KEREM ÖTER, HANDAN ÇETİNKAYA, CEM VURUŞANER, MÜFİT TOPARLAK, KORAY ERGÜNAY

Molecular Detection and Typing of Anaplasma Species in Small Ruminants in Thrace Region of Turkey

Bu çalışma, Türkiye'nin Trakya bölgesindeki küçük ruminantlarda Anaplasma ovis ve Anaplasma phagocytophilum'un varlığı ve dağılımını belirlemek amacı ile Mayıs-Eylül 2014 tarihleri arasında İstanbul, Tekirdağ, Edirne ve Kırklareli illerinde yürütülmüştür. Klinik semptom göstermelerine bakılmaksızın rastgele seçilen küçük ruminantlardan toplam 423 kan örneği (216 koyun ve 207 keçi) toplanmıştır. A. ovis ve A. phagocytophilum türlerinin identifikasyonu için major surface protein 4 (msp4) genini hedef alan tür-spesifik polimeraz zincir reaksiyonu (PZR) kullanılmış olup seçilen ürünler sekanslanarak doğrulanmıştır. Toplam 230 (%54.37) küçük ruminantın A. ovis ve/veya A. phagocytophilum ile enfekte olduğu bulunmuştur. A. ovis ve A. phagocytophilum yönünden pozitif hayvanların yüzdesi sırasıyla %50.83 (215/423) ve %8.51 (36/423) bulunmuştur. Her iki tür için pozitif hayvanların yüzdesi %4.96 (21/423) olarak tespit edilmiştir. Sekans farklılıklıkları A. ovis için 0-0.94% ve A. phagocytophilum için 0.41-2.49% oranlarında izlenmiştir. Bu çalışma, A. ovis ve A. phagocytophilum'un Türkiye'nin Trakya bölgesindeki koyun ve keçilerde varlığı ve dağılımı üzerine polimeraz zincir reaksiyonu ve sekans karakterizasyonu ile yapılan ilk araştırmadır. Bölgedeki Anaplasma türlerinin varlık ve dağılımını etkileyebilecek vektör, vektörkonak ilişkileri ve genotipik varyantlar konusunda yeni araştırmalara ihtiyaç duyulmaktadır

Türkiye'nin Trakya Bölgesindeki Küçük Ruminantlarda Görülen Anaplasma Türlerinin Moleküler Yöntemlerle Tespiti ve Tiplendirmesi

This study was conducted to determine the presence and distribution of Anaplasma ovis and Anaplasma phagocytophilum in small ruminants in Istanbul, Tekirdag, Edirne and Kirklareli provinces in Thrace region of northwestern Turkey during May-September in 2014. A total of 423 blood samples (216 sheep and 207 goats) were collected randomly from small ruminants regardless of the clinical symptoms. Species-specific polymerase chain reaction (PCR) assays, targeting the major surface protein 4 (msp4), were employed for identification of A. ovis and A. phagocytophilum and selected products were confirmed via sequencing. A total of 230 small ruminants (54.37%) were found to be infected with A. ovis and/or A. phagocytophilum. The rates of infected animals for A. ovis and A. phagocytophilum were 50.83% (215/423) and 8.51% (36/423) respectively. Coinfection rate in small ruminants was determined as 4.96% (21/423). Sequence diversity rates of 0-0.94% for A. ovis and 0.41-2.49% for A. phagocytophilum have been observed. This is the first detection of A. ovis and A. phagocytophilum in sheep and goats in Thrace region of northwestern Turkey via polymerase chain reaction and sequence characterization. Further researches are needed to determine the vectors, vector-host interactions and genotypic variants that may affect the presence and distribution of Anaplasma species in the region

PDF

___

Dumler JS, Barbet AF, Bekker CPJ, Dasch GA, Palmer GH, Ray SC, Rikihisa Y, Rurangirwa FR: Reorganisation of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and 'HGE agent' as subjective synonyms of Ehrlichia phagocytophila. Int J Syst Evol Microbiol, 51, 2145-2165, 2001.
Rikihisa Y: The tribe Ehrlichieae and ehrlichial diseases. Clin Microbiol Rev, 4, 286-308, 1991.
Rymaszewska A, Grenda S: Bacteria of the genus Anaplasma - characteristics of Anaplasma and their vectors: A review. Veterinarni Medicina, 53 (11): 573-584, 2008.
Friedhoff KT: Tick-borne diseases of sheep and goats caused by Babesia, Theileria or Anaplasma spp. Parassitologia, 39, 99-109, 1997.
Tibbitts T, Goff W, Foreyt W, Stiller D: Susceptibility of two rocky mountain bighorn sheep to experimental infection with Anaplasma ovis. J Wildl Dis, 28 (1): 125-129, 1992. DOI: 10.7589/0090-3558-28.1.125
Kocan KM, de la Fuente J, Blouin EF, Garcia-Garcia JC: Anaplasma marginale (Rickettsiales: Anaplasmataceae): Recent advances in defining host-pathogen adaptations of a tick-borne Rickettsia. Parasitology, 129, 300, 2004. DOI: 10.1017/S0031182003004700
Uilenberg G: International collaborative research: Significance of tick- borne hemoparasitic diseases to world animal health. Vet Parasitol, 57, 19- , 1995. DOI: 10.1016/0304-4017(94)03107-8
Ioannou I, Sandalakis V, Kassinis N, Chochlakis D, Papadopoulos B, Loukaides F, Tselentis Y, Psaroulaki A: Tick-borne bacteria in mouflons and their ectoparasites in Cyprus. J Wildl Dis, 47, 300-306, 2011. DOI: 7589/0090-3558-47.2.300
Liu Z, Ma M, Wang Z, Wang J, Peng Y, Li Y, Guan G, Luo J, Yin H: Molecular survey and genetic identification of Anaplasma species in goats from central and southern China. Appl Environ Microbiol, 78, 464- , 2012. DOI: 10.1128/AEM.06848-11 de la Fuente J, Atkinson MW, Hogg JT, Miller DS, Naranjo V,
Almazan C, Anderson N, Kocan KM: Genetic characterization of Anaplasma ovis strains from bighorn sheep in Montana. J Wildl Dis, 42, 385, 2006. DOI: 10.7589/0090-3558-42.2.381
Papadopoulos B: Cattle and small ruminant piroplasmosis in Macedonia, Greece. Parassitologia, 41 (Suppl 1): 81-84, 1999.
Psaroulaki A, Chochlakis D, Sandalakis V, Vranakis I, Ioannou I, Tselentis Y: Phylogentic analysis of Anaplasma ovis strains isolated from sheep and goats using groEL and mps4 genes. Vet Microbiol, 138, 394- , 2009. DOI: 10.1016/j.vetmic.2009.04.018
Hosseini-Vasoukolaei N, Oshaghi MA, Shayan P, Vatandoost H, Babamahmoudi F, Yaghoobi-Ershadi MR, Telmadarraiy Z, Mohtarami F: Anaplasma infection in ticks, livestock and human in Ghaemshahr, Mazandaran province, Iran. J Arthropod Borne Dis, 8 (2): 204-211, 2014.
Carrade DD, Foley JE, Borjesson DL, Sykes JE: Canine granulocytic Anaplasmosis: A review. J Vet Intern Med, 23, 1129-1141, 2009. DOI: 1111/j.1939-1676.2009.0384.x
Stuen S, Bergstrom K, Palmer E: Reduced weight gain due to subclinical Anaplasma phagocytophilum (formerly Ehrlichia phagocytophila) infection. Exp Appl Acarol, 28, 209-215, 2002. DOI: 10.1023/A:1025350517733
Woldehiwet Z, Scott GR: Tick-borne (pasture) fever. In, Woldehiwet Z, Ristic M (Eds): Rickettsial and Chlamydial Diseases of Domestic Animals. 254, Pergamon Press, Oxford, 1993.
Giudice E, Giannetto C, Torina A, Gianesella M: Anaplasma phagocytophilum ıntragranulocytic morulae in aborting sheep: A herd case in Sicily. Transbound Emerg Dis,58, 263-267, 2011. DOI: 10.1111/j.1865- 2010.01190.x
Yu XJ, Walker DH: The Order Rickettsiales. In, Dworkin M, Falkow S, Rosenberg E, Schleifer KH, Stackebrandt E (Eds): The Prokaryotes. 493- , Springer, New York, 2006.
Christova IS, Dumler JS: Human granulocytic ehrlichiosis in Bulgaria. Am J Trop Med Hyg, 60 (1): 58-61, 1999.
Daniel SA, Manika K, Arvanitidou M, Diza E, Symeonidis N, Antoniadis A: Serologic evidence of human granulocytic ehrlichiosis, Greece. Emerg Infect Dis, 8 (6): 643-644, 2002.
Bashiribod H, Kazemi B, Eslami G, Bigdeli S, Bandehpour M, Rahbarian N, Ramezani Z: First molecular detection of Anaplasma phagocytophilum in Ixodes ricinus ticks in Iran. J Med Sci, 4, 282-286, 2004. DOI: 10.3923/jms.2004.282.286
Ozcan HC: Ankara ve civarında evcil hayvanlarda görülen Piroplasmose vakaları ve tedavileri üzerinde araştırmalar. Doçentlik Tezi. Ankara Üniv Vet Fak Yay, 143, 83, 1961.
Sayın F, Dinçer Ş, Karaer Z, Çakmak A, Yukarı BA, Eren H, Değer S, Nalbantoğlu S: Status of the tick-borne diseases in sheep and goats in Turkey. Parasitologia, 39 (2): 153-156, 1997.
Gokce HI, Genc O, Akca A, Vatansever Z, Unver A, Erdogan HM: Molecular and serological evidence of Anaplasma phagocytophilum infection of farm animals in the Black Sea region of Turkey. Acta Vet Hung, , 281-292, 2008. DOI: 10.1556/AVet.56.2008.3.2
Altay K, Dumanli N, Aktas M, Ozubek S: Survey of Anaplasma infections in small ruminants from East part of Turkey. Kafkas Unv Vet Fak Derg, 20, 1-4, 2014. DOI: 10.9775/kvfd.2013.9189
Aktas M, Vatansever Z, Altay K, Aydin MF, Dumanli N: Molecular evidence for Anaplasma phagocytophilum in Ixodes ricinus from Turkey. T Roy Soc Trop Med H, 104, 10-15, 2010. DOI: 10.1016/j.trstmh.2009.07.025
Sen E, Uchishima Y, Okamoto Y, Fukui T, Kadosaka T, Ohashi N, Masuzawa T: Molecular detection of Anaplasma phagocytophilum and Borrelia burgdorferi in Ixodes ricinus ticks from Istanbul metropolitan area and rural Trakya (Thrace) region of north-western Turkey. Ticks Tick Borne Dis, 2, 94-98, 2011. DOI: 10.1016/j.ttbdis.2011.03.004
Aktas M: A survey of ixodid tick species and molecular identification of tickborne pathogens. Vet Parasitol, 200, 276-283, 2014. DOI: 10.1016/j. vetpar.2013.12.008
Aktas M, Altay K, Dumanli N: Molecular detection and identification of Anaplasma and Ehrlichia species in cattle from Turkey. Ticks and Tick- borne Dis, 2, 62-66, 2011. DOI: 10.1016/j.ttbdis.2010.11.002
Aktas M, Ozubek S: Bovine anaplasmosis in Turkey: First laboratory confirmed clinical cases caused by Anaplasma phagocytophilum. Vet Microbiol, 178, 246-251, 2015. DOI: 10.1016/j.vetmic.2015.05.021
Düzlü Ö, İnci A, Yıldırım A, Önder Z, Çiloğlu A: Köpeklerde kene kaynaklı bazı protozoon ve rickettsial enfeksiyonların real time PCR ile araştırılması ve saptanan izolatların moleküler karakterizasyonları. Ankara Univ Vet Fak Derg, 61, 275-282, 2014. DOI: 10.1501/Vetfak_0000002642
Aktas M, Altay K, Dumanli N, Kalkan A: Molecular detection and identification of Erlichia and Anaplasma species in Ixodid ticks. Parasitol Res, 104, 1243-1248, 2009. DOI: 10.1007/s00436-009-1377-1
Dreher UM, de la Fuente J, Hofmann-Lehmann R, Meli ML, Pusterla N, Kocan KM, Woldehiwet Z, Braun U, Regula G, Staerk KD, Lutz H: Serologic cross-reactivity between Anaplasma marginale and Anaplasma phagocytophilum. Clin Diagn Lab Immunol, 12, 1177-1183, DOI: 10.1128/CDLI.12.10.1177-1183.2005
Ogden NH, Casey AN, Woldehiwet Z, French NP: Transmission of Anaplasma phagocytophilum to Ixodes ricinus ticks from sheep in the acute and post-acute phases of infection. Infect Immun, 71, 2071-2078, DOI: 10.1128/IAI.71.4.2071-2078.2003 de la Fuente VJ, Torina A, Caracappa S, Tumino G, Furla R, Almaza
C, Kocan KM: Serologic and molecular characterization of Anaplasma species infection in farm animals and ticks from Sicily. Vet Parasitol, 133, 362, 2005. DOI: 10.1016/j.vetpar.2005.05.063
Torina A, Agnone A, Blanda V, Alongi A, D'Agostino R, Caracappa S, Marino AM, Di Marco V, de la Fuente J: Development and validation of two PCR tests for the detection of and differentiation between Anaplasma ovis and Anaplasma marginale. Ticks Tick Borne Dis, 3, 283-287, DOI: 10.1016/j.ttbdis.2012.10.033 de la Fuente J, Massung RF, Wong SJ, Chu FK, Lutz H, Meli M, von
Loewenich FD, Grzeszczuk A, Torina A, Caracappa S, Mangold AJ, Naranjo V, Stuen S, Kocan KM: Sequence analysis of the msp4 gene of Anaplasma phagocytophilum strains. J Clin Microbiol, 43, 1309-1317, 2005. DOI: 10.1128/JCM.43.3.1309-1317.2005
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S: MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance and maximum parsimony methods. Mol Biol Evol, 28, 2731-2739, 2011. DOI: 10.1093/molbev/msr121
Woldehiwet Z: The natural history of Anaplasma phagocytophilum. Vet Parasitol, 167, 108-122, 2010. DOI: 10.1016/j.vetpar.2009.09.013
Chastagner A, Dugat T, Vourc'h G, Verheyden H, Legrand L, Bachy V, Chabanne L, Joncour G, Maillard R, Boulouis H-J, Haddad N, Bailly X, Leblond A: Multilocus sequence analysis of Anaplasma phagocytophilum reveals three distinct lineages with different host ranges in clinically ill French cattle. Vet Res, 45, 114, 2014. DOI: 10.1186/s13567-014-0114-7
Strasek Smrdel K, von Loewenich FD, Petrovec M, Avsic Zupanc T: Diversity of ankA and msp4 genes of Anaplasma phagocytophilum in Slovenia. Ticks Tick Borne Dis, 6, 164-166, 2015. DOI: 10.1016/j.ttbdis. 11.008
Aydın L, Bakirci S: Geographical distribution of ticks in Turkey. Parasitol Res, 101 (Suppl. 2): 163-166, 2007. DOI: 10.1007/s00436-007- 5
Vatansever Z, Gargili A, Aysul NS, Sengoz Z, Estrada-Peña A: Ticks biting humans in the urban area of Istanbul. Parasitol Res, 102, 551-553, DOI: 10.1007/s00436-007-0809-z
Gargili A, Kar S, Yilmazer N, Cerit C, Sonmez G, Sahin F, Gunseli Alp H, Vatansever Z: Evaluation of ticks biting humans in Thrace province, Turkey. Kafkas Univ Vet Fak Derg, 16 (Suppl-A): S141-S146, 2010. DOI: 10.9775/kvfd.2009.2555