Hostname: page-component-76fb5796d-skm99 Total loading time: 0 Render date: 2024-04-27T00:17:38.864Z Has data issue: false hasContentIssue false

Detection of Neospora caninum in aborted bovine fetuses and dam blood samples by nested PCR and ELISA and seroprevalence in Beijing and Tianjin, China

Published online by Cambridge University Press:  07 August 2009

L. YAO
Affiliation:
Key Laboratory of Zoonosis of Ministry of Agriculture and National animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
N. YANG
Affiliation:
Key Laboratory of Zoonosis of Ministry of Agriculture and National animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
Q. LIU*
Affiliation:
Key Laboratory of Zoonosis of Ministry of Agriculture and National animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
M. WANG
Affiliation:
Key Laboratory of Zoonosis of Ministry of Agriculture and National animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
W. ZHANG
Affiliation:
Key Laboratory of Zoonosis of Ministry of Agriculture and National animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
W. F. QIAN
Affiliation:
Key Laboratory of Zoonosis of Ministry of Agriculture and National animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
Y. F. HU
Affiliation:
Key Laboratory of Zoonosis of Ministry of Agriculture and National animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
J. DING
Affiliation:
Key Laboratory of Zoonosis of Ministry of Agriculture and National animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
*
*Corresponding author: Tel: +86 10 62734496. Fax: +86 10 62732804. E-mail: qunliu@cau.edu.cn

Summary

Neospora caninum infection is a significant cause of abortion in cattle. We investigated the tissue distribution of N. caninum in aborted bovine fetuses and dam blood samples by a nested PCR assay, and compared the nested PCR with ELISA in the diagnosis of N. caninum infection. In total, 26 aborted fetuses and 813 blood samples were collected from 8 dairy herds in Beijing (n=212) and Tianjin (n=601), China. Fifteen fetuses (57·7%) were tested N. caninum-positive by the nested PCR. N. caninum DNA was detected from the brain of 52%, kidneys of 22%, skeletal muscle of 18%, and heart of 4% of the aborted fetuses. The PCR-positive cases (55%, 11/20) were higher than seropositive cows (40%, 8/20) in a subset of 20 fetuses, but the PCR results of blood samples of the 20 cows were all negative. The seroprevalence of the 813 samples was 15·5% (43·4% of samples from Beijing, 5·7% of samples from Tianjin), compared to the PCR-positive blood samples of 0·9%. Our study showed that the nested PCR is a valuable diagnostic tool for the primary diagnosis of N. caninum in aborted fetuses, while ELISA is the preferred assay for testing blood samples collected from cows. The two assays are complementary in determining whether abortions are associated with N. caninum infection in cattle.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2009

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Anderson, M. L., Andrianarivo, A. G. and Conrad, P. A. (2000). Neosporosis in cattle. Animal Reproduction Science 60–61, 417431.CrossRefGoogle ScholarPubMed
Baszler, T. V., Gay, L. J., Long, M. T. and Mathison, B. A. (1999). Detection by PCR of Neospora caninum in fetal tissues from spontaneous bovine abortions. Journal of Clinical Microbiology 37, 40594064.CrossRefGoogle ScholarPubMed
Buxton, D., Maley, S. W., Wright, S., Thomson, K. M., Rae, A. G. and Innes, E. A. (1998). The pathogenesis of experimental neosporosis in pregnant sheep. Journal of Comparative Pathology 118, 267279.CrossRefGoogle ScholarPubMed
Buxton, D., Wright, S., Maley, S. W., Rae, A. G., Lunden, A. and Innes, E. A. (2001). Immunity to experimental neosporosis in pregnant sheep. Parasite Immunology 23, 8591.CrossRefGoogle ScholarPubMed
Caetano-Da-Silva, A., Ferre, I., Aduriz, G., Alvarez-Garcia, G., Del-Pozo, I., Atxaerandio, R., Regidor-Cerrillo, J., Ugarte-Garagalza, C. and Ortega-Mora, L. M. (2004). Neospora caninum infection in breeder bulls: seroprevalence and comparison of serological methods used for diagnosis. Veterinary Parasitology 124, 1924.CrossRefGoogle ScholarPubMed
Collantes-Fernandez, E., Rodriguez-Bertos, A., Arnaiz-Seco, I., Moreno, B., Aduriz, G. and Ortega-Mora, L. M. (2006). Influence of the stage of pregnancy on Neospora caninum distribution, parasite loads and lesions in aborted bovine foetuses. Theriogenology 65, 629641.CrossRefGoogle ScholarPubMed
Crawshaw, W. M. and Brocklehurst, S. (2003). Abortion epidemic in a dairy herd associated with horizontally transmitted Neospora caninum infection. Veterinary Record 152, 201206.CrossRefGoogle Scholar
Dubey, J. P. (2003). Review of Neospora caninum and neosporosis in animals. Korean Journal of Parasitology 41, 116.CrossRefGoogle ScholarPubMed
Dubey, J. P., Carpenter, J. L., Speer, C. A., Topper, M. J. and Uggla, A. (1988). Newly recognized fatal protozoan disease of dogs. Journal of the American Veterinary Medical Association 192, 12691285.Google ScholarPubMed
Dubey, J. P. and Lindsay, D. S. (1996). A review of Neospora caninum and neosporosis. Veterinary Parasitology 67, 159.CrossRefGoogle ScholarPubMed
Dubey, J. P. and Schares, G. (2006). Diagnosis of bovine neosporosis. Veterinary Parasitology 140, 134.CrossRefGoogle ScholarPubMed
Ferre, I., Aduriz, G., Del-Pozo, I., Regidor-Cerrillo, J., Atxaerandio, R., Collantes-Fernandez, E., Hurtado, A., Ugarte-Garagalza, C. and Ortega-Mora, L. M. (2005). Detection of Neospora caninum in the semen and blood of naturally infected bulls. Theriogenology 63, 15041518.CrossRefGoogle ScholarPubMed
Gilbert, I., Bissonnette, N., Boissonneault, G., Vallee, M. and Robert, C. (2007). A molecular analysis of the population of mRNA in bovine spermatozoa. Reproduction 133, 10731086.CrossRefGoogle ScholarPubMed
Gondim, L. F., McAllister, M. M., Pitt, W. C. and Zemlicka, D. E. (2004). Coyotes (Canis latrans) are definitive hosts of Neospora caninum. International Journal for Parasitology 34, 159161.CrossRefGoogle ScholarPubMed
Gottstein, B., Hentrich, B., Wyss, R., Thur, B., Busato, A., Stark, K. D. and Muller, N. (1998). Molecular and immunodiagnostic investigations on bovine neosporosis in Switzerland. International Journal for Parasitology 28, 679691.CrossRefGoogle ScholarPubMed
Ho, M. S., Barr, B. C., Rowe, J. D., Anderson, M. L., Sverlow, K. W., Packham, A., Marsh, A. E. and Conrad, P. A. (1997). Detection of Neospora sp. from infected bovine tissues by PCR and probe hybridization. Journal of Parasitology 83, 508514.CrossRefGoogle ScholarPubMed
Huang, C. C., Ting, L. J., Shiau, J. R., Chen, M. C. and Ooi, H. K. (2004). An abortion storm in cattle associated with neosporosis in Taiwan. Journal of Veterinary Medical Science 66, 465467.CrossRefGoogle ScholarPubMed
Kaufmann, H., Yamage, M., Roditi, I., Dobbelaere, D., Dubey, J. P., Holmdahl, O. J., Trees, A. and Gottstein, B. (1996). Discrimination of Neospora caninum from Toxoplasma gondii and other apicomplexan parasites by hybridization and PCR. Molecular and Cellular Probes 10, 289297.CrossRefGoogle ScholarPubMed
Liao, M., Xuan, X., Huang, X., Shirafuji, H., Fukumoto, S., Hirata, H., Suzuki, H. and Fujisaki, K. (2005). Identification and characterization of cross-reactive antigens from Neospora caninum and Toxoplasma gondii. Parasitology 130, 481488.CrossRefGoogle ScholarPubMed
Liu, J., Cai, J. Z., Zhang, W., Liu, Q., Chen, D., Han, J. P. and Liu, Q. R. (2008). Seroepidemiology of Neospora caninum and Toxoplasma gondii infection in yaks (Bos grunniens) in Qinghai, China. Veterinary Parasitology 152, 330332.CrossRefGoogle ScholarPubMed
Liu, J., Yu, J., Wang, M., Liu, Q., Zhang, W., Deng, C. and Ding, J. (2007). Serodiagnosis of Neospora caninum infection in cattle using a recombinant tNcSRS2 protein-based ELISA. Veterinary Parasitology 143, 358363.CrossRefGoogle ScholarPubMed
McAllister, M. M., Dubey, J. P., Lindsay, D. S., Jolley, W. R., Wills, R. A. and McGuire, A. M. (1998). Dogs are definitive hosts of Neospora caninum. International Journal for Parasitology 28, 14731478.CrossRefGoogle ScholarPubMed
McInnes, L. M., Ryan, U. M., O'handley, R., Sager, H., Forshaw, D. and Palmer, D. G. (2006). Diagnostic significance of Neospora caninum DNA detected by PCR in cattle serum. Veterinary Parasitology 142, 207213.CrossRefGoogle ScholarPubMed
Moore, D. P. (2005). Neosporosis in South America. Veterinary Parasitology 127, 8797.CrossRefGoogle ScholarPubMed
Moskwa, B., Cabaj, W., Pastusiak, K. and Bien, J. (2003). The suitability of milk in detection of Neospora caninum infection in cows. Acta Parasitologica 48, 138141.Google Scholar
Muller, N., Zimmermann, V., Hentrich, B. and Gottstein, B. (1996). Diagnosis of Neospora caninum and Toxoplasma gondii infection by PCR and DNA hybridization immunoassay. Journal of Clinical Microbiology 34, 28502852.CrossRefGoogle ScholarPubMed
Okeoma, C. M., Williamson, N. B., Pomroy, W. E., Stowell, K. M. and Gillespie, L. (2004). The use of PCR to detect Neospora caninum DNA in the blood of naturally infected cows. Veterinary Parasitology 122, 307315.CrossRefGoogle ScholarPubMed
Ortega-Mora, L. M., Ferre, I., Del-Pozo, I., Caetano-Da-Silva, A., Collantes-Fernandez, E., Regidor-Cerrillo, J., Ugarte-Garagalza, C. and Aduriz, G. (2003). Detection of Neospora caninum in semen of bulls. Veterinary Parasitology 117, 301308.CrossRefGoogle ScholarPubMed
Peters, M., Wagner, F. and Schares, G. (2000). Canine neosporosis: clinical and pathological findings and first isolation of Neospora caninum in Germany. Parasitology Research 86, 17.CrossRefGoogle ScholarPubMed
Schatzberg, S. J., Haley, N. J., Barr, S. C., Delahunta, A., Olby, N., Munana, K. and Sharp, N. J. (2003). Use of a multiplex polymerase chain reaction assay in the antemortem diagnosis of toxoplasmosis and neosporosis in the central nervous system of cats and dogs. American Journal of Veterinary Research 64, 15071513.CrossRefGoogle ScholarPubMed
Van Maanen, C., Wouda, W., Schares, G., Von Blumroder, D., Conraths, F. J., Norton, R., Williams, D. J., Esteban-Redondo, I., Innes, E. A., Mattsson, J. G., Bjorkman, C., Fernandez-Garcia, A., Ortega-Mora, L. M., Muller, N., Sager, H. and Hemphill, A. (2004). An interlaboratory comparison of immunohistochemistry and PCR methods for detection of Neospora caninum in bovine foetal tissues. Veterinary Parasitology 126, 351364.CrossRefGoogle ScholarPubMed
Yamage, M., Flechtner, O. and 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). Journal of Parasitology 82, 272279.CrossRefGoogle ScholarPubMed
Yu, J., Xia, Z., Liu, Q., Liu, J., Ding, J. and Zhang, W. (2007). Seroepidemiology of Neospora caninum and Toxoplasma gondii in cattle and water buffaloes (Bubalus bubalis) in the People's Republic of China. Veterinary Parasitology 143, 7985.CrossRefGoogle ScholarPubMed
Zhang, W., Deng, C., Liu, Q., Liu, J., Wang, M., Tian, K. G., Yu, X. L. and Hu, D. M. (2007). First identification of Neospora caninum infection in aborted bovine foetuses in China. Veterinary Parasitology 149, 7276.CrossRefGoogle ScholarPubMed