Skip to main content

Advertisement

Log in

Neospora caninum NC-6 Argentina induces fetopathy in both serologically positive and negative experimentally inoculated pregnant dams

  • Original Paper
  • Published:
Parasitology Research Aims and scope Submit manuscript

Abstract

Neospora caninum infection is a major cause of abortion in cattle. The objectives of this study were to genetically characterize the N. caninum NC-6 Argentina isolate using a multilocus microsatellite analysis approach and to study its biological behavior by experimental inoculations into seronegative and seropositive pregnant cattle, evaluating the humoral and cellular immune response elicited and the occurrence of transplacental transmission and fetopathy. Pregnant cows (65 days of gestation) seropositive and seronegative to N. caninum were intravenously inoculated with tachyzoites of the NC-6 Argentina N. caninum strain and slaughtered at 108 ± 2 days of gestation. Serum samples were analyzed for N. caninum antibodies by indirect fluorescent antibody test. The cellular immune response was analyzed by detection of gamma interferon (γIFN) production in blood cells. Tissue samples from dams, fetuses, and placental cotyledons were processed by histopathological and immunohistochemical techniques and examined for N. caninum DNA by PCR. Positive DNA samples were further analyzed by multilocus microsatellite typing for N. caninum. Inoculated animals had significantly higher N. caninum antibody titers and γIFN production than control animals. One seropositive inoculated cow aborted, one seronegative cow had a non-viable fetus, and the remaining fetuses from the experimentally inoculated dams had histopathologic lesions. The PCR was positive in 3/4 fetuses from seronegative inoculated cows and in 2/3 fetuses from seropositive inoculated cows. Multilocus microsatellite analysis revealed that the N. caninum DNA present in fetuses and placentas had an identical pattern to NC-6 Argentina strain. The NC-6 Argentina strain proved to be able to cross the placenta and to induce fetopathy in both the seropositive and seronegative dams.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Al-Qassab S, Reichel MP, Ellis J (2010) A second generation multiplex PCR for typing strains of Neospora caninum using six DNA targets. Mol Cell Probes 24:20–26

    Article  PubMed  CAS  Google Scholar 

  • Al-Qassab S, Reichel MP, Ivens A, Ellis JT (2009) Genetic diversity amongst isolates of Neospora caninum, and the development of a multiplex assay for the detection of distinct strains. Mol Cell Probes 23:132–139

    Article  PubMed  CAS  Google Scholar 

  • Anderson ML, Andrianarivo AG, Conrad PA (2000) Neosporosis in cattle. Anim Reprod Sci 60–61:417–431

    Article  PubMed  Google Scholar 

  • Anderson ML, Blanchard PC, Barr BC, Dubey JP, Hoffman RL, Conrad PA (1991) Neospora like protozoan infection as a major cause of abortion in California dairy cattle. J Am Vet Med Assoc 198:241–244

    PubMed  CAS  Google Scholar 

  • Barr BC, Rowe JD, Sverlow KW, BonDurant RH, Ardans AA, Oliver MN, Conrad PA (1994) Experimental reproduction of bovine fetal Neospora infection and death with a bovine Neospora isolate. J Vet Diagn Invest 6:207–215

    Article  PubMed  CAS  Google Scholar 

  • Basso W, Schares S, Minke L, Bärwald A, Maksimov A, Peters M, Schulze C, Müller M, Conraths FJ, Schares G (2010) Microsatellite typing and avidity analysis suggest a common source of infection in herds with epidemic Neospora caninum-associated bovine abortion. Vet Parasitol 173:24–31

    Article  PubMed  CAS  Google Scholar 

  • Basso W, Venturini L, Venturini MC, Hill DE, Kwok OCH, Shen SK, Dubey JP (2001) First isolation of Neospora caninum from the feces of a naturally infected dog. J Parasitol 87:612–618

    PubMed  CAS  Google Scholar 

  • Basso W, Herrmann DC, Conraths FJ, Pantchev N, Vrhovec MG, Schares G (2009a) First isolation of Neospora caninum from the faeces of a dog from Portugal. Vet Parasitol 159:162–166

    Article  PubMed  CAS  Google Scholar 

  • Basso W, Schares S, Bärwald A, Herrmann DC, Conraths FJ, Pantchev N, Vrhovec MG, Schares G (2009b) Molecular comparison of Neospora caninum oocyst isolates from naturally infected dogs with cell culture-derived tachyzoites of the same isolates using nested polymerase chain reaction to amplify microsatellite markers. Vet Parasitol 160:43–50

    Article  PubMed  CAS  Google Scholar 

  • Beck HP, Blake D, Darde ML, Felger I, Pedraza-Diaz S, Regidor-Cerrillo J, Gomez-Bautista M, Ortega-Mora LM, Putignani L, Shiels B, Tait A, Weir W (2009) Molecular approaches to diversity of populations of apicomplexan parasites. Int J Parasitol 39:175–189

    Article  PubMed  CAS  Google Scholar 

  • Björkman C, Uggla A (1999) Serological diagnosis of Neospora caninum infection. Int J Parasitol 29:1497–1507

    Article  PubMed  Google Scholar 

  • Campero CM, Anderson ML, Conosciuto G, Odriozola H, Bretschneider G, Poso MA (1998) Neospora caninum-associated abortion in a dairy herd in Argentina. Vet Record 143:228–229

    Article  CAS  Google Scholar 

  • Campero C, Moore D, Odeon A, Cipolla A, Odriozola E (2003) Aetiology of bovine abortion in Argentina. Vet Res Commun 27:359–369

    Article  PubMed  CAS  Google Scholar 

  • Davison HC, Otter A, Trees AJ (1999) Estimation of vertical and horizontal transmission parameters of Neospora caninum infection in dairy cattle. Int J Parasitol 29:1683–1689

    Article  PubMed  CAS  Google Scholar 

  • Dubey JP (1999) Neosporosis in cattle: biology and economic impact. J Am Vet Med Assoc 214:1160–1163

    PubMed  CAS  Google Scholar 

  • Dubey JP (2003a) Neosporosis in cattle. J Parasitol 89(Suppl):S42–S56

    Google Scholar 

  • Dubey JP (2003b) Review of Neospora caninum and neosporosis in animals. Korean J Parasitol 60:1–16

    Article  Google Scholar 

  • Dubey JP, Hattel AL, Lindsay DS, Topper MJ (1988) Neonatal Neospora caninum infection in dogs: isolation of the causative agent and experimental transmission. J Am Vet Med Assoc 193:1259–1263

    PubMed  CAS  Google Scholar 

  • Dubey JP, Schares G (2006) Diagnosis of bovine neosporosis. Vet Parasitol 140:1–34

    Article  PubMed  CAS  Google Scholar 

  • Gondim LF, McAllister MM, Pitt WC, Zemlicka DE (2004) Coyotes (Canis latrans) are definitive hosts of Neospora caninum. Int J Parasitol 34:159–161

    Article  PubMed  Google Scholar 

  • Gottstein B, Hentrich R, Wyss B, Thür A, Busato A, Stärk KDC, Müller N (1998) Molecular and immunodiagnostic investigations on bovine neosporosis in Switzerland. Int J Parasitol 28:679–691

    Article  PubMed  CAS  Google Scholar 

  • Hall CA, Reichel MP, Ellis JT (2005) Neospora Abortion in dairy cattle: diagnosis, mode of transmission and control. Vet Parasitol 128:231–241

    Article  PubMed  CAS  Google Scholar 

  • Innes EA, Andrianarivo AG, Björkman C, Williams DJL, Conrad PA (2002) Immune responses to Neospora caninum and prospects for vaccination. Trends Parasitol 18:497–504

    Article  PubMed  CAS  Google Scholar 

  • Innes EA, Wright S, Bartley P, Maley S, Macaldowie C, Esteban-Redondo I, Buxton D (2005) The host–parasite relationship in bovine neosporosis. Vet Immunol Immunopathol 108:29–36

    Article  PubMed  Google Scholar 

  • Jenkins M, Baszler T, Björkman C, Schaes G, Williams D (2002) Diagnosis and seroepidemiology of Neospora caninum-associated bovine abortion. Int J Parasitol 32:631–636

    Article  PubMed  Google Scholar 

  • Macaldowie C, Maley SW, Wright S, Bartley P, Esteban-Redondo I, Buxton D, Innes EA (2004) Placental pathology associated with fetal death in cattle inoculated with Neospora caninum by two different routes in early pregnancy. J Comp Path 131:142–156

    Article  PubMed  CAS  Google Scholar 

  • Maley SW, Buxton D, Macaldowie CN, Anderson IE, Wright SE, Bartley PM, Esteban-Redondo I, Hamilton CM, Storsety AK, Innes EA (2006) Characterization of the immune response in the placenta of cattle experimentally infected with Neospora caninum in early gestation. J Comp Path 135:130–141

    Article  PubMed  CAS  Google Scholar 

  • Mc Allister M, Dubey JP, Lindsay DS, Jolley WR, Wills RA, Mc Guire AM (1998) Dogs are definitive hosts of Neospora caninum. J Parasitol 28:1473–1478

    CAS  Google Scholar 

  • Moré G, Basso W, Bacigalupe D, Venturini MC, Venturini L (2008) Diagnosis of Sarcocystis cruzi, Neospora caninum, and Toxoplasma gondii infections in cattle. Parasitol Res 102:671–675

    Article  PubMed  Google Scholar 

  • Moré G, Bacigalupe D, Basso W, Rambeaud M, Beltrame F, Ramirez B, Venturini MC, Venturini L (2009) Frequency of horizontal and vertical transmission for Sarcocystis cruzi and Neospora caninum in dairy cattle. Vet Parasitol 160:51–54

    Article  PubMed  Google Scholar 

  • Paré J, Thurmond MC, Hietala SK (1996) Congenital Neospora caninum infection in dairy cattle and associated calfhood mortality. Can J Vet Res 60:133–139

    PubMed  Google Scholar 

  • Pedraza-Diaz S, Marugan-Hernandez V, Collantes-Fernandez E, Regidor-Cerrillo J, Rojo-Montejo S, Gomez-Bautista M, Ortega-Mora LM (2009) Microsatellite markers for the molecular characterization of Neospora caninum: application to clinical samples. Vet Parasitol 166:38–46

    Article  PubMed  CAS  Google Scholar 

  • Regidor-Cerrillo J, Pedraza-Diaz S, Gomez-Bautista M, Ortega-Mora LM (2006) Multilocus microsatellite analysis reveals extensive genetic diversity in Neospora caninum. J Parasitol 92:517–524

    Article  PubMed  CAS  Google Scholar 

  • Regidor-Cerrillo J, Gomez-Bautista M, Pereira-Bueno J, Aduriz G, Navarro-Lozano V, Risco-Castillo V, Fernandez-Garcia A, Pedraza-Diaz S, Ortega-Mora LM (2008) Isolation and genetic characterization of Neospora caninum from asymptomatic calves in Spain. Parasitology 135:1651–1659

    Article  PubMed  CAS  Google Scholar 

  • Rojo-Montejo S, Collantes-Fernandez E, Regidor-Cerrillo J, Alvarez-Garcia G, Marugan-Hernandez V, Pedraza-Diaz S, Blanco-Murcia J, Prenafeta A, Ortega-Mora LM (2009) Isolation and characterization of a bovine isolate of Neospora caninum with low virulence. Vet Parasitol 159:7–16

    Article  PubMed  Google Scholar 

  • SENASA Resolución N° 115 (1999) Plan Nacional de Control y Erradicación de la Brucelosis y Tuberculosis. Secretaría de Agricultura, Ganadería, Pesca y Alimentación, Argentina

  • Thilsted JP, Dubey JP (1989) Neosporosis-like abortion in a herd of dairy cattle. J Vet Diagn Invest 1:205–209

    Article  PubMed  CAS  Google Scholar 

  • Thurmond MC, Hietala SK (1997) Effect of Neospora caninum infection on milk production in first-lactation dairy cows. J Am Vet Med Assoc 210:672–674

    PubMed  CAS  Google Scholar 

  • Thurmond MC, Hietala SK (1996) Culling associated with Neospora caninum infection in dairy cows. Am J Vet Res 57:1559–1562

    PubMed  CAS  Google Scholar 

  • Venturini MC, Venturini L, Bacigalupe D, Machuca M, Echaide I, Basso W, Unzaga JM, Di Lorenzo C, Guglielmone A, Jenkins MC, Dubey JP (1999) Neospora caninum infections in bovine fetuses and dairy cows with abortions in Argentina. Int J Parasitol 29:1705–1708

    Article  PubMed  CAS  Google Scholar 

  • Yamage M, Flechtner O, Gottstein B (1996) Neospora caninum: specific oligonucleotide primers for the detection of brain “cyst” DNA of experimentally infected nude mice by the polymerase chain reaction (PCR). J Parasitol 82:272–279

    Article  PubMed  CAS  Google Scholar 

  • Williams DJL, Guy CS, McGarry JM, Guy F, Tasker L, Smith RF, MacEachern K, Cripps PJ, Kelly DF, Trees AJ (2000) Neospora caninum-associated abortion in cattle: the time of experimentally induced parasitaemia during gestation determines foetal survival. Parasitology 121:347–358

    Article  PubMed  Google Scholar 

  • Williams DJL, Guy CS, Smith RF, Ellis J, Björkman C, Reichel MP, Trees AJ (2007) Immunization of cattle with live tachyzoites of Neospora caninum confers protection against fetal death. Infect Immun 75:1343–1348

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Isidoro Ercoli and Susann Schares for their excellent technical assistance and Magdalena Rambeaud for language revision of the paper.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to D. Bacigalupe.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bacigalupe, D., Basso, W., Caspe, S.G. et al. Neospora caninum NC-6 Argentina induces fetopathy in both serologically positive and negative experimentally inoculated pregnant dams. Parasitol Res 112, 2585–2592 (2013). https://doi.org/10.1007/s00436-013-3424-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00436-013-3424-1

Keywords

Navigation