PDF herunterladen
Tierarztl Prax Ausg K Kleintiere Heimtiere 2012; 40(06): 399-403
DOI: 10.1055/s-0038-1623671
Original Article
Schattauer GmbH

Analysis of the culture-derived soluble Babesia canis canis antigens derived from the Polish strains of the parasites

Analyse des löslichen parasitären Antigens aus In-vitro-Kulturen der endemischen polnischen Stämme von Babesia canis canis
L. Adaszek
1   Department of Epizootiology and Infectious Diseases, Faculty of Veterinary Medicine, University of Life Sciences, Lublin, Poland
,
A. Puchalski
2   Sub-Department of Veterinary Prevention, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, Lublin, Poland
,
M. Dec
2   Sub-Department of Veterinary Prevention, Institute of Biological Bases of Animal Diseases, Faculty of Veterinary Medicine, University of Life Sciences, Lublin, Poland
,
S. Winiarczyk
1   Department of Epizootiology and Infectious Diseases, Faculty of Veterinary Medicine, University of Life Sciences, Lublin, Poland
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received: 26. Februar 2012

Accepted after revision: 23. Mai 2012

Publikationsdatum:
06. Januar 2018 (online)

    Lizenzen und Reprints

Summary

Objective, material and methods: The aim of this study was to analyse the protein fractions of the soluble parasitic antigen (SPA) from in vitro cultures of the native Polish strains of Babesia canis canis and to determine their immunogenicity through Western blotting using the sera of dogs vaccinated with this antigen.

Results: Polyacrylamide gel electrophoresis revealed 21 protein fractions with molecular weights from 12 to 205 kDa. The most intense reaction in Western blotting was observed between the serum antibodies of the SPA-vaccinated dogs and the fraction with the molecular weight of 52 kDa.

Conclusion: Detailed studies on the composition of SPA of Babesia canis canis and reactivity of its individual protein fractions can be a starting point for the development of subunit vaccines against babesiosis. Using a preparation with only some electrophoretic fractions of SPA in the production of vaccines would allow to avoid putting an unnecessary protein burden in the vaccine which could cause side effects.

Zusammenfassung

Ziel, Material und Methoden: Ziel dieser Studie war, aus In-vitro- Kulturen der endemischen polnischen Stämme von Babesia canis canis einzelne Proteinfraktionen des löslichen parasitären Antigens (soluble parasitic antigen, SPA) zu analysieren und deren Immunogenität im Western Blot mit den Seren der mit diesem Antigen geimpften Hunde zu bestimmen.

Ergebnisse: Durch Polyacrylamid-Gelelektrophorese ließen sich 21 Proteinfraktionen mit den Molekulargewichten von 12 bis 205 kDa isolieren. Die intensivste Reaktion in Western Blot ergab sich zwischen den Serumantikörpern der SPA-geimpften Hunde und der Fraktion mit einem Molekulargewichten von 52 kDa.

Schlussfolgerung: Detaillierte Untersuchungen zur Zusammensetzung des SPA von Babesia canis canis und zur Reaktivität der einzelnen Proteinfraktionen können als Grundlage für die Entwicklung von Subunit-Impfstoffen gegen die Babesiose des Hundes dienen. Bei Verwendung einzelner SPA- Fraktionen lassen sich belastende Fremdproteine im Impfstoff und daraus möglicherweise resultierende Nebenwirkungen bei geimpften Tieren reduzieren.

Key words

Babesia canis canis - Western blotting - SPA - dogs

Schlüsselwörter

Babesia canis canis - Western Blot - SPA - Hund
 

  • References

  • 1 Adaszek L, Winiarczyk S. Babesiosis in dogs – still an actual problem. Wiad Parazytol 2008; 54: 109-115.
    PubMedGoogle Scholar
  • 2 Adaszek L, Winiarczyk S. Molecular characterization of Babesia canis canis isolates from naturally infected dogs in Poland. Vet Parasitol 2008; 152: 235-241.
    CrossrefPubMedGoogle Scholar
  • 3 Adaszek L, Winiarczyk S, Skrzypczak M. The clinical course of babesiosis in 76 dogs infected with protozoa parasites Babesia canis canis. Pol J Vet Sci 2009; 12: 81-87.
    PubMedGoogle Scholar
  • 4 Adaszek L, Winiarczyk S. In vitro cultivation of Babesia canis canis parasites isolated from dogs in Poland. Parasitol Res 2011; 108: 1303-1307.
    CrossrefPubMedGoogle Scholar
  • 5 Adaszek L, Martinez AC, Winiarczyk S. The factors affecting the distribution of babesiosis in dogs in Poland. Vet Parasitol 2011; 181: 160-165.
    CrossrefPubMedGoogle Scholar
  • 6 Azzar G, Radisson J, Got R. Characterization and purification of culture-derived soluble glycoproteins of Babesia canis . Parasitol Res 1990; 76: 578-580.
    CrossrefPubMedGoogle Scholar
  • 7 Cardoso L, Costa A, Tuna J, Vieira L, Eyal O, Yisaschar-Mekuzas Y, Baneth G. Babesia canis canis and Babesia canis vogeli infections in dogs from northern Portugal. Vet Parasitol 2008; 156: 199-204.
    CrossrefPubMedGoogle Scholar
  • 8 Carret C, Walas F, Carcy B, Grande N, Précigout É, Moubri K, Schetters T, Gorenflot A. Babesia canis canis, Babesia canis vogeli, Babesia canis rossi: Differentiation of three subspecies by restriction fragment length polymorphism analysis on amplified small subunit ribosomal RNA genes. J Eukaryot Micro- biol 1999; 46: 298-303.
    PubMedGoogle Scholar
  • 9 Day MJ. The immunopathology of canine vector-borne diseases. Parasit Vectors 2011; 13 (04) 48.
    PubMedGoogle Scholar
  • 10 Goodger BV, Wright IG, Mahoney DF, McKenna RV. Babesia bovis (Argentina): studies on the composition and location of antigen associated with in fected erythrocytes. Int J Parasitol 1980; 10: 33-36.
    CrossrefPubMedGoogle Scholar
  • 11 Gothe R, Wegerdt S. Die Babesiosen des Hundes in Deutschland: epidemio logische Fallanalysen. Tierärztl Prax 1991; 19: 170-173.
    PubMedGoogle Scholar
  • 12 Gravely SM, Smith RD, Erp EE, Cantó GJ, Aikawa M, Osorno BM, Ristic M. Bovine babesiosis: partial purification and characterization of blood culture- derived Babesia bovis . Int J Parasitol 1979; 09: 591-598.
    CrossrefPubMedGoogle Scholar
  • 13 Hauschild S, Shayan P, Schein E. Characterization and comparison of merozoite antigens of different Babesia canis isolates by serological and immuno- logical investigations. Parasitol Res 1995; 81: 638-642.
    CrossrefPubMedGoogle Scholar
  • 14 James MA, Levy MG, Ristic M. Isolation and partial characterization of culture-derived soluble Babesia bovis antigens. Infect Immun 1981; 31: 358-361.
    PubMedGoogle Scholar
  • 15 Kahl LP, Anders RF, Rodwell BJ, Timms P, Mitchell GF. Variable and common antigens of Babesia bovis parasites differing in strain and virulence. J Immunol 1982; 129: 1700-1705.
    PubMedGoogle Scholar
  • 16 Kubelová M, Tkadlec E, Bednár M, Roubalová E, Siroký P. West-to-east differences of Babesia canis canis prevalence in Dermacentor reticulatus ticks in Slovakia. Vet Parasitol 2011; 180: 191-196.
    CrossrefPubMedGoogle Scholar
  • 17 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685.
    CrossrefPubMedGoogle Scholar
  • 18 Laurent N, l’Etoile M. Process for the culture of Babesia canis . Application to the preparation of antigens and vaccine against pyroplasmosis. Patent no. US 4,777,036, 1988
    PubMedGoogle Scholar
  • 19 Milczak A, Riha T, Abramowicz B, Madej E. Hemostatic disorders during the course of canine babesiosis. Medyna Wet 2004; 60: 1067-1070.
    PubMedGoogle Scholar
  • 20 Nussenzweig RS. The application of monoclonal antibodies to studies of malaria. In: Properties of the monoclonal antibodies produced by hybridome technology and their application to the study of diseases. Houba V, Chart SH. eds. National University of Singapore UNDP/World Bank/WHO; Geneva: 1982: 63-69.
    Google Scholar
  • 21 Schetters TP, Kleuskens JA, Scholtes NC, Pasman JW, Bos HJ. Vaccination of dogs against Babesia canis infection using antigens from culture supernatants with emphasis on clinical babesiosis. Vet Parasitol 1994; 52: 219-233.
    CrossrefPubMedGoogle Scholar
  • 22 Schetters TP, Montenegro-James S. Vaccines against babesiosis using soluble parasite antigens. Parasitol Today 1995; 11: 456-462.
    CrossrefPubMedGoogle Scholar
  • 23 Schetters TP, Scholtes NC, Kleuskens JA, Bos HJ. Not peripheral parasitaemia but the level of soluble parasite antigen in plasma correlates with vaccine ef ficacy against Babesia canis . Parasite Immunol 1996; 18: 1-6.
    CrossrefPubMedGoogle Scholar
  • 24 Schetters TP, Kleuskens JA, Scholtes NC, Pasman JW, Goovaerts D. Vaccination of dogs against Babesia canis infection. Vet Parasitol 1997; 73: 35-41.
    CrossrefPubMedGoogle Scholar
  • 25 Schetters TP, Kleuskens J, Carcy B, Gorenflot A, Vermeulen A. Vaccination against large Babesia species from dogs. Parasitologia 2007; 49 (Suppl. 01) 13-17.
    PubMedGoogle Scholar
  • 26 Schetters TP, Strydom T, Crafford D, Kleuskens JA, van de Crommert J, Vermeulen AN. Immunity against Babesia rossi infection in dogs vaccinated with antigens from culture supernatants. Vet Parasitol 2007; 144: 10-19.
    CrossrefPubMedGoogle Scholar
  • 27 Schetters TP, Moubri K, Cooke BM. Comparison of Babesia rossi and Babesia canis isolates with emphasis on effects of vaccination with soluble parasite antigens: a review. J S Afr Vet Assoc 2009; 80: 75-78.
    CrossrefPubMedGoogle Scholar
  • 28 Solano-Gallego L, Trotta M, Carli E, Carcy B, Caldin M, Furlanello T. Babesia canis canis and Babesia canis vogeli clinicopathological findings and DNA detection by means of PCR-RFLP in blood from Italian dogs suspected of tick-borne disease. Vet Parasitol 2008; 157: 211-221.
    CrossrefPubMedGoogle Scholar
  • 29 Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76: 4350-4354.
    CrossrefPubMedGoogle Scholar
  • 30 Yoshida N, Nussenzweig RS, Potocnjak P, Nussenzweig V, Aikawa M. Hybridoma produces protective antibodies directed against the sporozoite stage of malaria parasite. Science 1980; 207: 71-73.
    CrossrefPubMedGoogle Scholar
  • 31 Zahler M, Schein E, Rinder H, Gothe R. Characteristic genotypes discriminate between Babesia canis isolates of differing vector specificity and pathogenicity in dogs. Parasitol Res 1998; 84: 544-548.
    CrossrefPubMedGoogle Scholar
  • 32 Zygner W, Rapacka G, Gójska-Zygner O, Dlugosz E, Wedrychowicz H. Biochemical abnormalities observed in serum of dogs infected with large Babesia in Warsaw (Poland). Pol J Vet Sci 2007; 10: 245-253.
    PubMedGoogle Scholar