1.
Comparative Immunopathological And Immunosuppressive Effects Of Three Different Gumboro Vacine Strains
by Rana Muhammad Ayyub | Dr. Asim Aslam | Dr. Shakil Akhtar Khan | Faculty of Veterinary Sciences.
Material type: ; Format:
print
Publisher: 2002Dissertation note: This project was planned to know the comparative immunosuppressive effects of three different Gumboro live vaccine strains on ND vaccination and to choose the least immunosuppressive out of these three vaccine strains.
A total of 100 chicks were divided into 4 groups (A, B, C and D), 25 birds in each group. All the birds of all the groups were vaccinated against Newcastle disease on 5th and 21st day of age. Specific IBD vaccine was given to specific groups at l4t1 day and 28th day of age. Group A was given intermediate plus strain vaccine (228 E strain), group B was given intermediate strain vaccine (Strain D78) and group C was given another intermediate strain vaccine (Bursine-2 strain), on both days of IBD vaccination, as per schedule. While group D was unvaccinated control.
Immune organs including bursa, thymus and spleen were examined for their gross and microscopic pathological changes, before and after IBD vaccination. For this purpose, these organs were collected at 13th j7th and 31st days of age. At j3th day, gross and histopathological examination showed no lesions in any case of any group (as so far no IBD vaccine was given ). At 17th and 31st days, gross and histopathological examination revealed more severe lesions in group A, moderate lesions in group B, mild lesions in group C and no lesion in immune organs of group D.
Haemagglutination inhibition (HI) test was performed, to study the comparative humoral immunosuppressive effects of these IBD vaccines on antibody titre against ND vaccination, on birds of each group. Statistical analysis showed that 228 E vaccine strain (group A) while, D78 vaccine strain (group B) was less immunosuppressive but, on the other hand Bursine-2 vaccine strain (group C) was least immunosuppressive. No humoral immunosuppression was observed in unvaccinated control group D.
This study suggests the use of Bursine2 strain of IBD vaccine, in a flock at risk of ND infection, as it has least immunosuppressive effect against ND vaccination. Therefore, still the need of the hour is to choose the most proper IBD vaccine strain, among all the available strains in the market, which can provide excellent protection against IBD infection as well as has least immunosuppressive effects on ND vaccination.
Availability: Items available for loan: UVAS Library [Call number: 0775,T] (1).
2.
Immuo-Pathological Response Of Pigeons To Challenge Infection Of Newcastle Disease Virus (Ndv)
by Yasir Amin | Dr. Asim Aslam | Prof. Dr. Muhammad Younus Rana.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2011Dissertation note: This study trial was designed to evaluate and compare the efficacy of two locally available live Newcastle disease vaccines (Medivac ND LaSota and VRI, Mukteshwar ND vaccine), also to compare two routes (Oral and Ocular) of vaccine administration in term of antibody titre and assessment of protection against field (chicken) isolated virulent Newcastle virus challenge (NDV) in pigeons. Study of clinical signs, gross and histopathological lesion in different organ of non-vaccinated and challenged birds was also the part of our present study. For this purpose one hundred and twenty pigeons were purchased from the local market and screened for Newcastle disease antibodies using Hemagglutination inhibition test. Healthy pigeons were randomly divided into six groups i.e. A, B, C, D, E and F, comprising 20 birds each. Group E and F were kept as positive and negative Control respectively. Group A and C were vaccinated with Medivac ND LaSota vaccine at day 7th and 21st of experiment through oral and ocular route. Similarly Group B and D were immunized with VRI (Mukteshwar) ND vaccine through oral and ocular route respectively. At 28th day of experiment all the groups except group F were challenged with velogenic field isolate of NDV at a dose rate of 0.1 ml through ocular route. Serum samples were collected at day 14, 21, 28, 35 and 42 of experiment for the determination of antibody titre. Post-infection clinical signs in control positive group were i.e. anorexia, dullness, depression, decreased feed intake, discharge from mouth, greenish diarrhea, nervous manifestations, leg and wing paralysis. Gross lesions on different organs were hemorrhages in trachea, proventriculus, spleenomegaly and greenish intestinal contents. Medivac ND LaSota vaccine produced higher immune response in term of antibody titre as compared to VRI (Mukteshwar) ND vaccine. It was also observed that ocular route irrespective to vaccine type produced significantly (P<0.05) higher immune response than oral route. Vaccine strains used in this study efficiently induced immune response through ocular route, suggesting that implementation of this vaccination programs in future may prevent ND outbreaks in pigeons, especially in racing pigeons, and may prevent NDV spread to other avian species, mainly poultry.
Availability: Items available for loan: UVAS Library [Call number: 1228,T] (1).
3.
Clinicopathological Study Of Theileriosis In Naturally Infected Sheep
by Muhammad Waseem Akhter | Dr. Asim Aslam | Dr. Muti ur Rehman Khan | Prof. Dr. Habib.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2013Dissertation 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.
Availability: Items available for loan: UVAS Library [Call number: 1642,T] (1).
4.
Immuno-Modulatory Effects Of Beta-Glucans And Nucleotides Based Commercial Products In Broiler Chicken Experimentally Infected With Velogenic Newcastle Disease Virus
by Hafiza Zain ul Fatima (2008-VA-232) | Prof. Dr. Asim Aslam | Dr. Raheela Akhter | Dr. Aamir Ghafoor.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Immune-potentiation effect of commercially available products containing beta-glucans (betaplex and catalyzer) and nucleotides (electroimmune and avimmune) in the broilers was evaluated. A total of 120 broiler chicks were reared under standard husbandry conditions. The birds were vaccinated by live NDV vaccine at 5, 14 days and killed Newcastle disease virus (NDV) vaccine at 7 days of age. Additionally, live infectious bronchitis and infectious bursal disease vaccines were also given through water at 5 and 10 days of age, respectively. The birds were randomly divided into six groups each comprising 20 birds. Four groups (treatment groups) were offered four commercial immune-booster products at 7, 14 and 21 days of age for 72 hours. Fifth group (untreated vaccinated) was not offered any drug while sixth group (untreated and unvaccinated) was not given any drug and also remained unvaccinated against NDV. Anti NDV antibodies were measured at 7, 14, 21 and 28 days of age and heterophil/lymphocyte (H/L) ratio was measured at 14, 21 and 28 days. Weight of immune organs and performance of birds was also measured from each group. Finally, 15 birds from each group were inoculated with virulent NDV through intranasal route at 28 days of age to assess the protection against challenge infection.
Mean GMT anti NDV antibody titers in vaccinated groups were 3.65 (±0.59) at one week of age which were raised to the levels of 5.85 (±0.88), 5.50 (±1.00), 6.10 (±0.64), 5.90 (±0.91) and 5.45 (±1.05) in the betaplex, catalyzer, electroimmune, avimmune, and untreated control groups, respectively. Maximum antibody titer was achieved in electroimmune treated group while minimum in the catalyser treated ones. In non-vaccinated control group the antibody titers were minimum in the beginning that reduced to negligible level at four weeks of age. The H:L ratio of
Summary
48
all treatment groups did not significantly vary at 14 days of age while at 21 and 28 days of age, the birds receiving electroimmune have significantly low H:L ratio in comparison to other treatment groups. Weight of liver and spleen in nucleotide treated groups was significantly higher than other treatment and control groups while there was no significant difference in the weight of bursa of fabricious in all treatment groups. Relative weight of all immune organs to the live body weight did not significantly vary in all treatment and control groups. Performance of the birds as a measure of feed conversion ratio (FCR) was highest in electroimmune treated birds. The challenge protection data shows that minimum mortality of 40 % (6/15) was observed in nucleotides treated groups while betaplex and catalyzer treatment groups have 47 % (7/15) and 53 % (8/15) mortality ratios in comparison to 60 % (9/15) mortality in untreated birds. Results of present study suggest that nucleotide or beta glucans supplementation in the diet have beneficial effects to enhance the humoral immune response and better bird performance in broilers. Use of these products not on delay the onset of clinical disease but also helpful in providing better protection against challenge to velogenic NDV. Availability: Items available for loan: UVAS Library [Call number: 2516-T] (1).
5.
Amelioration Of Pathological Changes Due To Infectious Bursal Disease By The Administration Of Mentofin And Asi-Mirus In Broiler Chicken
by Muhammad Umair Shah (2011-VA-15) | Prof. Dr. Asim Aslam | Dr. Ghulam Mustafa | Dr. Ali Ahmed Sheikh.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Poultry industry is the second largest industry in Pakistan but despite of its rapid growth rate it is facing huge economic losses due to many infectious diseases. Infectious Bursal Disease is one of them. Huge economic losses in case of infectious bursal disease are due to immunosuppression and high mortality.
In Pakistan, commercially available vaccines are abruptly used to control different viral diseases but unfortunately failure of these products occur from time to time. Hence, current study was designed to determine the immunostimulatory effect of two commercially available products (Mentofin and ASI-MIRUS) against IBD vaccine.
A total 300 broiler chicks were taken, divided into six groups each having 50 birds and were replicated under controlled conditions. A, B and D groups were vaccinated with the IBD live virus vaccine. B and C groups were treated with Mentofin. D and E groups were treated with ASI-MIRUS while F group served negative control. To detect antibody titer against IBDV at every week (0-42 days of age), a commercial ELISA kit, IDEXX Flock Chek standard (IDEXX Corporation, Westbrook, ME, USA) was used. In order to analyze gross and microscopic changes in bursa, postmortem examination and histopathology of bursa was done.
The volatile oils in Mentofin and ASI-MIRUS have effective immunomodulatory effects on humoral immune response in broiler chicks. Eucalyptus and peppermint oils increase bursa to body weight (B/BW) ratio as compared to untreated birds. Results of present study indicated the highest antibody titer in group D supplemented with ASI-MIRUS and vaccinated as compared to group B supplemented with Mentofin and vaccinated. Significantly high bursa to body weight ratio also observed in vaccinated group D (ASI-MIRUS treated) comparing with other
CHAPTER 6
SUMMARY
Summary
35
vaccinated groups A and B. In Group B (Mentofin treated), bursal samples showed necrosis at medullary region of bursal follicle. Group D (ASI-MIRUS treated) showed the active follicle consist of lymphoid cells and shown no obvious histopathological lesion. So present study showed that ASI-MIRUS is reduced the severity of IBDV which has more beneficial effect on immune response against IBD vaccinated Broiler Chicken as compared to Mentofin. Availability: Items available for loan: UVAS Library [Call number: 2557-T] (1).
6.
Pathogenesis Of Aflatoxin B1 In Quails Under Experimental Conditions And Detoxification By Biological And Chemical Means
by Sakhra Mahmood (2005-VA-251) | Prof. Dr. Muhammad Younus Rana | Prof. Dr. Asim Aslam | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Secondary metabolites of certain fungi produce toxins under favorable conditions especially while growing on different food grains. Mycotoxins are among major threats to growing poultry industry and human beings. Aflatoxins are closely related, biologically active fungal metabolites and commonly produced by Aspergillus species.
A research was carried out to evaluate the ability of Aspergillus flavus for Aflatoxin B1 production using rice, wheat and maize as substrates. Lethal effects on growth performance parameters, hematological and histopathological of graded doses of aflatoxin B1 in quails under experimental conditions were observed. Effect of Aflatoxin B1 on humoral immune response to Newcastle Disease virus vaccine in quails were determined. Biological detoxification of Aflatoxin B1 by Saccharomyces servisiae was evaluated in quails. Comparative evaluations of different commercially available toxin binders were checked. All these experiments were carried out till the six weeks (42 days).
Aspergillus flavus was identified on the basis of macroscopic and microscopic characteristics. Rice, wheat and maize grains was used as substrate to check the level of Aflatoxin B1 produced by inoculating an aqueous suspension of 106 spores/ml. Aflatoxin B1 checked by Thin Layer Chromatography (TLC) and quantified by High Performance Liquid Chromatography (HPLC).
Quails were reared under standard management conditions in five groups (A, B, C, D and E) having sixty each. Each group was further divided in two independent units. Diets offered to groups were control (without toxins), 0.25, 0.50, 1 and 2 mg Aflatoxin B1/kg feed. One unit of
SUMMARY
187
each group was vaccinated with Newcastle Disease Virus (NDV) vaccine while other was not and studied the lethal effects on growth performance, blood parameters, immune response and histopathology of vital organs. At the end of the experiment, it was found that the deleterious effects of Aflatoxin B1 were dose and duration dependent. As the level of the toxin was increased, the lethal effects were prominent. The growth performance parameters including gain in body weight, feed intake and feed conversion ratio was adversely affected at high doses. The body weight gain was significantly reduced in Aflatoxin B1 treated groups as compared to control group. Similarly feed intake and feed conversion ratio were significantly different from the control group. The hematological studies exhibited that aflatoxin B1 significantly reduced the hemoglobin, packed cell volume and total leukocyte count whereas the erythrocyte sedimentation rate was significantly increased as compared to control group. The immune response against NDV vaccine was adversely effected in Aflatoxin B1 treated groups and values of Antibody titer in AFB1 were significantly low as compared to group A( control) In the second experiment, Saccharomyces cervisae (SC) dried powder was mixed in basal quail diet having 0.5mg Aflatoxin B1 for all experimental groups and control was without toxins. SC was added at levels of 0.5 gm, 1.0 gm and 2.0 gm /kg of feed. It was recorded that Saccharomyces cervisae (yeast) have the potential to remove the deleterious effects of Aflatoxin B1. Yeast effectively detoxified the Aflatoxin B1. The results recorded of growth performance and other parameters were non-significantly different from the control group. Chemical detoxification of Aflatoxin B1 was evaluated in quails using commercially available toxin binders. Toxin binders used were activated charcoal, kaoline, Myco AD and selenium plus vitamin E and mixed in basal quail diet having 0.5mg Aflatoxin B1 for all experimental groups and control was without toxins. The Myco AD and selenium plus vitamin E showed the highest detoxification potential as compared
SUMMARY
188
to other chemical toxin binders. Groups E and F showed the results of growth performance, hematological, immune response and histopathological were non-significantly different from the control group (A). Kaolin was moderately detoxifying the toxin.
Presence of aflatoxin B1 in soft tissues was checked by TLC and quantified using HPLC. The liver exhibited the residues of Aflatoxin B1 at high doses of toxin. Group D and E rearing on feeds having 1mg AFB1 /Kg feed and 2mg AFB1 /Kg feed of toxin showed the residues of AFB1 in liver and kidney.
Statistical means for growth performance parameters, hematological, immune response and histopathological scores in each subunit of quails were analyzed by applying one way ANOVA and Duncans‟s Multiple Range (DMR) test at 95% probability. Aflatoxin B1 is lethal and lowers the performance of birds. The lethal effects can be detoxified by biological and chemical means to lower the economic losses to poultry industry. It can be concluded that biological detoxification is preferably better as compared to chemical detoxification. Availability: Items available for loan: UVAS Library [Call number: 2670-T] (1).
7.
Pathobiological Investigations Of Peste Des Petits Ruminants (Ppr) Virus With Reference To Antiviral Activity Of Nigella Sativa (Black Seed)
by Kiran Aqil (2008-VA-456) | Dr. Muti Ur Rehman Khan | Prof. Dr. Asim Aslam | Dr. Aqeel Javeed.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation 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.
SUMMARY
130
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. Availability: Items available for loan: UVAS Library [Call number: 2890-T] (1).
