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Immunodiagnosis and Molecular Characterization of Sarcocystis Species in Cattle

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Title Immunodiagnosis and Molecular Characterization of Sarcocystis Species in Cattle
 
Creator Mamatha, G.S.
 
Contributor Placid E. D'Souza
Krishnappa, G.
Suryanarayana, V.V.S.
Yathiraj, S.
Suguna Rao
 
Subject ---
 
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.
 
Date 2016-08-02T12:48:38Z
2016-08-02T12:48:38Z
2006-09-12
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/70524
 
Language en
 
Format application/pdf
 
Publisher Karnataka Veterinary, Animal and Fisheries Sciences University, Bidar