Iranian Journal of Parasitology 2014. 9(3):394-401.

Isolation, Identification, and Pathogenicity of Neospora caninum China Yanbian strain.
Li-Jun Jia, Shou-Fa Zhang, Ming-Ming Liu, Nian-Chao Qian, Huan-Ping Guo


Background: The aim of the study was to provide a point of reference to study the Neospora caninum infections in China.

Methods: Genome DNA was extracted from the brains of aborted fetuses and specific PCR was performed with N. caninum Nc5-targeted specific primers. Fetal bovine brain tissues were homogenized and continuously cultured in Vero cells with double antibodies. The medium was replaced at 2-d intervals and the state of cells was observed.

Results: A 608 bp Nc5 gene band was detected by PCR amplification. After sequencing,the sequence of the sample shared 99.5% homology with GenBank(AF061249). Brain homogenates were continuously cultured in Vero cells for 34 d and N. caninum was found. The results of IFAT and Nc5 gene-based PCR detection were N. caninum-positive, and the parasite was tentatively named N. caninum China Yanbian strain. BABL/c mice were inoculated with the separated parasites and showed clinical symptoms of ataxia and limb paralysis after 12 d. Only 3 mice survived.The blood of dying mice and the hearts, livers, spleens, lungs, kidneys, andbrains of dead mice were collected aseptically. The Nc5 gene-based PCR showed that N. caninum may exist in brains, livers, and spleen. Based on immunohistochemical observations, we showed that N. caninum tachyzoites existed in the brains and livers.

Conclusion: We have successfully isolated bovine-specific N. caninum strain from brain tissues of aborted cattle in the China Yanbian region. This isolated strain has a strong infectious ability towards BABL/c mice.


Bovine; China; Isolation; Neospora caninum

Full Text:



Davison HC, Trees AJ, Guy F, Otter A, Holt JJ, Simpson VR, Jeffrey M. Isolation of bovine Neospora in Britain. Vet Rec. 1997; 141:607.

Yamane I, Kokuho T, Shimura K, Eto M, Shibahara T, Haritani M, Ouchi Y, Sverlow K, Conrad PA. In vitro isolation and characterization of a bovine Neospora species in Japan. Rec Vet Sci. 1997; 63: 77-80.

Bjerkås I, Mohn SF, Presthus J. Unidentified cystforming sporozoon causing encephalomyelitis and myositis in dogs. Z Parasitenkd. 1984; 70:271-274.

Dubey JP, Carpenter JL, Speer CA, Topper MJ, Uggla A. Newly recognized falal protozoan disease of dogs. J Am Vet Med Assoc. 1988; 192:1269-1285.

Dubey JP. Neosporosis in cattle; biology and economic impact. J Am Vet Med Assoc. 1999; 214:1160-1163.

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

Stenlund S, Kindahl H, Magnusson U, Uggla A, Björkman C. Serum antibody profile and reproductive performance during two consecutive pregnancies of cows naturally infected with Neospora caninum. Vet Parasitol. 1999; 85:227-234.

Jenkins M, Baszler T, Bjorkman C, Schares G, Williams D. Diagnosis and seroepidemiology of Neospora caninum-associated boine abortion. Int J Parasitol. 2002; 32:631-636.

Cramer G, Kelton D, Duffield TF, Hobson JC, Lissemore K, Hietala SK, Peregrine AS. Neospora caninum serostatus and culling of Holstein cattle. J Am Vet Med Assoc. 2002; 221:1165-1168.

Cañón-Franco WA, Bergamaschi DP, Labruna MB, Camargo LM, Souza SL, Silva JC, Pinter A, Dubey JP, Gennari SM. Prevalence of antibodies to Neospora caninum in dogs from Amazon, Brazil. Vet Parasitol. 2003; 115:71-74.

Chao W, Ma L, Li W, Zhang Z, Li D, Han W. Serological investigation of Neospora caninum infection in dairy cows in Geermu county of Qinghai province. Chinese J Vety Sci Technol. 2005; 35:1012-1014.

Liu Q, Li B, Qi C, Zhu Y. The first report on the serological test of Neosporosis in dairy cows. Chinese J Vet Med. 2003; 39:8-9.

Bayin C, Hao J, Wang C. Serologic tests with the recombinant NcSRS1 antigen on the cattle infected with Neospora caninum in Xinjiang partial areas. Chinese J Vet Parasitol. 2008; 16:10-12.

Zhang S, Ding D, Ju Y, Jia L. The comparison of colloidal gold immunochromatography and ELISA used in the detection of Neospora caninum sera antibody. Chinese J Vet Med. 2008; 44:54-55.

Wang C, Li Z, Zhai Y, Ni H, Chen A, Chen J, Wang N. Development and preliminary application of indirect ELISA diagnostic method for Neospora caninum in dairy cattle. Chinese J Preven Vet Med. 2009; 31:145-149.

Deng C, Zhang W, Liu Q, Hu D, Yu X, Liu J. Identification of Neospora caninum in aborted fetus of dairy cow in China. Chinese Vet Sci. 2007; 37:16-19.

Ding D, Jia L, Tian W, Liang W, Zhang X, Zhang S. Isolation and identification of Jilin strain Neospora caninum. Chin Vet Sci. 2009; 29:423-425.

Langoni H, Matteucci G, Medici B, Camossi LG, Richini-Pereira VB, Silva RC. Detection and molecular analysis of Toxoplasma gondii and Neospora caninum from dogs with neurological disorders. Rev Soc Bras Med Trop. 2012; 45:365-368.

Meerburg BG, De Craeye S, Dierick K, Kijlstra A. Neospora caninum and Toxoplasma gondii in brain tissue of feral rodents and insectivores caught on farms in the Netherlands. Vet Parasitol. 2012; 184:317-320.

Hughes JM, Williams RH, Morley EK, Cook DA, Terry RS, Murphy RG, Smith JE, Hide G. The prevalence of Neospora caninum and co-infection with Toxoplasma gondii by PCR analysis in naturally occurring mammal populations. Parasitology. 2006; 132:29-36.

Ghalmi F, China B, Kaidi R, Daube G, Losson B. Detection of Neospora caninum in dog organs using real time PCR systems. Vet Parasitol. 2008; 155:161-167.

Truppel JH, Montiani-Ferreira F, Lange RR, Vilani RG, Reifur L, Boerger W, da Costa-Ribeiro MC, Thomaz-Soccol V. Detection of Neospora caninum DNA in capybaras and phylogenetic analysis. Parasitol Int. 2010; 59:376-379.

Lindsay DS, Dubey JP, Duncan RB. Confirmation that the dog is a definitive host for Neospora caninum. 1999; Vet Parasitol 82:327-333.

Peters M, Lütkefels E, Heckeroth AR, Schares G. Immunohistochemical and ultrastructural evidence for Neospora caninum tissue cysts in skeletal muscles of naturally infected dogs and cattle. Int J Parasitol. 2001; 31:1144-1148.

De Marez T, Liddell S, Dubey JP, Jenkins MC, Gasbarre L. Oral infection of calves with Neospora caninum oocysts from dogs: humoral and cellular immune responses. Int J Parasitol. 1999; 29:1647-1657.

Pereira-Bueno J, Quintanilla-Gozalo A, Pérez- Pérez V, Espi-Felgueroso A, Alvarez-García G, Collantes-Fernández E, Ortega-Mora LM. Evaluation by different diagnostic techniques of bovine abortion associated with Neospora caninum in Spain. Vet Parasitol. 2003; 111: 143-152.

Sharifdini M, Mohebali M, Keshavarz H, Hosseininejad M, Hajjaran H, Akhoundi B, Foroushani AR, Zarei Z, Charehdar S. Neospora caninum and Leishmania infantum Co-Infection in Domestic Dogs (Canis familiaris) in Meshkin- Shahr District, Northwestern Iran. Iran J Arthropod- Borne Dis, 2011, 5(2): 60–68.

Čobádiová A, Reiterová K, Derdáková M, Špilovská S, Turčeková L, Hviščová I, Hisira V. Toxoplasma gondii, Neospora caninum and tick-transmitted bacterium Anaplasma phagocytophilum infections in one selected goat farm in Slovakia. Acta Parasitol. 2013 ,58(4):541-546.

Okeoma CM, Williamson NB, Pomroy WE, Stowell KM, Gillespie L. The use of PCR to detect Neospora caninum DNA in the blood of naturally infected cows. Vet Parasitol. 2004; 122: 307-315.

Maley SW, Buxton D, Rae AG, Wright SE, Schock A, Bartley PM, Esteban-Redondo I, Swales C, Hamilton CM, Sales J, Innes EA. The pathogenesis of neosporosis in pregnant cattle: inoculation at mid-gestation. J Comp Pathol. 2003; 129:186-195.

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

Deng C, Zhang W, Liu Q, Liu J, Ding J. BALB/c Mouse Model of neosporosis and the development process with infection of Neospora caninum. Scientia Agricultura Sinica. 2009; 42:1123-1128.


  • There are currently no refbacks.

Creative Commons Attribution-NonCommercial 3.0

This work is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.