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First Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Muş Province of Turkey

Year 2021, Volume: 7 Issue: 3, 312 - 318, 01.11.2021
https://doi.org/10.30934/kusbed.895438

Abstract

Objective: Equine granulocytic anaplasmosis (EGA) caused by Anaplasma phagocytophilum is a disease that generally develop characteristic high fever and a number of changes in blood such as thrombocytopenia. The main transmission of A. phagocytophilum is hard ticks through intrastadially. The aim of this study was to determine the presence of A. phagocytophilum in horses in Muş province of Turkey by using molecular methods.
Methods: In the present survey, blood samples were collected in Muş province from 93 horse. Commercial ELISA kit (IgG) and nested-PCR were performed. The DNA was extracted from blood and analysed by nested-PCR technique targeting 16S rRNA gene of A. phagocytophilum and then positive PCR products were sequenced.
Results: The seroprevalence against A. phagocytophilum was 8.6% and 6 horses (6.4%) showed positive nested-PCR results. The sequences had 99.5 and 100% homology with the BLAST published A. phagocytophilum amplicons.
Conclusion: To the best of our knowledge, this is the first molecular data of A. phagocytophilum in horses in Muş province of Turkey. This study also reveals that A. phagocytophilum isolates isolated from a small region are closely related to A. phagocytophilum isolated from a wide variety of hosts.

Supporting Institution

Van Yuzuncu Yil University Scientific Research Project Fund

Project Number

THD-2020-9287.

References

  • Inci A, Yildirim A, Duzlu O, Doganay M, Aksoy S. Tick-Borne Diseases in Turkey: A Review Based on One Health Perspective. PLoS Negl Trop Dis. 2016; 10(12):e0005021. doi: 10.1371/journal.pntd.0005021.
  • Oğuz B, Özdal N, Kılınç ÖO, Karakuş A, Çelik BA, Değer MS. Van, Muş, Siirt ve Diyarbakır İllerinde Sığırlarda Anaplasmosis’in Seroprevalansı. Kocatepe Vet J. 2018;11(3):208-214. doi.org/10.30607/kvj.397726.
  • Pusterla N, Madigan JE. Equine Granulocytic Anaplasmosis. J Equine Vet Sci. 2013;33:493-496. doi.org/10.1016/j.jevs.2013.03.188.
  • Ceylan O, Byamukama B, Ceylan C, et al. Tick-Borne Hemoparasites of Sheep: A Molecular Research in Turkey. Pathogens. 2021;10(2):162. doi: 10.3390/pathogens10020162.
  • Fuente JDL, Massung RF, Wong SJ, et al. Sequence Analysis of the msp4 Gene of Anaplasma phagocytophilum Strains. J Clin Microbiol. 2005;43(3):1309-1317. doi: 10.1128/JCM.43.3.1309-1317.2005.
  • Wang Y, Chen C, Zhang L. Molecular characterization of Msp2/P44 of Anaplasma phagocytophilum isolated from infected patients and Haemaphysalis longicornis in Laizhou Bay, Shandong Province, China. PLoS One. 2013;8:e78189. doi: 10.1371/journal.pone.0078189.
  • Smrdel KS, von Loewenich FD, Petrovec M, Županc TA. Diversity of ankA and msp4 genes of Anaplasma phagocytophilum in Slovenia. Ticks Tick Borne Dis. 2015;6(2):164-166. doi.org/10.1016/j.ttbdis.2014.11.008.
  • Chisu V, Zobba R, Lecis R, et al. GroEL typing and phylogeny of Anaplasma species in ticks from domestic and wild vertebrates. Ticks Tick Borne Dis. 2018; 9:31-36. doi: 10.1016/j.ttbdis.2017.10.012.
  • Akkoyun Z, Oğuz B. Seroprevalence of Theileria equi and Babesia caballi in horses of Mus province, Turkey. AJVS. 2019;60:22-29. doi: 10.5455/ajvs.22338.
  • Machado RZ, Teixeira MM, Rodrigues AC, et al. Molecular diagnosis and genetic diversity of tick-borne Anaplasmataceae agents infecting the African buffalo Syncerus caffer from Marromeu Reserve in Mozambique. Parasit Vectors. 2016;9:454. doi: 10.1186/s13071-016-1715-y.
  • Dzięgiel B, Adaszek L, Carbonero A, et al. Comparative analysis of 16S RNA nucleotide sequences of Anaplasma phagocytophilum detected in the blood of horses from various parts of Europe. J Med Microbiol. 2013;62:1891-1896. doi: 10.1099/jmm.0.058636-0.
  • Yang J, Liu Z, Niu Q, et al. Evaluation of different nested PCRs for detection of Anaplasma phagocytophilum in ruminants and ticks. BMC Vet Res. 2016;12(1):35. doi: 10.1186/s12917-016-0663-2.
  • Günaydın E, Pekkaya S, Kuzugüden F, Zeybek M, Gökmen TG, Ütük AE. The First Detection of anti-Anaplasma phagocytophilum Antibodies in Horses in Turkey. Kafkas Univ Vet Fak Derg. 2018;24(6):867-871. doi: 10.9775/kvfd.2018.20171.
  • 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. doi: 10.1128/AEM.72.2.1102-1109.2006.
  • Hall T. Bio Edit: a user-friendly biological sequence alignment editor and analysis suite. Nucleic Acids Symp Ser. 1999;41:95-98. doi: 10.14601/Phytopathol_Mediterr-14998u1.29.
  • Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol. 2016;33(7):1870-1874. doi: 10.1093/molbev/msw054.
  • M’ghirbi Y, Yaïch H, Ghorbel A, Bouattour A. Anaplasma phagocytophilum in horses and ticks in Tunisia. Parasit Vectors. 2012;5:180. doi: 10.1186/1756-3305-5-180.
  • Laamari A, Azzag N, Tennah S, et al. Seroprevalence of Antibodies Against Anaplasma Phagocytophilum and Borrelia Burgdorferi in Horses (Equus Caballus) from Northern Algeria. J Vet Res. 2020;64(3):413-419. doi: 10.2478/jvetres-2020-0045.
  • Praskova I, Bezdekova B, Zeman P, Jahn P. Seroprevalence of Anaplasma phagocytophilum in horses in the Czech Republic. Ticks Tick Borne Dis. 2011;2:111-115. doi.org/10.1016/j.ttbdis.2011.01.002.
  • Lee SH, Kim KT, Yun SH, et al. Serological and molecular detection of Anaplasma phagocytophilum in horse reared in Korea. Vet Med. 2015;60:533-538. doi: 10.17221/8491-VETMED.
  • Laus F, Veronesi F, Passamonti F, et al. Prevalence of tick borne pathogens in horses from Italy. J Vet Med Sci. 2013;75:715-720. doi: 10.1292/jvms.12-0449.
  • Andel AEV, Magnarelli LA, Heimer R, Wilson ML. Development and duration of antibody response against Ehrlichia equi in horses. J Am Vet Med Assoc. 1998;212:1910-1914. PMID: 9638192
  • Passamonti F, Veronesi F, Cappelli K, et al. Anaplasma phagocytophilum in horses and ticks: a preliminary survey of Central Italy. Comp Immunol Microbiol Infect Dis. 2010;33:73-83. doi: 10.1016/j.cimid.2008.08.002.
  • Altay K, Dumanlı N, Aktas M, Özübek S. Survey of Anaplasma infections in small ruminants from East part of Turkey. Kafkas Univ Vet Fak Derg. 2014;20:1-4. doi: 10.9775/kvfd.2013.9189.
  • Seo MG, Ouh IO, Choi E, Kwon OD, Kwak D. Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Korea. Korean J Parasitol. 2018;56(6):559-565. doi: 10.3347/kjp.2018.56.6.559.
  • Haschke-Becher E, Bernauer R, Walleczek AM, et al. First detection of the Anaplasma phagocytophilum groEL-A genotype in man. J Infect. 2010;60(4):300-305. doi: 10.1016/j.jinf.2009.12.010.
  • Rymaszewska A. Genotyping of Anaplasma phagocytophilum strains from Poland for selected genes. Folia Biol. 2014;62:37-48. doi: 10.3409/fb62_1.37.
  • Domenico MD, Pascucci I, Curini V, et al. Detection of Anaplasma phagocytophilum genotypes that are potentially virulent for human in wild ruminants and Ixodes ricinus in Central Italy. Ticks Tick Borne Dis. 2016;7:782-787. doi: 10.1016/j.ttbdis.2016.03.012.
  • Kolo AO, Collins NE, Brayton KA, et al. Anaplasma phagocytophilum and Other Anaplasma spp. in Various Hosts in the Mnisi Community, Mpumalanga Province, South Africa. Microorganisms. 2020;8(11):1812. doi.org/10.3390/microorganisms8111812.

Türkiye’nin Muş İlinde Atlarda Anaplasma phagocytophilum’un İlk Moleküler Tespiti ve Filogenetik Analizi

Year 2021, Volume: 7 Issue: 3, 312 - 318, 01.11.2021
https://doi.org/10.30934/kusbed.895438

Abstract

Amaç: Anaplasma phagocytophilum'un neden olduğu equine granülositik anaplasmosis (EGA), genellikle karakteristik yüksek ateş ve trombositopeni gibi kanda bir dizi değişiklik geliştiren bir hastalıktır. A. phagocytophilum' un biyolojik bulaşması intrastadial yolla sert keneler tarafından yapılmaktadır. Bu çalışmanın amacı, Türkiye'nin Muş ilinde atlarda A. phagocytophilum varlığının moleküler yöntemlerle belirlenmesidir.
Yöntem: Bu çalışma için Muş ilinde bakılan 93 attan kan örnekleri alındı. Ticari ELISA kiti (IgG) ve nested PCR yapıldı. DNA’lar kandan ekstrakte edildi ve A. phagocytophilum'un 16S rRNA genini hedef alan nested PCR tekniği ile analiz edildi. Daha sonra pozitif PCR ürünleri sekanslandı.
Bulgular: A. phagocytophilum'a karşı seroprevalans % 8,6 bulunurken, 6 atın (% 6,4) nested PCR sonuçları pozitif bant gösterdi. Elde edilen sekanslar, BLAST tarafından yayınlanan A. phagocytophilum amplikonlar ile %99,5 ve %100 benzerlik gösterdi.
Sonuç: Yaptığımız literatür taramalarına göre bu çalışma, Türkiye'nin Muş ilinde atlarda A. phagocytophilum'un ilk moleküler verisidir. Aynı zamanda, küçük bir bölgeden izole edilen A. phagocytophilum izolatlarının, çok çeşitli konakçılardan izole edilen A. phagocytophilum ile yakından ilişkili olduğunu ortaya koymaktadır.

Project Number

THD-2020-9287.

References

  • Inci A, Yildirim A, Duzlu O, Doganay M, Aksoy S. Tick-Borne Diseases in Turkey: A Review Based on One Health Perspective. PLoS Negl Trop Dis. 2016; 10(12):e0005021. doi: 10.1371/journal.pntd.0005021.
  • Oğuz B, Özdal N, Kılınç ÖO, Karakuş A, Çelik BA, Değer MS. Van, Muş, Siirt ve Diyarbakır İllerinde Sığırlarda Anaplasmosis’in Seroprevalansı. Kocatepe Vet J. 2018;11(3):208-214. doi.org/10.30607/kvj.397726.
  • Pusterla N, Madigan JE. Equine Granulocytic Anaplasmosis. J Equine Vet Sci. 2013;33:493-496. doi.org/10.1016/j.jevs.2013.03.188.
  • Ceylan O, Byamukama B, Ceylan C, et al. Tick-Borne Hemoparasites of Sheep: A Molecular Research in Turkey. Pathogens. 2021;10(2):162. doi: 10.3390/pathogens10020162.
  • Fuente JDL, Massung RF, Wong SJ, et al. Sequence Analysis of the msp4 Gene of Anaplasma phagocytophilum Strains. J Clin Microbiol. 2005;43(3):1309-1317. doi: 10.1128/JCM.43.3.1309-1317.2005.
  • Wang Y, Chen C, Zhang L. Molecular characterization of Msp2/P44 of Anaplasma phagocytophilum isolated from infected patients and Haemaphysalis longicornis in Laizhou Bay, Shandong Province, China. PLoS One. 2013;8:e78189. doi: 10.1371/journal.pone.0078189.
  • Smrdel KS, von Loewenich FD, Petrovec M, Županc TA. Diversity of ankA and msp4 genes of Anaplasma phagocytophilum in Slovenia. Ticks Tick Borne Dis. 2015;6(2):164-166. doi.org/10.1016/j.ttbdis.2014.11.008.
  • Chisu V, Zobba R, Lecis R, et al. GroEL typing and phylogeny of Anaplasma species in ticks from domestic and wild vertebrates. Ticks Tick Borne Dis. 2018; 9:31-36. doi: 10.1016/j.ttbdis.2017.10.012.
  • Akkoyun Z, Oğuz B. Seroprevalence of Theileria equi and Babesia caballi in horses of Mus province, Turkey. AJVS. 2019;60:22-29. doi: 10.5455/ajvs.22338.
  • Machado RZ, Teixeira MM, Rodrigues AC, et al. Molecular diagnosis and genetic diversity of tick-borne Anaplasmataceae agents infecting the African buffalo Syncerus caffer from Marromeu Reserve in Mozambique. Parasit Vectors. 2016;9:454. doi: 10.1186/s13071-016-1715-y.
  • Dzięgiel B, Adaszek L, Carbonero A, et al. Comparative analysis of 16S RNA nucleotide sequences of Anaplasma phagocytophilum detected in the blood of horses from various parts of Europe. J Med Microbiol. 2013;62:1891-1896. doi: 10.1099/jmm.0.058636-0.
  • Yang J, Liu Z, Niu Q, et al. Evaluation of different nested PCRs for detection of Anaplasma phagocytophilum in ruminants and ticks. BMC Vet Res. 2016;12(1):35. doi: 10.1186/s12917-016-0663-2.
  • Günaydın E, Pekkaya S, Kuzugüden F, Zeybek M, Gökmen TG, Ütük AE. The First Detection of anti-Anaplasma phagocytophilum Antibodies in Horses in Turkey. Kafkas Univ Vet Fak Derg. 2018;24(6):867-871. doi: 10.9775/kvfd.2018.20171.
  • 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. doi: 10.1128/AEM.72.2.1102-1109.2006.
  • Hall T. Bio Edit: a user-friendly biological sequence alignment editor and analysis suite. Nucleic Acids Symp Ser. 1999;41:95-98. doi: 10.14601/Phytopathol_Mediterr-14998u1.29.
  • Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol. 2016;33(7):1870-1874. doi: 10.1093/molbev/msw054.
  • M’ghirbi Y, Yaïch H, Ghorbel A, Bouattour A. Anaplasma phagocytophilum in horses and ticks in Tunisia. Parasit Vectors. 2012;5:180. doi: 10.1186/1756-3305-5-180.
  • Laamari A, Azzag N, Tennah S, et al. Seroprevalence of Antibodies Against Anaplasma Phagocytophilum and Borrelia Burgdorferi in Horses (Equus Caballus) from Northern Algeria. J Vet Res. 2020;64(3):413-419. doi: 10.2478/jvetres-2020-0045.
  • Praskova I, Bezdekova B, Zeman P, Jahn P. Seroprevalence of Anaplasma phagocytophilum in horses in the Czech Republic. Ticks Tick Borne Dis. 2011;2:111-115. doi.org/10.1016/j.ttbdis.2011.01.002.
  • Lee SH, Kim KT, Yun SH, et al. Serological and molecular detection of Anaplasma phagocytophilum in horse reared in Korea. Vet Med. 2015;60:533-538. doi: 10.17221/8491-VETMED.
  • Laus F, Veronesi F, Passamonti F, et al. Prevalence of tick borne pathogens in horses from Italy. J Vet Med Sci. 2013;75:715-720. doi: 10.1292/jvms.12-0449.
  • Andel AEV, Magnarelli LA, Heimer R, Wilson ML. Development and duration of antibody response against Ehrlichia equi in horses. J Am Vet Med Assoc. 1998;212:1910-1914. PMID: 9638192
  • Passamonti F, Veronesi F, Cappelli K, et al. Anaplasma phagocytophilum in horses and ticks: a preliminary survey of Central Italy. Comp Immunol Microbiol Infect Dis. 2010;33:73-83. doi: 10.1016/j.cimid.2008.08.002.
  • Altay K, Dumanlı N, Aktas M, Özübek S. Survey of Anaplasma infections in small ruminants from East part of Turkey. Kafkas Univ Vet Fak Derg. 2014;20:1-4. doi: 10.9775/kvfd.2013.9189.
  • Seo MG, Ouh IO, Choi E, Kwon OD, Kwak D. Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Korea. Korean J Parasitol. 2018;56(6):559-565. doi: 10.3347/kjp.2018.56.6.559.
  • Haschke-Becher E, Bernauer R, Walleczek AM, et al. First detection of the Anaplasma phagocytophilum groEL-A genotype in man. J Infect. 2010;60(4):300-305. doi: 10.1016/j.jinf.2009.12.010.
  • Rymaszewska A. Genotyping of Anaplasma phagocytophilum strains from Poland for selected genes. Folia Biol. 2014;62:37-48. doi: 10.3409/fb62_1.37.
  • Domenico MD, Pascucci I, Curini V, et al. Detection of Anaplasma phagocytophilum genotypes that are potentially virulent for human in wild ruminants and Ixodes ricinus in Central Italy. Ticks Tick Borne Dis. 2016;7:782-787. doi: 10.1016/j.ttbdis.2016.03.012.
  • Kolo AO, Collins NE, Brayton KA, et al. Anaplasma phagocytophilum and Other Anaplasma spp. in Various Hosts in the Mnisi Community, Mpumalanga Province, South Africa. Microorganisms. 2020;8(11):1812. doi.org/10.3390/microorganisms8111812.
There are 29 citations in total.

Details

Primary Language English
Subjects Veterinary Surgery
Journal Section Original Article / Veterinary Sciences
Authors

Bekir Oğuz 0000-0001-8720-3940

Project Number THD-2020-9287.
Publication Date November 1, 2021
Submission Date March 11, 2021
Acceptance Date August 9, 2021
Published in Issue Year 2021 Volume: 7 Issue: 3

Cite

APA Oğuz, B. (2021). First Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Muş Province of Turkey. Kocaeli Üniversitesi Sağlık Bilimleri Dergisi, 7(3), 312-318. https://doi.org/10.30934/kusbed.895438
AMA Oğuz B. First Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Muş Province of Turkey. KOU Sag Bil Derg. November 2021;7(3):312-318. doi:10.30934/kusbed.895438
Chicago Oğuz, Bekir. “First Molecular Detection and Phylogenetic Analysis of Anaplasma Phagocytophilum in Horses in Muş Province of Turkey”. Kocaeli Üniversitesi Sağlık Bilimleri Dergisi 7, no. 3 (November 2021): 312-18. https://doi.org/10.30934/kusbed.895438.
EndNote Oğuz B (November 1, 2021) First Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Muş Province of Turkey. Kocaeli Üniversitesi Sağlık Bilimleri Dergisi 7 3 312–318.
IEEE B. Oğuz, “First Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Muş Province of Turkey”, KOU Sag Bil Derg, vol. 7, no. 3, pp. 312–318, 2021, doi: 10.30934/kusbed.895438.
ISNAD Oğuz, Bekir. “First Molecular Detection and Phylogenetic Analysis of Anaplasma Phagocytophilum in Horses in Muş Province of Turkey”. Kocaeli Üniversitesi Sağlık Bilimleri Dergisi 7/3 (November 2021), 312-318. https://doi.org/10.30934/kusbed.895438.
JAMA Oğuz B. First Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Muş Province of Turkey. KOU Sag Bil Derg. 2021;7:312–318.
MLA Oğuz, Bekir. “First Molecular Detection and Phylogenetic Analysis of Anaplasma Phagocytophilum in Horses in Muş Province of Turkey”. Kocaeli Üniversitesi Sağlık Bilimleri Dergisi, vol. 7, no. 3, 2021, pp. 312-8, doi:10.30934/kusbed.895438.
Vancouver Oğuz B. First Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Muş Province of Turkey. KOU Sag Bil Derg. 2021;7(3):312-8.