DIAGNOSIS OF BOVINE SARCOCYSTOSIS BY IMMUNOFLUORESCENT ANTIBODY TECHNIQUE
| dc.contributor.advisor | UDAYA KUMAR, M(MAJOR) | |
| dc.contributor.advisor | CHENGALVA RAYULU, V | |
| dc.contributor.advisor | NARASIMHA REDDY, Y | |
| dc.contributor.advisor | ANAND KUMAR, A | |
| dc.contributor.author | DASMA BAI, BANOTHU | |
| dc.date.accessioned | 2018-11-14T10:48:02Z | |
| dc.date.available | 2018-11-14T10:48:02Z | |
| dc.date.issued | 2012-03 | |
| dc.description | THESES | en_US |
| dc.description.abstract | ABSTRACT: Sarcocystis species are widely prevalent in man and animals, causing significant impact on animal and public health throughout the world. A laboratory standardized immunofluorescent antibody technique was used to study the seroprevalence of bovine sarcocystosis and the efficacy of same was compared with the traditional diagnostic methods like macroscopic, microscopic (squash and pepsin HCl acid muscle digestion). Histopathological changes and viability of bradyzoites in the affected esophageal muscles were also studied in the present investigation. The gross examination of oesophagi revealed creamy white colored thin walled macrocysts of Sarcocystis spp appearing in different shapes (fusiform, elliptical, ovoidal and globular etc) and sizes ranging from 2.0-18.0 x 1.0-5.0 mm with an average size of 10.47 + 0.295 x 3.08 + 0.089 mm. None of the organs showed any kind of gross lesions around the macrocysts embedded in the muscles. Microscopic examination of esophagus and diaphragmatic muscles by squash technique revealed the presence of microcysts arranged horizontally in between the muscle fibers of esophagus where as the pepsin HCl acid digestion of muscle samples of esophagi and diaphragm showed live bradyzoites in gliding motion. Histopathological studies suggested two possible etiologic agents of bovine sarcocystosis namely: S. cruzi, characterized by having elongate and septate cysts and S. hirsuta or S. hominis, characterized by having spherical or rounded cysts with thick radially striated cyst wall. The muscle degeneration with focal or diffused mononuclear cells viz: leukocytic infiltration, eosinophils, lymphocytes and macrophages observed in the tissues under study were attributed to the pathogenic effects of S. cruzi. The ruptured or degenerated state of some of the mature sarcocysts surrounded by eosinophils indicated the advanced age of the cyst. Immunofluorescent antibody technique was standardized in the laboratory for the diagnosis of Sarcocyst infection in bovines. Purified, host cell free bradyzoites collected from macrocysts of Sarcocystis spp, in aliquots of 6-8 applied to glass slides and fixed in chilled acetone over night followed by preservation at -200C worked well with 1:16 dilution of positive and negative control sera and 1:40 dilution of rabbit anti-bovine FITC conjugate. The positive sera did not show any cross reaction with T. gondii RH strain and non specific reactions were absent with negative sera. The serosurveillance of bovine sarcocystosis by laboratory standardized IFAT showed 80.14% (323) of cattle and 78.59% (246) of buffaloes positive for anti sarcocystis antibodies out of 403 cattle and 313 buffalo sera tested, respectively showing an overall prevalence of 79.46% out of 716 animals screened. The antibody titers of 6 randomly selected positive samples from different age groups of <2 years, 2-5 years, 5-10 years and >10 years old bovines ranged from 16-64, 32-256, 32-128 and 16-64 with an average titer of 32 + 2.92, 106.6 + 34, 74.6 + 17 and 34.6 + 9, respectively. The age wise prevalence of sarcocystosis in cattle indicated low rate of infection in the age group below 2 years (60%) and an ascending rate of infection in the age groups of 2-5 years (81.33%), 5-10 years (80.52%) and above 10 years (90.9%). Similarly, the incidence was significantly low in the buffaloes of below 2 years (64%) and high percentage of infection (86.51%) in 5-10 years followed by 78.94% in 2-5 years and 77.27% in above 10 years of age groups. No significant difference of infection was observed between male (81.87%) and female (75.19%) animals as well as between non-descriptive (79.63%) and cross bred (77.58%) animals. Esophageal and diaphragmatic muscle samples collected from 100 animals slaughtered at Chengicherla slaughter house, Hyderabad were subjected to visual examination, squash and pepsin HCl acid muscle digestion techniques which revealed the presence of macrocysts in 16% and 0%, microscopic sarcocysts in 8% and 0% and bradyzoites in 76% and 52% esophageal and diaphragmatic muscles, respectively. The sera collected simultaneously from corresponding animals were screened for anti Sarcocystis antibodies by laboratory standardized IFAT and the results were compared with those of visual examination, squash and pepsin HCl acid muscle digestion techniques. The IFAT was found superior in diagnosing sarcocystosis with positivity of 82%, followed by muscle digestion, gross examination and squash techniques with positive rates of 52%, 16% and 8%, respectively. The present study indicated that the visual and microscopic examination of bovine carcass is by no means a satisfactory diagnostic tool and recommends Immunoflourescent antibody technique for the antemortem diagnosis of animals waiting for slaughter at abattoirs in large scale. Experiments were also undertaken to determine whether Sarcocystis would survive storage at different refrigeration temperatures for a period of 9 days. The number of live and dead bradyzoites in one gram of pepsin HCl acid digested bovine esophageal muscle samples previously stored at room temperature, 40C, 00C, and -200C for a period of 48thhr, 8 days, 24thhr, and 24hr were 2x104 and 4 x104, 1x104 and 1 x104, nil and no bradyzoite, respectively when compared to those stored at 0th hr (10x104 and 0 x104). | en_US |
| dc.identifier.uri | http://krishikosh.egranth.ac.in/handle/1/5810083453 | |
| dc.keywords | BOVINE SARCOCYSTOSIS;IMMUNOFLUORESCENT;ANTIBODY TECHNIQUE;DIAGNOSIS | en_US |
| dc.language.iso | en | en_US |
| dc.pages | 146 | en_US |
| dc.publisher | SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA | en_US |
| dc.sub | Veterinary Parasitology | en_US |
| dc.subject | null | en_US |
| dc.theme | DIAGNOSIS OF BOVINE SARCOCYSTOSIS BY IMMUNOFLUORESCENT ANTIBODY TECHNIQUE | en_US |
| dc.these.type | M.V.Sc. | en_US |
| dc.title | DIAGNOSIS OF BOVINE SARCOCYSTOSIS BY IMMUNOFLUORESCENT ANTIBODY TECHNIQUE | en_US |
| dc.type | Thesis | en_US |