KAYSERİ YÖRESİNDEKİ IXODID KENE POPÜLASYONLARINDA ANAPLASMA PHAGOCYTOPHILUM’UN MOLEKÜLER KARAKTERİZASYONU
Öz
Bu çalışma, Kayseri yöresindeki sığırlardan toplanmış ixodid kenelerde Anaplasma phagocytophilum’un Real Time PCR’la araştırılması ve pozitif izolatların 16S rRNA gen bölgesi yönünden karakterize edilmesi amacıyla yapılmıştır. Bu amaçla sığırlardan toplanmış 265 ergin ixodid keneden genomik DNA ekstrakte edilmiştir. DNA ekstraksiyonu sonrası 16S rRNA gen bölgesinin 641 bp’lik kısmını amplifiye eden Anaplasma phagocytophilum spesifik primerlerle nested PCR analizleri yapılmıştır. Real Time PCR’la bireysel olarak incelemesi yapılan örneklerden bir H. marginatum ve bir R. turanicus örneğinde pozitiflik saptanmıştır. DNA dizilerinin GenBank kayıtları gerçekleştirilmiş ve mevcut diğer bazı A. phagocytophilum suşları ile hizalamaları yapılarak filogenisi araştırılmıştır. Filogenetik analiz sonucu Kayseri yöresindeki sığırlardan toplanmış H. marginatum ve R. turanicus örneklerinde belirlenen A. phagocytophilum izolatları ile Dünya’dan ve Türkiye’den daha önce GenBank’a girilmiş izolatların 3 ana dalda (A, B ve C kümeleri) kümelendikleri belirlenmiştir. A ve B gruplarındaki A. phagocytophilum izolatlarının homolog oldukları, C grubunda yer alan izolatların ise kendi aralarında ortalama %0,2’lik genetik farklılığın bulunduğu belirlenmiştir. A grubundaki izolatların B grubundakilerle %10,8±2,0, C grubundakilerle ise %13,0±2,7; B grubundaki izolatların C grubundaki izolatlarla arasındaki ortalama genetik farklılık %13,8±2,8 saptanmıştır. Sonuç olarak bu çalışma ile Kayseri yöresindeki sığırlardan toplanmış kene örneklerinde A. phagocytophilum’un moleküler yaygınlığı ve genetik karakterleri hakkında bilimsel veriler elde edilmiştir.
Anahtar Kelimeler
Anaplasma phagocytophilum Kayseri moleküler karakterizasyon kene 16S rRNA
Destekleyen Kurum
Erciyes Üniversitesi Bilimsel Araştırma Projeleri Koordinatörlüğü
Proje Numarası
TYL-2014-5506
Kaynakça
- Stephenson N, Hodzic E, Mapes S, Rejmanek D, Foley J. A real-time PCR assay for differentiating pathogenic Anaplasma phagocytophilum from an apathogenic, woodrat-adapted genospecies from North America. Ticks Tick Borne Dis 2015; 6(6):774-778.
- Aktas M. A survey of ixodid tick species and molecular identification of tick-borne pathogens. Vet Parasitol 2014; 200(3-4):276-783.
- Scharf W, Schauer S, Freyburger F, et al. Distinct host species correlate with Anaplasma phagocytophilum ankA gene clusters. J Clin Microbiol 2011; 49(3):790-796.
- Mwale R, Mulavu M, Khumalo CS, et al. Molecular detection and characterization of Anaplasma spp. in cattle and sable antelope from Lusaka and North-Western provinces of Zambia. Vet Parasitol Reg Stud Reports 2023; 39:100847.
- Van Loo H, Pascottini OB, Hooyberghs J, et al. Detection of Anaplasma phagocytophilum in fetal and placental tissue of bovine abortions and perinatal mortalities. Vet Rec 2023; e2880.
- Estrada-Pena A, Bouattour A, Camicas JL, Walker AR. Ticks of domestic animals in the Mediterranean Region (1th ed). University of Zaragoza, Spain 2004.
- Courtney JW, Kostelnik LM, Zeidner NS, Massung RF. Multiplex real-time PCR for detection of Anaplasma phagocytophilum and Borrelia burgdorferi. J Clin Microbiol 2004; 42(7):3164-3168.
- Kawahara M, Rikihisa Y, Lin Q, et al. Novel genetic variants of Anaplasma phagocytophilum, Anaplasma bovis, Anaplasma centrale, and a novel Ehrlichia sp. in wild deer and ticks on two major islands in Japan. Appl Environ Microbiol 2006; 72(2):1102-1109.
- Drummond AJ, Ashton B, Buxton S. Geneious v5.5. http://www.geneious.com. 2011.
- Tamura K, Stecher G, Peterson D, Filipski A., Kumar S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 2013; 30:2725-2729.
- Dantas-Torres F, Chomel BB, Otranto D. Ticks and tick-borne diseases: A one health perspective. Trends Parasitol 2012; 28:437-446.
- Arslan ÖM. Türkiye’de hayvanlarda kene enfestasyonları ve kenelerin bulaştırdığı hastalıkların durumu, 14. Ulusal Parazitoloji Kongresi Bildiri Kitabı, İzmir 18-25 Eylül 2005; YM02-04.
- Aydın L, Bakırcı S. Geographical distribution of ticks in Turkey. Parasitol Res 2007; 101:163-166.
- Dumanlı N, Altay K, Aydın MF. Türkiye’de sığır, koyun ve keçilerde belirlenen kene türleri. Türkiye Klinikleri Vet Bil Derg 2012; 3(2): 67-72.
- Karaer Z, Yukarı BA, Aydın L. Türkiye keneleri ve vektörlükleri. In: Özcel MA, Daldal N (eds), Parazitolojide Artropod Hastalıkları ve Vektörler. Türkiye Parazitoloji Derneği 1997; ss 363-433.
- Stuen S. Tick-borne infections in small ruminants in northern Europe. Small Rum Res 2013; 110:142-144.
- Coskun A, Ekici OD, Guzelbektes H, Aydogdu U, Sen I. Acute phase proteins, clinical, hematological and biochemical parameters in dairy cows naturally ınfected with Anaplasma marginale. Kafkas Univ Vet Fak Derg 2012; 18(3):497-502.
- Aktas M, Altay K, Dumanli N. Molecular detection and identification of Anaplasma and Ehrlichia species in cattle from Turkey. Ticks Tick Borne Dis 2011; 2(1):62-65.
- Aktas M, Altay K, Ozubek S, Dumanli N. A survey of ixodid ticks feeding on cattle and prevalence of tick-borne pathogens in the Black Sea region of Turkey. Vet Parasitol 2012; 187(3-4):567-571.
- Sen E, Uchishima Y, Okamoto Y, et al. 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 2011; 2(2):94-98.
- Hoşgör M, Bilgiç HB, Bakırcı S, Ünlü AH, Karagenç T, Eren H. Detection of Anaplasma/Ehrlichia species of cattle and ticks in Aydin region. Turkiye Parazitol Derg 2015; 39(4):291-298.
- Noaman V, Shayan P. A new PCR-RFLP method for detection of Anaplasma marginale based on 16S rRNA. Vet Res Commun 2010; 34: 43-50.
- Noaman V, Shayan P. Molecular detection of Anaplasma bovis in cattle from central part of Iran. Vet Res 2010; 1(2):117-122.
- Chen SM, Dumler JS, Bakken JS, Walker DH. Identification of a granulocytoyropic Ehrlichia species as the etiologic agent of human disease. J Clin Microbiol 1994; 32:589-595.
- Bakken JS, Haller I, Riddell D, Walls JJ, Dumler JS. The serological response of patients with the agent of human granulocytic ehrlichiosis. Clin Infect Dis 2002; 34:22-27.
- Comer JS, Nicholson WL, Sumner JW, Olson JG, Childs JE. Diagnosis of human ehrlichiosis by PCR assay of acute-phase serum. J Clin Microbiol 1999; 37:31-34.
- Petrovec M, Lotric Furlan S, et al. Human disease in Europe caused by a granulocytic Ehrlichia species. J Clin Microbiol 1997; 35:1556-1559.
- Goodman JL, Nelson C, Vitale B, et al. Direct cultivation of the causative agent of human granulocytic ehrlichiosis. N Engl J Med 1996; 334:210-215.
Öz
The study was conducted to investigate the presence of Anaplasma phagocytophilum in ixodid ticks collected from cattle in Kayseri region of Turkey using Real Time PCR and to characterize positive isolates based on 16S rRNA gene region. DNA was extracted from 265 adult ticks. Nested PCR analyses were performed using Anaplasma phagocytophilum-specific primers amplifying a 641 bp fragment of 16S rRNA gene region. Real Time PCR analysis revealed positive results in one H. marginatum and R. turanicus sample. DNA sequences were submitted to GenBank and analyzed by pairwise and multiple sequence alignments with other A. phagocytophilum strains in GenBank to investigate the phylogeny. The phylogenetic analysis revealed that A. phagocytophilum isolates collected from H. marginatum and R. turanicus samples in Kayseri region clustered into three main groups (A, B, and C) with previously reported isolates from the world. A and B groups showed high homology, whereas C group had an average genetic variation of 0.2%. The average genetic differences between A and B groups were 10.8±2.0 and 13.0±2.7% between A and C groups, while the average genetic difference between B and C groups was 13.8±2.8%. In conclusion, this study provides scientific data on molecular prevalence and genetic characteristics of A. phagocytophilum in tick samples in Turkey.
Anahtar Kelimeler
Anaplasma phagocytophilum Kayseri molecular characterization tick 16S rRNA
Proje Numarası
TYL-2014-5506
Kaynakça
- Stephenson N, Hodzic E, Mapes S, Rejmanek D, Foley J. A real-time PCR assay for differentiating pathogenic Anaplasma phagocytophilum from an apathogenic, woodrat-adapted genospecies from North America. Ticks Tick Borne Dis 2015; 6(6):774-778.
- Aktas M. A survey of ixodid tick species and molecular identification of tick-borne pathogens. Vet Parasitol 2014; 200(3-4):276-783.
- Scharf W, Schauer S, Freyburger F, et al. Distinct host species correlate with Anaplasma phagocytophilum ankA gene clusters. J Clin Microbiol 2011; 49(3):790-796.
- Mwale R, Mulavu M, Khumalo CS, et al. Molecular detection and characterization of Anaplasma spp. in cattle and sable antelope from Lusaka and North-Western provinces of Zambia. Vet Parasitol Reg Stud Reports 2023; 39:100847.
- Van Loo H, Pascottini OB, Hooyberghs J, et al. Detection of Anaplasma phagocytophilum in fetal and placental tissue of bovine abortions and perinatal mortalities. Vet Rec 2023; e2880.
- Estrada-Pena A, Bouattour A, Camicas JL, Walker AR. Ticks of domestic animals in the Mediterranean Region (1th ed). University of Zaragoza, Spain 2004.
- Courtney JW, Kostelnik LM, Zeidner NS, Massung RF. Multiplex real-time PCR for detection of Anaplasma phagocytophilum and Borrelia burgdorferi. J Clin Microbiol 2004; 42(7):3164-3168.
- Kawahara M, Rikihisa Y, Lin Q, et al. Novel genetic variants of Anaplasma phagocytophilum, Anaplasma bovis, Anaplasma centrale, and a novel Ehrlichia sp. in wild deer and ticks on two major islands in Japan. Appl Environ Microbiol 2006; 72(2):1102-1109.
- Drummond AJ, Ashton B, Buxton S. Geneious v5.5. http://www.geneious.com. 2011.
- Tamura K, Stecher G, Peterson D, Filipski A., Kumar S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 2013; 30:2725-2729.
- Dantas-Torres F, Chomel BB, Otranto D. Ticks and tick-borne diseases: A one health perspective. Trends Parasitol 2012; 28:437-446.
- Arslan ÖM. Türkiye’de hayvanlarda kene enfestasyonları ve kenelerin bulaştırdığı hastalıkların durumu, 14. Ulusal Parazitoloji Kongresi Bildiri Kitabı, İzmir 18-25 Eylül 2005; YM02-04.
- Aydın L, Bakırcı S. Geographical distribution of ticks in Turkey. Parasitol Res 2007; 101:163-166.
- Dumanlı N, Altay K, Aydın MF. Türkiye’de sığır, koyun ve keçilerde belirlenen kene türleri. Türkiye Klinikleri Vet Bil Derg 2012; 3(2): 67-72.
- Karaer Z, Yukarı BA, Aydın L. Türkiye keneleri ve vektörlükleri. In: Özcel MA, Daldal N (eds), Parazitolojide Artropod Hastalıkları ve Vektörler. Türkiye Parazitoloji Derneği 1997; ss 363-433.
- Stuen S. Tick-borne infections in small ruminants in northern Europe. Small Rum Res 2013; 110:142-144.
- Coskun A, Ekici OD, Guzelbektes H, Aydogdu U, Sen I. Acute phase proteins, clinical, hematological and biochemical parameters in dairy cows naturally ınfected with Anaplasma marginale. Kafkas Univ Vet Fak Derg 2012; 18(3):497-502.
- Aktas M, Altay K, Dumanli N. Molecular detection and identification of Anaplasma and Ehrlichia species in cattle from Turkey. Ticks Tick Borne Dis 2011; 2(1):62-65.
- Aktas M, Altay K, Ozubek S, Dumanli N. A survey of ixodid ticks feeding on cattle and prevalence of tick-borne pathogens in the Black Sea region of Turkey. Vet Parasitol 2012; 187(3-4):567-571.
- Sen E, Uchishima Y, Okamoto Y, et al. 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 2011; 2(2):94-98.
- Hoşgör M, Bilgiç HB, Bakırcı S, Ünlü AH, Karagenç T, Eren H. Detection of Anaplasma/Ehrlichia species of cattle and ticks in Aydin region. Turkiye Parazitol Derg 2015; 39(4):291-298.
- Noaman V, Shayan P. A new PCR-RFLP method for detection of Anaplasma marginale based on 16S rRNA. Vet Res Commun 2010; 34: 43-50.
- Noaman V, Shayan P. Molecular detection of Anaplasma bovis in cattle from central part of Iran. Vet Res 2010; 1(2):117-122.
- Chen SM, Dumler JS, Bakken JS, Walker DH. Identification of a granulocytoyropic Ehrlichia species as the etiologic agent of human disease. J Clin Microbiol 1994; 32:589-595.
- Bakken JS, Haller I, Riddell D, Walls JJ, Dumler JS. The serological response of patients with the agent of human granulocytic ehrlichiosis. Clin Infect Dis 2002; 34:22-27.
- Comer JS, Nicholson WL, Sumner JW, Olson JG, Childs JE. Diagnosis of human ehrlichiosis by PCR assay of acute-phase serum. J Clin Microbiol 1999; 37:31-34.
- Petrovec M, Lotric Furlan S, et al. Human disease in Europe caused by a granulocytic Ehrlichia species. J Clin Microbiol 1997; 35:1556-1559.
- Goodman JL, Nelson C, Vitale B, et al. Direct cultivation of the causative agent of human granulocytic ehrlichiosis. N Engl J Med 1996; 334:210-215.
Ayrıntılar
| Birincil Dil | İngilizce |
|---|---|
| Konular | Veteriner Parazitoloji |
| Bölüm | Araştırma Makalesi |
| Yazarlar | |
| Proje Numarası | TYL-2014-5506 |
| Erken Görünüm Tarihi | 3 Kasım 2023 |
| Yayımlanma Tarihi | 4 Aralık 2023 |
| Gönderilme Tarihi | 6 Nisan 2023 |
| Yayımlandığı Sayı | Yıl 2023 Cilt: 32 Sayı: 3 |
