaplasma bovis in cattle from central part of Iran. Veterinary Research Forum. Faculty of Veterinary Medicine, Urmia University 1(2):117-122.
2. Tay, S.T., Koh, F.X., Kho, K.L., Ong, B.L. (2014). Molecular survey and sequence analysis of Anaplasma spp. in cattle and ticks in a Malaysian farm. Tropical biomedicine, 31(4): 769-776.3. Albadrani, B.A., Al-Iraqi, O.M. (2019). First detection of equine anaplasmosis and hemoplasmosis of horses in Mosul city, Iraq. Adv. Anim. Vet. Sci. 7(2): 106-111.4. Zobba, R., Anfossi, A.G., Parpaglia, M.L.P., Dore, G.M., Chessa, B., et al. (2014). Molecular investigation and phylogeny of anaplasma spp. in mediterranean ruminants reveal the presence of neutrophil-tropic strains closely related to A. platys. Appl. Environ. Microbiol. 80(1): 271-280. https://doi.org/10.1128/AEM.03129-13 PMid:24162569 PMCid:PMC39110105. Kocan, K.M., de la Fuente, J., Blouin, E.F., Coetzee, J.F., Ewing, S.A. (2010). The natural history of Anaplasma marginale. Vet. Parasitol. 167(2-4): 95-107.
6. Pazhoom, F., Ebrahimzade, E., Shayan, P., Nabian, S. (2016). Anaplasma spp. identification in hard ticks of Iran: First report of Anaplasma bovis in Haemaphysalis inermis. Acarologia 56(4): 497-504. https://doi.org/10.1051/acarologia/201641377. Yang, J., Liu, Z., Niu, Q., Luo, J., Wang, X., Yin, H. (2017). Molecular detection of Anaplasma phagocytophilum in wild cervids and hares in China. J. Wildl. Dis. 53(2): 420-423. https://doi.org/10.7589/2016-09-215 PMid:281510848. Dondi, F., Russo, S., Agnoli, C., Mengoli, N., Balboni, A., et al. (2014). Clinicopathological and molecular findings in a case of canine Anaplasma phagocytophilum infection in Northern Italy. Sci. World J. 2014: 810587. https://doi.org/10.1155/2014/810587 PMid:25003154 PMCid:PMC40705429. Khatat, S.E.H., Sahibi, H. (2015). Anaplasma phagocytophilum: An emerging but unrecognized tick-borne pathogen. Rev. Mar. Sci. Agron. Vét. 3(2): 43-52.10. Awad, A.H.H., Abdul-Hussein, M.A. (2006). New record of two species of hard ticks from some domestic animals in Basrah-Iraq. Journal of Basrah Researches (Sciences) 32(1A): 1-6.11. Mallah, M.O., Rahif, R.H. (2016). Epidemiological study for ticks infestation in cattle in Baghdad city- Iraq. Iraqi Journal od Veterinary Sciences 15(2): 45-51.12. Werszko, J., Szewczyk, T., Steiner-Bogdaszewska, Ż., Laskowski, Z., Karbowiak, G. (2019). Molecular detection of Anaplasma phagocytophilum in blood- sucking flies (Diptera: Tabanidae) in Poland. J. Med. Entomol. 56(3): 822- 827. https://doi.org/10.1093/jme/tjy217 PMid:3061516813. Masuzawa, T., Uchishima, Y., Fukui, T., Okamoto, Y., Muto, M., et al. (2011). Detection of Anaplasma phagocytophilum from wild boars and deer in Japan. Jpn. J. Infect. Dis. 64(4): 333-336.14. Noaman, V., Shayan, P. (2012). Molecular detection of Anaplasma bovis in cattle from central part of Iran. Vet. Res. Forum 1(2): 117-122.15. Noaman, V., Shayan, P. (2009). Molecular detection of Anaplasma phagocytophilum in carrier cattle of Iran - first documented report. Iran. J. Microbiol. 1(2): 37-42.16. Koval'Chuk, S.N., Kosovskii, G.Y., Arkhipov, A.V., Glazko, T.T., Glazko, V.I. (2015). Development of real-time PCR assay for detection of Anaplasma marginale. Agricultural Biology 50(6): 825-831. https://doi.org/10.15389/agrobiology.2015.6.825eng 17. Yang, J., Han, R., Niu, Q., Liu, Z., Guan, G., et al. (2018). Occurrence of four Anaplasma species with veterinary and public health significance in sheep, northwestern China. Ticks Tick. Borne. Dis. 9(1): 82-85. https://doi.org/10.1016/j.ttbdis.2017.10.005 PMid:2903782618. Khudhair, Y.I. (2018). Molecular survey and phylogeny of Anaplasma ovis in small ruminants in Al-Qadisiyah Province, Iraq. Al-Qadisiyah J. Vet. Med. Sci. 16(2): 137-142.
https://doi.org/10.29079/vol16iss2art45619. Al-gharban, H.A., Dhahir, S.H. (2015). Serological diagnosis of persistent infection with Anaplasma marginale bacteria in cattle. The Iraqi Journal of Veterinary Medicine 39(1): 33-39.20. Brown, G.K., Martin, A.R., Roberts, T.K., Aitken, R.J. (2001). Detection of Ehrlichia platys in dogs in Australia. Australian Veterinary Journal 79(8): 554-558.
https://doi.org/10.1111/j.1751-0813.2001.tb10747.x PMid:1159981721. Mohammad, M.K. (2016). Ixodiod tick fauna infesting sheep and goats in the middle and south of Iraq. Bull. Iraq Nat. Hist. Mus. 14(1): 43-50.22. Mohammad, M.K. (2015). Distribution of ixodid ticks among domestic and wild animals in central Iraq. Bull. Iraq Nat. Hist. Mus. 13(2): 23-30.23. Silaghi, C., Santos, A.S., Gomes, J., Christova, I., Matei, I.A., et al. (2017). Guidelines for the direct detection of Anaplasma spp. in diagnosis and epidemiological studies. Vector-Borne Zoonotic Dis. 17(1): 12-22. https://doi.org/10.1089/vbz.2016.1960 PMid:2805557924. Salehi-Guilandeh, S., Sadeghi-Dehkordi, Z., Sadeghi-Nasab, A., Yousefi, A. (2018). Molecular detection of Anaplasma spp. in cattle of Talesh County, North of Iran. Bulg J Vet Med. Online first. DOI: 10.15547/bjvm.213525. Turi, A.A. (2018). Comparative analysis of indirect ELISA and real time PCR for the detection of anaplasma marginale in buffalo, cattle and sheep in district Peshawar and Lakki Marwat, Pakistan. South Asian J. Life Sci. 6(1): 0-6. https://doi.org/10.17582/journal.sajls/2018/126.96.36.1996. Rahman, W.A., Fong, S., Chandrawathani, P., Nurulaini, R., Zaini, C.M., Premalaatha, B. (2012). Comparative seroprevalences of bovine trypanosomiasis and anaplasmosis in five states of Malaysia. Trop. Biomed. 29(1): 65-70.27. Jassem, G.A., Agaar, O.A. (2015). Molecular and biochemical study of Anaplasma marginale in cattle in Wassit Province of Iraq. African Journal of Bacteriology Research 7(4): 36-41.28. Qiu, H., Kelly, P.J., Zhang, J., Luo, Q., Yang, Y., et al. (2016). Molecular detection of Anaplasma spp. and Ehrlichia spp. in ruminants from twelve provincesof China . Can. J. Infect. Dis. Med. Microbiol. 2016, 1-9. https://doi.org/10.1155/2016/9183861 PMid:28096822 PMCid:PMC520643229. Zhao, S., Cui, Y., Jing, J., Yan, Y., Peng, Y., et al. (2019). Rapid and sensitive detection of Anaplasma phagocytophilum using a newly developed recombinase polymerase amplification assay. Exp. Parasitol. 201, 21-25. https://doi.org/10.1016/j.exppara.2019.04.010 PMid:31029698
© 2019 Ayyez H.N. This is an open-access article published under the terms of the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Conflict of Interest Statement
The authors declared that they have no potential conflict of interest with respect to the authorship and/or publication of this article.
Macedonian Veterinary Review. Volume 42, Issue 2, Pages 181-188, e-ISSN 1857-7415, p-ISSN 1409-7621, DOI: 10.2478/macvetrev-2019-0023, 2019