Original Article

Molecular Identification of Neospora caninum Infection in Aborted Fetuses of Sheep, Cattle, and Goats in Mazandaran Province, Northern Iran


Background: We aimed to identify Neospora caninum DNA in the brain samples of aborted fetuses of cattle, goats, and sheep in Mazandaran, northern Iran, using PCR.

Methods: In total, 133 aborted fetuses (51 sheep, 78 cattle, and 4 goats) were randomly collected from different stages of gestation in various regions of Mazandaran, Iran, from Mar 2016 to May 2017. The DNA was extracted from all the brain samples using phenol chloroform isoamyl alcohol instructions. The Nc-5 gene was used for the detection of N. caninum DNA by nested-PCR assay.

Results: The detection of N. caninum DNA was confirmed by the observation of a 227 bp band in 24 samples of 133 aborted fetuses (18.1%). The highest prevalence rate of N. caninum was detected in the cattle (20.5%) followed by the sheep (15.6%); however, no positive cases were reported in the goats. The highest and lowest prevalence rates of the infection were reported as 23.8% and 8.6% in Qaemshahr, and Behshahr, respectively. The prevalence rate of infection (32%) in the early gestational period was higher than those in the middle (15%) and late (3.8%) gestational periods.

Conclusion: The obtained data of the present study indicated that N. caninum infection may partly be responsible for abortion and economic loss in livestock farming in Mazandaran Province.

1. Dubey J, Schares G. Neosporosis in animals the last five years. Vet Parasitol. 2011;180(1-2): 90-108.
2. Shaapan RM. The common zoonotic protozoal diseases causing abortion. J Parasit Dis.2016;40(4): 1116-1129.
3. Dubey JP, Lindsay DS. A review of Neospora caninum and neosporosis. Vet Parasitol. 1996;67(1-2): 1-59.
4. Dubey JP. Review of Neospora caninum and neosporosis in animals. Korean J Parasitol. 2003;41(1):1-16.
5. Anastasia D, Elias P, Nikolaos P, et al. Toxoplasma gondii and Neospora caninum seroprevalence in dairy sheep and goats mixed stock farming.Vet Parasitol. 2013;198(3-4):387-90.
6. Pescador C, Corbellini L, Oliveira E, et al. Histopathological and immunohistochemical aspects of Neospora caninum diagnosis in bovine aborted fetuses Vet Parasitol.2007;150(1-2): 159-63.
7. Uzêda R, Schares G, Ortega-Mora LM, et al. Combination of monoclonal antibodies improves immunohistochemical diagnosis of Neospora caninum. Vet Parasitol. 2013;197(3-4): 477-86.
8. Tian A-L, Elsheikha HM, Zhou D-H, et al. A novel recombinase polymerase amplification (RPA) assay for the rapid isothermal detection of Neospora caninum in aborted bovine fetuses. Vet Parasitol. 2018;258(15): 24-29.
9. Habibi GR, Hashemi-Fesharki R, Sadrebazzaz A, et al. Seminested PCR for Diagnosis of Neospora caninum Infection in Cattle. Archives of Razi Institute. 2005;59(2): 55-64.
10. Hajikolaei MRH, Goraninejad S, Hamidinejat H, et al. Occurrence of Neospora caninum antibodies in water buffaloes (Bubalus bubalis) from the south-western region of Iran. Bulletin- Veterinary Institute in Pulawy. 2007;51(2): 233-235.
11. Fard SRN, Khalili M, Aminzadeh AJVA. Prevalence of antibodies to Neospora caninum in cattle in Kerman province, South East Iran. Vet Arhiv. 2008;78(3): 253-259.
12. Nematollahi A, Jaafari R, Moghaddam Gh. Seroprevalence of Neospora caninum infection in dairy cattle in Tabriz, Northwest Iran. Iran J Parasitol. 2011;6(4): 95-98.
13. Kamali A, Seifi HA, Movassaghi AR, et al. Histopathological and molecular study of Neospora caninum infection in bovine aborted fetuses. Asian Pacific J Trop Biomed. 2014;4(12):990-994.
14. Hughes J, Williams R, Morley E, et al . The prevalence of Neospora caninum and co-infection with Toxoplasma gondii by PCR analysis in naturally occurring mammal populations. Parasitology. 2006; 132(Pt 1):29-36.
15. Müller N, Zimmermann V, Hentrich B, et al. Diagnosis of Neospora caninum and Toxoplasma gondii infection by PCR and DNA hybridization immunoassay. J Clin Microbiol. 1996;34(11): 2850-2852.
16. Amouei A, Sharif M, Sarvi S, et al. Aetiology of livestock fetal mortality in Mazandaran province, Iran. Peer J. 2019; 6: e5920.
17. Reichel MP, Ayanegui-Alcérreca MA, Gondim LF, et al. What is the global economic impact of Neospora caninum in cattle–the billion dollar question. Int J Parasitol. 2013;43(2): 133-42.
18. Yao L, Yang N, Liu Q, et al . Detection of Neospora caninum in aborted bovine fetuses and dam blood samples by nested PCR and ELISA and seroprevalence in Beijing and Tianjin, China. Parasitology. 2009;136(11):1251-6.
19. Asadpour R, Jafari-Joozani R, Salehi N. Detection of Neospora caninum in ovine abortion in Iran. J Parasit Dis. 2013;37(1): 105-9.
20. Sasani F, Javanbakht J, Seifori P, et al. Neospora caninum as causative agent of ovine encephalitis in Iran. Pathol Discov.2013;1(1): 5.
21. 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.
22. Sadrebazzaz A, Habibi G, Haddadzadeh H, et al. Evaluation of bovine abortion associated with Neospora caninum by different diagnostic techniques in Mashhad, Iran. Parasitol Res. 2007. 100(6):1257-60.
23. Santos SL, de Souza Costa K, Gondim LQ, et al. Investigation of Neospora caninum, Hammondia sp., and Toxoplasma gondii in tissues from slaughtered beef cattle in Bahia, Brazil. Parasitol Res. 2010;106(2): 457-61.
24. Masala G, Porcu R, Daga C, et al. Detection of pathogens in ovine and caprine abortion samples from Sardinia, Italy, by PCR. J Vet Diagn Invest. 2007;19(1): 96-8.
25. Moreno B, Collantes-Fernández E, Villa A, et al. Occurrence of Neospora caninum and Toxoplasma gondii infections in ovine and caprine abortions. Vet Parasitol. 2012;187(1-2): 312-8.
26. Gharekhani J. Seroprevalence of Neospora caninum and Toxoplasma gondii infections in aborted cattle in Hamedan, Iran. J Adv Vet Anim Res. 2014;1(2): 32-35.
27. Al-Qassab SE, Reichel MP, Ellis JT. On the biological and genetic diversity in Neospora caninum. Diversity. 2010 Mar;2(3):411-438.
28. Pan Y, Jansen G, Duffield T, et al. Genetic susceptibility to Neospora caninum infection in Holstein cattle in Ontario. J Dairy Sci. 2004;87(11): 3967-75.
29. Amdouni Y, Rjeibi M, Awadi S, et al. First detection and molecular identification of Neospora caninum from naturally infected cattle and sheep in North Africa. Transbound Emerg Dis. 2018;65(4): 976-982.
30. Eleni C, Crotti S, Manuali E, et al . Detection of Neospora caninum in an aborted goat foetus. Vet Parasitol. 2004;123(3-4): 271-4.
31. De Meerschman F, Speybroeck N, Berkvens D, et al. Fetal infection with Neospora caninum in dairy and beef cattle in Belgium. Theriogenology. 2002;58(5): 933-45.
32. Sager H, Fischer I, Furrer K, et al. A Swiss case–control study to assess Neospora caninum-associated bovine abortions by PCR, histopathology and serology. Vet Parasitol. 2001;102(1-2): 1-15.
33. Innes EA, Andrianarivo AG, Björkman C, et al. Immune responses to Neospora caninum and prospects for vaccination. Trends Parasitol. 2002;18(11): 497-504.
IssueVol 16 No 3 (2021) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijpa.v16i3.7102
Neospora caninum Ruminants’ aborted fetuses Nested-PCR; Iran

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How to Cite
Salehi B, Amouei A, Dodangeh S, Daryani A, Sarvi S, Safari-Kharyeki MR, Salehi S, Hosseini SA, Hosseininejad Z. Molecular Identification of Neospora caninum Infection in Aborted Fetuses of Sheep, Cattle, and Goats in Mazandaran Province, Northern Iran. Iran J Parasitol. 2021;16(3):483-489.