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Home / Equine Research Bank / Parasit Vectors / Genetic variants of Anaplasma phagocytophilum from 14 equine...
Parasites & vectors2011; 4; 161; doi: 10.1186/1756-3305-4-161

Genetic variants of Anaplasma phagocytophilum from 14 equine granulocytic anaplasmosis cases.

Abstract: Equine Granulocytic Anaplasmosis (EGA) is caused by Anaplasma phagocytophilum, a tick-transmitted, obligate intracellular bacterium. In Europe, it is transmitted by Ixodes ricinus. A large number of genetic variants of A. phagocytophilum circulate in nature and have been found in ticks and different animals. Attempts have been made to assign certain genetic variants to certain host species or pathologies, but have not been successful so far. The purpose of this study was to investigate the causing agent A. phagocytophilum of 14 cases of EGA in naturally infected horses with molecular methods on the basis of 4 partial genes (16S rRNA, groEL, msp2, and msp4). Results: All DNA extracts of EDTA-blood samples of the horses gave bands of the correct nucleotide size in all four genotyping PCRs. Sequence analysis revealed 4 different variants in the partial 16S rRNA, groEL gene and msp2 genes, and 3 in the msp4 gene. One 16S rRNA gene variant involved in 11 of the 14 cases was identical to the "prototype" variant causing disease in humans in the amplified part [GenBank: U02521]. Phylogenetic analysis revealed as expected for the groEL gene that sequences from horses clustered separately from roe deer. Sequences of the partial msp2 gene from this study formed a separate cluster from ruminant variants in Europe and from all US variants. Conclusions: The results show that more than one variant of A. phagocytophilum seems to be involved in EGA in Germany. The comparative genetic analysis of the variants involved points towards different natural cycles in the epidemiology of A. phagocytophilum, possibly involving different reservoir hosts or host adaptation, rather than a strict species separation.
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Publication Date: 2011-08-16 PubMed ID: 21843364PubMed Central: PMC3170280DOI: 10.1186/1756-3305-4-161Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research focuses on exploring the genetic variants of Anaplasma phagocytophilum, a bacterium causing Equine Granulocytic Anaplasmosis (EGA) in horses, utilizing molecular methods on the basis of 4 partial genes.

Investigation of Anaplasma phagocytophilum

  • The research primarily centers on Anaplasma phagocytophilum, the obligate intracellular bacterium that is transmitted by ticks and results in the illness, EGA, within horses.
  • This bacterium circulates in variant forms across different animals and ticks in nature, but it is particularly transmitted by Ixodes ricinus in Europe.
  • Scientists previously attempted to associate specific variants of this bacterium to particular host species or pathologies, but they have not succeeded in their endeavors.

Research Purpose and Methods

  • The purpose of this study was to examine Anaplasma phagocytophilum, the causing agent of 14 EGA cases in horses that were naturally infected.
  • This examination was conducted using molecular methods based on 4 selected partial genes: 16S rRNA, groEL, msp2, and msp4.

Results

  • The DNA extracts of the blood samples from all the horses yielded bands that measured the correct nucleotide size in all four genotyping PCRs.
  • The sequence analysis revealed 4 different variants in the partial 16S rRNA, groEL gene, and msp2 genes, with the msp4 gene yielding 3 variants.
  • One variant of the 16S rRNA gene was found identical to the “prototype” variant that causes disease in humans and was present in 11 of the 14 cases.

Conclusions

  • The study concludes that more than one variant of the Anaplasma phagocytophilum bacterium seems to be involved in the occurrence of EGA in Germany.
  • The comparative genetic analysis indicates the existence of different natural cycles involving this bacterium, pointing towards possible implications of different reservoir hosts or host adaptation, rather than a strict species separation.

Cite This Article

APA
Silaghi C, Liebisch G, Pfister K. (2011). Genetic variants of Anaplasma phagocytophilum from 14 equine granulocytic anaplasmosis cases. Parasit Vectors, 4, 161. https://doi.org/10.1186/1756-3305-4-161

Publication

Parasites & vectors
ISSN: 1756-3305
NlmUniqueID: 101462774
Country: England
Language: English
Volume: 4
Pages: 161

Researcher Affiliations

Silaghi, Cornelia
  • Comparative Tropical Medicine and Parasitology, Faculty of Veterinary Medicine, Ludwig-Maximilians-Universität, Leopoldstr, 5, 80802 Munich, Germany. cornelia.silaghi@tropa.vetmed.uni-muenchen.de
Liebisch, Gabriele
    Pfister, Kurt

      MeSH Terms

      • Anaplasma phagocytophilum / classification
      • Anaplasma phagocytophilum / genetics
      • Anaplasma phagocytophilum / isolation & purification
      • Anaplasmosis / microbiology
      • Animals
      • Bacterial Outer Membrane Proteins / genetics
      • Chaperonin 60 / genetics
      • Cluster Analysis
      • DNA, Bacterial / chemistry
      • DNA, Bacterial / genetics
      • DNA, Ribosomal / chemistry
      • DNA, Ribosomal / genetics
      • Germany
      • Horse Diseases / microbiology
      • Horses
      • Molecular Sequence Data
      • Phylogeny
      • RNA, Ribosomal, 16S / genetics
      • Sequence Analysis, DNA

      References

      This article includes 46 references
      1. Woldehiwet Z. Anaplasma phagocytophilum in ruminants in Europe.. Ann N Y Acad Sci 2006;1078:446–460.
        doi: 10.1196/annals.1374.084pubmed: 17114753google scholar: lookup
      2. Dumler JS, Barbet AF, Bekker CP, Dasch GA, Palmer GH, Ray SC, Rikihisa Y, Rurangirwa FR. Reorganization of the 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 and designation of Ehrlichia equi and 'HGE agent' as subjective synonyms of Ehrlichia phagocytophila.. Int J Syst Evol Microbiol 2001;51:2145–2165.
        doi: 10.1099/00207713-51-6-2145pubmed: 11760958google scholar: lookup
      3. Von Loewenich FD, Baumgarten BU, Schröppel K, Geißdörfer W, Röllinghoff M, Bogdan C. High Diversity of ankA sequences of Anaplasma phagocytophilum among Ixodes ricinus ticks in Germany.. J Clin Microbiol 2003;41:2033–5040.
        doi: 10.1128/JCM.41.5.2033-2039.2003pmc: PMC262509pubmed: 14605135google scholar: lookup
      4. Zeman P, Pecha M. Segregation of genetic variants of Anaplasma phagocytophilum circulating among wild ruminants within a Bohemian forest (Czech Republic). Int J Med Microbiol 2008;298(Suppl 1):203–210.
      5. Scharf W, Schauer S, Freyburger F, Petrovec M, Schaarschmidt-Kiener D, Liebisch G, Runge M, Ganter M, Kehl A, Dumler JS, Garcia-Perez AL, Jensen J, Fingerle V, Meli ML, Ensser A, Stuen S, von Loewenich FD. Distinct host species correlate with Anaplasma phagocytophilum ankA gene clusters.. J Clin Microbiol 2011;49:790–796.
        doi: 10.1128/JCM.02051-10pmc: PMC3067700pubmed: 21177886google scholar: lookup
      6. Silaghi C, Scheuerle MC, Friche Passos LM, Thiel C, Pfister K. PCR detection of Anaplasma phagocytophilum in goat flocks in an area endemic for tick-borne fever in Switzerland.. Parasite 2011;18:57–62.
        pmc: PMC3671408pubmed: 21395206
      7. Silaghi C, Hamel D, Thiel C, Pfister K, Friche Passos LM, Rehbein S. Genetic variants of Anaplasma phagocytophilum in wild caprine and cervid ungulates from the Alps in Tyrol, Austria.. Vector Borne Zoonotic Dis 2011. in press.
        pubmed: 21323423
      8. Gribble DH. Equine ehrlichiosis.. J Am Vet Med Assoc 1969;155:462–469.
        pubmed: 5819585
      9. Büscher G, Gandras R, Apel G, Friedhoff KT. [The 1st case of ehrlichiosis in a horse in Germany (Brief report)]. Dtsch Tierarztl Wochenschr 1984;91:408–409.
        pubmed: 6397345
      10. Hermann M, Baumann D, Lutz H, Wild P. Erster diagnostizierter Fall von Equiner Ehrlichiose in der Schweiz.. Pferdeheilkunde 1985;1:247–250.
      11. Bjoersdorff A. Ehrlichia equi infection diagnosed in horses.. Svensk Veterinär Tidning 1990;42:357–360.
      12. Korbutiak E, Schneiders D. Equine Granulocytic Ehrlichiosis in the UK.. Vet Rec 1994;135:387–388.
        pubmed: 7831748
      13. Berrington A, Moats R, Lester S. A case of Ehrlichia equi in an adult horse in British Columbia.. Can Vet 1996;37:174–175.
        pmc: PMC1576643pubmed: 8681291
      14. Bermann F, Davoust B, Fournier PE, Brisou-Lapointe AV, Brouqui P. Ehrlichia equi (Anaplasma phagocytophila) infection in an adult horse in France.. Vet Rec 2002;150:787–788.
        doi: 10.1136/vr.150.25.787pubmed: 12135075google scholar: lookup
      15. Von Loewenich FD, Stumpf G, Baumgarten BU, Röllinghoff M, Dumler JS, Bogdan C. A case of equine granulocytic ehrlichiosis provides molecular evidence for the presence of pathogenic Anaplasma phagocytophilum (HGE agent) in Germany.. Eur J Clin Microbiol Infect Dis 2003;22:303–305.
        pubmed: 12740667
      16. Butler CM, Nijhof AM, Jongejan F, Van der Kolk JH. Anaplasma phagocytophilum infection in horses in the Netherlands.. Vet Rec 2008;162:216–218.
        doi: 10.1136/vr.162.7.216pubmed: 18281631google scholar: lookup
      17. Jahn P, Zeman P, Bezdekova B, Praskova I. Equine granulocytic anaplasmosis in the Czech Republic.. Vet Rec 2010;166:646–649.
        doi: 10.1136/vr.4852pubmed: 20495165google scholar: lookup
      18. Reubel GH, Kimsey RB, Barlough JE, Madigan JE. Experimental transmission of Ehrlichia equi to horses through naturally infected ticks (Ixodes pacificus) from Northern California.. J Clin Microbiol 1998;38:2131–2134.
        pmc: PMC105004pubmed: 9650983
      19. Franzén P, Aspan A, Egenvall A, Gunnarsson A, Åberg L, Pringle J. Acute Clinical, Hematologic, Serologic, and Polymerase Chain Reaction findings in horses experimentally infected with a European strain of Anaplasma phagocytophilum.. J Vet Intern Med 2005;19:232–239.
        pubmed: 15822569
      20. Madigan JE. Equine Ehrlichiosis.. Vet Clin North Am Equine Pract 1993;9:423–428.
        pubmed: 8358654
      21. Franzén P, Aspan A, Egenvall A, Gunnarsson A, Karlstam E, Pringle J. Molecular evidence for persistence of Anaplasma phagocytophilum in the absence of clinical abnormalities in horses after recovery from acute experimental infection.. J Vet Intern Med 2009;23:636–642.
        doi: 10.1111/j.1939-1676.2009.0317.xpubmed: 19645847google scholar: lookup
      22. Passamonti F, Veronesi F, Cappelli K, Capomaccio S, Coppola G, Marenzoni ML, Piergili FD, Verini Supplizi A, Coletti M. 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.2008.08.002pubmed: 18805584google scholar: lookup
      23. Hansen MGB, Christoffersen M, Thuesen LR, Petersen MR, Bojesen AM. Seroprevalence of Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum in Danish horses.. Act Vet Scand 2010;52:3.
        doi: 10.1186/1751-0147-52-3pmc: PMC2818635pubmed: 20082693google scholar: lookup
      24. Granquist EG, Bårdsen K, Bergström K, Stuen S. Variant - and individual dependent nature of persistent Anaplasma phagocytophilum infection.. Acta Vet Scand 2010;52:25.
        doi: 10.1186/1751-0147-52-25pmc: PMC2859769pubmed: 20398321google scholar: lookup
      25. Pusterla N, Lutz H, Braun U. Experimental infection of four horses with Ehrlichia phagocytophila.. Vet Rec 1998;143:303–305.
        doi: 10.1136/vr.143.11.303pubmed: 9789347google scholar: lookup
      26. Barlough JE, Madigan JE, DeRock E, Dumler JS, Bakken JS. Protection against Ehrlichia equi is conferred by prior infection with the Human Granulocytotropic Ehrlichia (HGE Agent). J Clin Microbiol 1995;33:3333–3334.
        pmc: PMC228702pubmed: 8586731
      27. Franzén P, Berg A-L, Aspan A, Gunnarson A, Pringle J. Death of a horse infected experimentally with Anaplasma phagocytophilum.. Vet Rec 2007;160:122–125.
        doi: 10.1136/vr.160.4.122pubmed: 17259454google scholar: lookup
      28. Alberti A, Zobba R, Chessa B, Addis MF, Sparagano O, Parpaglia MLP, Cupeddu T, Pintori G, Pittau M. Equine and Canine Anaplasma phagocytophilum strains isolated on the island of Sardinia (Italy) are phylogenetically related to pathogenic strains from the United States.. Appl Env Microbiol 2005;71:6418–6422.
        doi: 10.1128/AEM.71.10.6418-6422.2005pmc: PMC1265917pubmed: 16204571google scholar: lookup
      29. Silaghi C, Gilles J, Höhle M, Fingerle V, Just FT, Pfister K. Anaplasma phagocytophilum infection in Ixodes ricinus, Bavaria, Germany.. Emerg Infect Dis 2008;14:972–974.
        doi: 10.3201/eid1406.071095pmc: PMC2600299pubmed: 18507918google scholar: lookup
      30. 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 2005;43:1309–1317.
        doi: 10.1128/JCM.43.3.1309-1317.2005pmc: PMC1081214pubmed: 15750101google scholar: lookup
      31. Johansson K-E, Pettersson B, Uhlén M, Gunnarsson A, Malmquist M, Olsson E. Identification of the causative agent of granulocytic ehrlichiosis in Swedish dogs and horses by direct solid phase sequencing of PCR products from the 16S rRNA gene.. Res Vet Sci 1995;58:109–112.
        doi: 10.1016/0034-5288(95)90061-6pubmed: 7539151google scholar: lookup
      32. Zeman P, Jahn P. An entropy-optimized multilocus approach for characterizing the strains of Anaplasma phagocytophilum infecting horses in the Czech Republic.. J Med Microbiol 2009;58:423–429.
        doi: 10.1099/jmm.0.007831-0pubmed: 19273636google scholar: lookup
      33. Leonard S. Doctoral Thesis.. Ludwig-Maxmilians-Universtät München, Institute of Comparative Tropical Medicine and Parasitology; 2005.
      34. Schauer S. Doctoral Thesis.. Albert-Ludwigs-Universität Freiburg im Breisgau, Institute for Medical Microbiology and Hygiene; 2008.
      35. Parola P, Davoust B, Raoult D. Tick- and flea-borne rickettsial emerging zoonoses.. Vet Res 2005;36:469–92.
        doi: 10.1051/vetres:2005004pubmed: 15845235google scholar: lookup
      36. Massung RF, Courtney JW, Hiratzka SL, Pitzer VE, Smith G, Dryden RL. Anaplasma phagocytophilum in white-tailed deer.. Emerg Infect Dis 2005;11:1604–6.
        pmc: PMC3366735pubmed: 16318705
      37. Foley J, Nieto NC, Madigan J, Sykes J. Possible differential host tropism in Anaplasma phagocytophilum strains in the Western United States.. Ann N Y Acad Sci 2008;1149:94–97.
        doi: 10.1196/annals.1428.066pubmed: 19120182google scholar: lookup
      38. Pusterla N, Huder JB, Feige K, Lutz H. Identification of a Granulocytic Ehrlichia Strain isolated from a horse in Switzerland and comparison with other rickettsiae of the Ehrlichia phagocytophila genogroup.. J Clin Microbiol 1998;36:2035–2037.
        pmc: PMC104973pubmed: 9650957
      39. Granquist EG, Stuen S, Lundgren AM, Braten M, Barbet AF. Outer Membrane Protein sequence variation in lambs experimentally infected with Anaplasma phagocytophilum.. Infect Immun 2008;76:120–126.
        doi: 10.1128/IAI.01206-07pmc: PMC2223638pubmed: 17967854google scholar: lookup
      40. Teglas MB, Foley J. Differences in the transmissibility of two Anaplasma phagocytophilum strains by the North American tick vector species, Ixodes pacificus and Ixodes scapularis (Acari: Ixodidae). Exp Appl Acarol 2006;38:47–58.
        doi: 10.1007/s10493-005-5293-5pubmed: 16550334google scholar: lookup
      41. Courtney JW, Kostelnik LM, Nordin SZ, Massung RF. Multiplex Real-Time PCR for detection of Anaplasma phagocytophilum and Borrelia burgdorferi.. J Clin Microbiol 2004;42:3164–3168.
        doi: 10.1128/JCM.42.7.3164-3168.2004pmc: PMC446246pubmed: 15243077google scholar: lookup
      42. Massung RF, Slater K, Owens JH, Nicholson WL, Mather TN, Solberg VB, Olson JG. Nested PCR assay for detection of granulocytic ehrlichiae.. J Clin Microbiol 1998;36:1090–1095.
        pmc: PMC104695pubmed: 9542943
      43. Lin Q, Rikihisa Y, Massung RF, Woldehiwet Z, Falco RC. Polymorphism and Transcription at the p44-1/p44-18 Genomic Locus in Anaplasma phagocytophilum Strains from Diverse Geographic Regions.. Infect Immun 2004;72:5574–5581.
        doi: 10.1128/IAI.72.10.5574-5581.2004pmc: PMC517535pubmed: 15385454google scholar: lookup
      44. Bown KJ, Lambin X, Ogden NH, Petrovec M, Shaw SE, Woldehiwet Z, Birtles RJ. High-resolution genetic fingerprinting of European strains of Anaplasma phagocytophilum by use of multilocus variable-number tandem-repeat analysis.. J Clin Microbiol 2007;45:1771–1776.
        doi: 10.1128/JCM.00365-07pmc: PMC1933113pubmed: 17442796google scholar: lookup
      45. Chenna R, Sugawara H, Koike T, Lopez R, Gibson TJ, Higgins DG, Thompson JD. Multiple sequence alignment with the Clustal series of programs.. Nucleic Acids Res 2003;31:3497–500.
        doi: 10.1093/nar/gkg500pmc: PMC168907pubmed: 12824352google scholar: lookup
      46. Felsenstein J. PHYLIP - phylogeny interference package (version 3.2). Cladistics 1989;5:164–166.
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