Clinicopathological Study Of Theileriosis In Naturally Infected Sheep
Material type: Book ; Format:
; Literary form:
Publisher: 2013 Dissertation note: Theleiriosis in sheep is an important infectious disease of small ruminants characterized by General weakness, weight loss, anorexia, elevated body temperature, petechial hemorrhages on conjunctival mucosa, swollen lymph nodes, anemia and cough. Changes in hematological, serum biochemical and histopathological parameters are good indicators for diagnosis of theileriosis. For this purpose a total of 100 blood samples from infected sheep as well as 50 blood samples from healthy sheep were collected on the basis of clinical examination and peripheral blood smears from different farms in and around Lahore. The collected samples were processed at Clinical Pathology Laboratory, Department of Pathology, University of Veterinary & Animal Sciences, Lahore. Hematological parameters were measured by automatic hematology analyzer & serum biochemical parameters were measured by using commercial kits by colorimetric methods using spectrophotometer. Histopathological changes were observed by making slides of tissue samples, and observed under microscope. Molecular confirmation of Theleria sp. was done by using Polymerase Chain Reaction. Finally the data obtained for hematological and serum biochemical parameters were analyzed by using Student's t test.
Previous studies on serum biochemical parameters and hematological parameters were mostly done in large ruminants. But there is little investigation available on the biochemical and hematological alterations of theileriosis in small ruminants. In this study blood samples taken from sheep were examined under microscope by making thin blood smears using Giemsa stain. Theileria piroplasms were seen in RBCs at 100x. Blood samples were also processed for PCR test. In all Theileria ovis positive samples 520-bp fragment was generated on gel by using primers TSsr 170F and TSsr 670R for sheep. All samples positive on microscopy were also positive by PCR. This study provides the base line data for molecular diagnosis of Theileria ovis in sheep in Pakistan. Hematological and biochemical parameters were evaluated in the sheep naturally infected with theileriosis while healthy sheep were selected as control. Hematological findings revealed a significant decrease (P<0.05) in RBC count, WBC count, Hb Conc. and PCV in infected animals as compared with healthy animals. Serum biochemical findings also revealed alterations in activities of enzymes and plasma proteins. A significant decrease in total proteins, albumin, glucose and creatinin and triglycerides was observed while significant increase (P<0.05) in ALT, AST, bilirubin was observed in affected animals as compared with healthy animals. A non significant increase in urea and cholesterol concentration was also observed in infected animals as compared to healthy animals. Different tissues were examined to study gross and histopathological changes. Liver was large, pale and friable. Prescapular lymph nodes were swollen and spleenomegaly was also seen. Kidney showed hemorrhagic spots and some were mottled. Histological examination of the lymph nodes revealed edema, widened intercellular spaces, parenchyma degeneration, massive lympholysis, widened sinuses at some areas and macrophages. Splenic nodules were sparse, diminished in size and lymphocyte depletion in follicles was prominent.In liver, mild infiltration of leukocytes around blood vessels, vacuolation in hepatocytes, increase in sinusoidal space and atrophy of hepatic cord was seen and hepatic cord was broken. In kidney, degeneration of tubular epithelium cells was seen. This study concluded that theileriosis in sheep was associated with some alterations in blood parameters and histopathological changes which could be useful in the diagnosis of ovine theileriosis.
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Pathobiological Investigations Of Peste Des Petits Ruminants (Ppr) Virus With Reference To Antiviral Activity Of Nigella Sativa (Black Seed)
Material type: Book ; Literary form:
Publisher: 2017 Dissertation note: Peste des Petits Ruminants (PPR) is a highly contagious, infectious, acute or sub-acute transboundary viral disease of domestic and wild small ruminants. It is an economically important viral disease of sheep and goats causing varying degree of morbidity and mortality in susceptible animals which may be as high as 100 and 90 per cent, respectively. PPR is responsible for serious socioeconomic problems. There is no data available regarding pathogenesis and field virus characterization to compare it with vaccinal strain for any difference. Nigella sativa(Black Seed) has antiviral activity against many viruses. Therefore present studywas undertaken to investigate the antiviral effect of Black Seed in vivo and in vitro against PPR virus. Further more time course detection of virus is still needed to be studied.
Nigella sativa (Black seed) has antiviral activity against PPR virus.
Pathogenesis can better be studied through histopathology, necropcy findings and morphometric changes.
A total of 250 clinically positive samples suspected for PPR virus were included in the study. Samples were consisted of nasal, ocular and anal swabs; whole blood in EDTA were collected from suspected animals. In case of mortality morbid material included lungs, liver, spleen and mysenteric lymph nodes were included in the study. Samples were subjected to immune capture Elisa for detection of viral antigen in suspected samples. Samples which found positive foe IC – Elisa were then subjected to RT-PCR for confirmation of virus. After confirmation of virus through IC – Elisa and RT-PCR the positive samples were subjected to virus isolation on vero cell. After isolation of virus, the TCID 50 of the virus was calculated for preparation of inoculum for further use. In this experiment mesenteric lymph nodes and spleen found to be major organ for isolation of PPRV.RT-PCR found to be most reliable and confirmatory diagnostic test for PPRV. Field Virus adaptation on vero cells found to be difficult to optimize.
In this experiment antiviral activity of black seed was checked on vero cells infected with PPRV. Three extracts of N. Sativa were prepared to check the in vitro antiviral activity of black seed. In this study poly saccharides extracted from black seed found to be more effective against PPRV. Adaptation of field virus was done on Vero cell line. Antiviral activity of Black Seed extract was determined in vitro on Vero cell on bases of CPE (Cytopathic effect). The ethanolic and aqueous extract were found to be more toxic to consistency of monolayer of vero cells. The TCID50 of virus was calculated after treating cells with different extracts. In this study poly saccharides extract exhibit lower TCID50‘s as compared to ethanolic and aqueous extract which showed higher TCID50’s.So less cytopethic effect was observed in vero cells treated with black seed extracts. Antiviral activity was determined on base of CPE.
Pathogenesis of virus in natural host was studied through time course detection of virus in body secretions, blood, organs. Histopathological changes were studied.20 goats were procured from market divided into four groups (n=5) A,B,C and D. In animals of group A prophylactic effect of N.Sativa was studied. In group B complete pathogenesis of PPR virus was studied without any prophylactic or therapeutic measure. In group C therapeutic effect of N. Sativa was studied after onset of clinical picture of disease. At the end of this experiment, clinical picture, gross pathology, histopathology, and morphometric changes revealed that N. Sativa has noticeable prophylactic effect on PPR infected goats. It can be used as a therapeutic agent in PPR infected goats but it can’t control pathological effect of virus after onset of infection.
Data collected were statistically analyzed by using Microsoft Excel (Microsoft Excel, 2007) and SPSS (for Windows, Version 16.0). The data were put the descriptive analysis and Chi square test was employed to test the significance and test of hypotheses
It was concluded that Black Seed therapy possessed marvelous prophylective effect against PPR virus and RT-PCR was the most efficient methodology to confirm the virus.
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