Molecular and Bioassay Examination of Neospora Caninum Infection in Bovine Aborted Fetuses in Khorasan Razavi Province, Iran

Document Type : Original Articles

Authors

1 Department of Pathobiology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad,

2 Department of Pathobiology, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad

3 Department of Pathobiology, Faculty of Veterinary Medicine, Ferdowsi university of Mashhad

10.32592/ARI.2025.80.3.843

Abstract

Neospora caninum plays a significant role in causing abortion and reproductive failure in dairy cattle. The majority of neosporosis-related abortions take place during the 5–6 months of gestation. Fetal death in the uterus, resorption, mummification, autolysis, stillbirth, birth with clinical symptoms, or being born clinically healthy but with chronic infection are all possible outcomes. The objective of the study was to identify N. caninum infection in aborted bovine fetuses through molecular analysis and mouse bioassay testing. From 2019 to 2022, 121 bovine aborted fetuses were collected from dairy farms in Khorasan Razavi province. The fetal brain samples were screened for detection of the parasite DNA by polymerase chain reaction assay (PCR). In addition, a portion of PCR-positive brain tissue was homogenized and inoculated into the peritoneum of five BALB/c mice. All mice were sacrificed six weeks post infection and examined using serology, microscopic, and PCR methods. If the mice's brain samples were PCR positive, the mouse bioassay test was repeated two times. The N. caninum DNA was detected in 19.8% of brain samples in bovine aborted fetuses. Among PCR-positive brain samples, only ten samples were suitable for mouse bioassay examination. All inoculated mice were seronegative without clinical signs after three times bioassays, although the brain samples of three mice groups were PCR-positive after repeated bioassays. The PCR results showed a moderate frequency of Neospora infection in aborted bovine fetuses. Furthermore, the isolates obtained in this study had low pathogenicity in BALB/c mice. It seems the isolates belong to an avirulent strain

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References

  1. Dubey J, Schares G, Ortega-Mora L. Epidemiology and control of neosporosis and Neospora caninum. Clinical microbiology reviews. 2007;20(2):323-67.
  2. Sadrebazzaz A, Habibi G, Haddadzadeh H, Ashrafi J. Evaluation of bovine abortion associated with Neospora caninum by different diagnostic techniques in Mashhad, Iran. Parasitology research. 2007;100(6):1257-60.
  3. Razmi GR, Zarea H, Naseri Z. A survey of Neospora caninum-associated bovine abortion in large dairy farms of Mashhad, Iran. Parasitology research. 2010;106(6):1419-23.
  4. Conraths J, Gottstein B. Neosporosis: general considerations. Protozoal abortion in farm ruminants, Wallingford: CAB international. 2007;425.
  5. Razmi GR, Zarae H, Nourbakhash M, Naseri Z. Estimating the rate of transplacental transmission of Neospora caninum to aborted fetuses in seropositive dams in Mashhad area, Iran. 2013.
  6. Nematollahi A, Moghaddam G, Jaafari R, Helan JA, Norouzi M. Study on outbreak of Neospora caninum-associated abortion in dairy cows in Tabriz (Northwest Iran) by serological, molecular and histopathologic methods. Asian Pacific Journal of Tropical Medicine. 2013;6(12):942-6.
  7. Wouda W. Neosporosis: Biology, transmission and clinical signs. Protozoal Abortion in Farm Ruminants Wallingford, England CAB international. 2007:46-53.
  8. Hajikolaei MH, Hamidinejat H, Ghorbanpoor M, Goraninejad S. Serological study of Neospora caninum infection in cattle from Ahvaz area, Iran. 2008.
  9. HR H. Serological study of Neospora caninum in pregnant dairy cattle in Tehran, Iran. 2010.
  10. Ansari-Lari M. Bovine neosporosis in Iran: a systematic review and meta-analysis. Preventive Veterinary Medicine. 2020;176:104913.
  11. Gharekhani J, Yakhchali M, Berahmat R. Neospora caninum infection in Iran (2004–2020): A review. Journal of Parasitic Diseases. 2020;44(4):671-86.
  12. Rafati N, Jaafarian M. The determination of prevalence of Neospora caninum in aborted fetuses in dairy cattle of Shahrekord area, Chahar Mahal Bakhtiari province, by Nested-PCR. Journal of Veterinary Laboratory Research. 2014;6(1):45-50.
  13. Salehi N, Gottstein B, Haddadzadeh H. Genetic diversity of bovine Neospora caninum determined by microsatellite markers. Parasitology international. 2015;64(5):357-61.
  14. Razmi GR, Maleki M, Farzaneh N, Talebkhan Garoussi M, Fallah A. First report of Neospora caninum-associated bovine abortion in Mashhad area, Iran. Parasitology Research. 2007;100(4):755-7.
  15. Salehi N, Haddadzadeh H, Ashrafihelan J, Shayan P, Sadrebazzaz A. Molecular and pathological study of bovine aborted fetuses and placenta from Neospora caninum infected dairy cattle. Iranian Journal of Parasitology. 2009;4(3):40-51.
  16. Kaveh A, Merat E, Samani S, Danandeh R, Nezad SS. Infectious Causes of Bovine Abortion in Qazvin Province, Iran. Archives of Razi Institute. 2017;72(4).
  17. Salehi N, Haddadzadeh H, Shayan P, Koohi MK. Isolation of Neospora caninum from an aborted fetus of seropositive cattle in Iran. 2012.
  18. Hoseini A, Merat E, Samani S, Nezhad SS, Danandeh R. Comparison of Neospora caninum infected tissues in aborted fetal bovine by PCR. 2018.
  19. Khani M, Arabkhazaeli F, Hosseini SD, Shayan P. Molecular detection of Neospora caninum in aborted fetuses of cattle farms in Arak. 2019.
  20. Amouei A, Sharif M, Sarvi S, Nejad RB, Aghayan SA, Hashemi-Soteh MB, et al. Aetiology of livestock fetal mortality in Mazandaran province, Iran. PeerJ. 2019;6:e5920.
  21. Müller N, Zimmermann V, Hentrich B, Gottstein B. Diagnosis of Neospora caninum and Toxoplasma gondii infection by PCR and DNA hybridization immunoassay. Journal of Clinical Microbiology. 1996;34(11):2850-2.
  22. Salehi B, Amouei A, Dodangeh S, Daryani A, Sarvi S, Safari-Kharyeki MR, et al. Molecular identification of Neospora caninum infection in aborted fetuses of sheep, cattle, and goats in Mazandaran Province, Northern Iran. Iranian Journal of Parasitology. 2021;16(3):483.
  23. Ansari-Lari M. Neospora caninum in aborted bovine fetuses in Iran: a systematic review and meta-analysis. Annals of Parasitology. 2021;67(3):357-66.
  24. Al-Qassab SE, Reichel MP, Ellis JT. On the biological and genetic diversity in Neospora caninum. Diversity. 2010;2(3):411-38.
  25. Calarco L, Barratt J, Ellis J. Genome wide identification of mutational hotspots in the apicomplexan parasite Neospora caninum and the implications for virulence. Genome biology and evolution. 2018;10(9):2417-31.
  26. Barr B, Anderson M, Blanchard P, Daft B, Kinde H, Conrad P. Bovine fetal encephalitis and myocarditis associated with protozoal infections. Veterinary Pathology. 1990;27(5):354-61.
  27. Conrad P, Barr B, Sverlow K, Anderson M, Daft B, Kinde H, et al. In vitro isolation and characterization of a Neospora sp. from aborted bovine foetuses. Parasitology. 1993;106(3):239-49.
  28. Lindsay D, Lenz S, Cole R, Dubey J, Blagburn B. Mouse model for central nervous system Neospora caninum infections. The Journal of parasitology. 1995:313-5.
  29. Miller C, Quinn H, Windsor P, Ellis J. Characterisation of the first Australian isolate of Neospora caninum from cattle. Australian Veterinary Journal. 2002;80(10):620-5.
  30. Rojo-Montejo S, Collantes-Fernández E, Regidor-Cerrillo J, Álvarez-García G, Marugan-Hernández V, Pedraza-Díaz S, et al. Isolation and characterization of a bovine isolate of Neospora caninum with low virulence. Veterinary parasitology. 2009;159(1):7-16.
  31. Dittrich RL, Regidor-Cerrillo J, Ortega-Mora LM, de Oliveira Koch M, Busch APB, Gonçalves KA, et al. Isolation of Neospora caninum from kidney and brain of a bovine foetus and molecular characterization in Brazil. Experimental parasitology. 2018;185:10-6.
  32. Dubey J, Shen S, Kwok O, Thulliez P. Toxoplasmosis in rats (Rattus norvegicus): congenital transmission to first and second generation offspring and isolation of Toxoplasma gondii from seronegative rats. Parasitology. 1997;115(1):9-14.
  33. Jenkins M, Parker C, Hill D, Pinckney R, Dyer R, Dubey J. Neospora caninum detected in feral rodents. Veterinary Parasitology. 2007;143(2):161-5.
Archives of Razi Institute
Volume 80, Issue 3
May and June 2025
Pages 843-848

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