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ThesisItem Open Access MOLECULAR CHARACTERIZATION OF SARCOCYSTIS spp. ISOLATED FROM PIGS IN COASTAL DISTRICTS OF ANDHRA PRADESH, INDIA(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2022-01) MADHURI, GATTA; Jyothi Sree, Ch (MAJOR); Sreedevi, C; Sudhakar, KThe present study was carried out to determine the prevalence, morphological and molecular characteristics of Sarcocystis species in pigs in coastal Andhra Pradesh by microscopy and polymerase chain reaction using tissue samples collected from 163 slaughtered pigs for a period of one year from January 2021 to December 2021. Data regarding age, sex, breed and area of the animals were noted. Microscopic examination of tissue samples collected from different predilection sites such as heart, tongue, oesophagus, intercostals and diaphragmatic pillar muscles from each of 163 slaughtered pigs by intact cyst isolation and pepsin digestion method revealed sarcocysts in 6 (3.68%) and 20 (12.26%) pigs samples, respectively. Tissue samples positive for sarcocysts were fixed in 10% formalin for histopathological examination. Sarcocysts were microscopic with a smooth and thin cyst wall and were filled with banana-shaped bradyzoites. No thick walled cysts were identified. The morphological features of the cystic stage were suggestive of S. suihominis. Histopathological examination of affected muscle tissues revealed mild leucocytic infiltration, myositis and hyaline degeneration along with fatty degeneration. DNA was isolated from each pig tissue sample separately and was separately used to amplify partial segment of 18S rRNA gene of Sarcocystis spp. with the estimated amplicon size of 915 bp. Out of 163 samples, 34 (20.85 %) samples were found to be positive for Sarcocystis species. Subsequent digestion of PCR amplicons with SspI, two different patterns were visualized on electrophoresis, one with 915 bp (S. suihominis) and other with 650 bp and 265 bp (S. meischeriana). Of the 34 positive pigs, 31 (91.2%) were found to be positive for S. suihominis infection, one (2.9%) was found to be positive for S. meischeriana and two pigs (5.9%) were positive for mixed species infection with S. suihominis and S. meischeriana. Sequences of S. suihominis obtained from pigs demonstrated 90-100% identity with the Indian isolates of S. suihominis as single monophyletic group along with Italian isolate of S. suihominis. The prevalence was high in Krishna (36.36%) followed by Guntur (22.22%), Visakhapatnam (20.0%), East Godavari (18.18%), Srikakulam (15.83%), Nellore (14.28%), West Godavari, (8.33%), Prakasam (7.69%) and Vijayanagaram (7.14%) districts of coastal Andhra Pradesh. Age and breed influenced the prevalence of Sarcocystis infection being more prevalent in Non descriptive pigs (P<0.01) and adults (P<0.05). No significant difference was observed between the prevalence of male and female pigs. Overall, the prevalence of S. suihominis (20.24%; 33/163) was more compared to that of S. miescheriana (1.84%; 3/163) in the study area possessing public health risk.
