Immunodiagnosis and Molecular Characterization of Sarcocystis Species in Cattle
KrishiKosh
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Title |
Immunodiagnosis and Molecular Characterization of Sarcocystis Species in Cattle
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Creator |
Mamatha, G.S.
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Contributor |
Placid E. D'Souza
Krishnappa, G. Suryanarayana, V.V.S. Yathiraj, S. Suguna Rao |
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Subject |
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Description |
Ph.D. Thesis
Sarcocystis bovicanis and Sarcocystis bovifelis species were encountered in this study in cattle. Immunodiagnosis of Sarcocystosis was undertaken by Enzyme linked immunosorbent assay and Enzyme immuno transfer blot by using soluble extract of S.bovicanis and S.bovifelis bradyzoites. The sensitivity and specificity of ELISA was found to be 76.4 and 81.8% for S.bovicanis and 66.6 and 71.4% for S.bovifelis respectively. Out of 300 cattle screened, 48.66 and 15.33% were positive for S.bovicanis and S.bovifelis infection respectively by ELISA. The sensitivity and specificity of EITB was found to be 87.5 and 81.8% for S.bovicanis and 93.5 and 87.5% for S.bovifelis respectively. EITB detected 58.3 and 29.6% of S.bovicanis and S.bovifelis infection respectively. The protein profile of soluble extract of S.bovicanis, S.bovifelis and host tissue by SDSPAGE revealed a total of 10, 11 and 9 polypeptides in range of 170 to 12kDa, 100 to 15kDa and 80 to 11kDa for S.bovicanis, S.bovifelis and host tissue respectively. The presence of three common bands between these two species by SDS-PAGE indicated that both the Sarcocystis species are closely related. EITB could detect a higher percentage of infected animals. The polypeptides detected on Western blots using homologous hyperimmune serum revealed polypeptides in range of 100 to 12 kDa for S.bovicanis and 100 to 14 kDa for S.bovifelis. However none of the polypeptides reacted with host tissue. Molecular characterization of the two species was conducted by RAPD-PCR with the four primers. Primer 1 amplified 1.3 and 1.2 kb fragments for S.bovicanis and 1.1, 1.0. 0.85 and 0.75 kb for S.bovifelis. DNA fragments of 0.4, 0.6, 0.7 and 1.2, 1.3 and 1.5 kb were amplified for S.bovicanis and S.bovifelis respectively. Primer 3 in S.bovicanis amplified 0.8 and 0.5 kb fragments and in the case of S.bovifelis 1.0 and 1.1 kb were amplified. S.bovicanis DNA amplified only one fragment of size 0.5 kb whereas S.bovifelis DNA amplified 0.7, 0.9 and 1.0 kb for primer 4. The RAPD-PCR indicated that the primer 3 and 4 are valuable diagnostic probes for differentiation of S.bovicanis and S.bovifelis species. |
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Date |
2016-08-02T12:48:38Z
2016-08-02T12:48:38Z 2006-09-12 |
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Type |
Thesis
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Identifier |
http://krishikosh.egranth.ac.in/handle/1/70524
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Language |
en
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Format |
application/pdf
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Publisher |
Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar
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