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1. Comparative Efficacy Of Passive And Active Immunization During Newcastle Disease (Nd) Outbreak In Broilers

by Mushtaq Ahmad Gondal | Prof.Dr.Irshad Hussain | Prof | Prof.Dr.Khushi Muhammad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2007Dissertation note: Newcastle disease is an economically important disease of poultry resulting in huge economic losses every year to the poultry farmers in Pakistan. To compare passive immunization and active immunization during outbreak of Newcastle disease a total of 140 chicks at 16th day of age were divided into seven groups (A, B, C, D, E, F and G) containing 20 birds in each. The level of maternal antibody in chicks, was determined by haemagglutination inhibition titres which revealed that it was the highest at one day and decreased with increasing age. Newcastle disease virus gifted from Dr. Shafqat Fatima Rehmani, Director, Poultry Vaccine Center, Karachi, Sindh was pathotyped by using MDT and ICPI. The Embryo Lethal Dose5o was calculated to be 1083h/0. imi and was found highly pathogenic. Infection was induced in birds through administrating 100 ELD5O1O631/0.lml of Velogenic Newcastle disease virus. Birds of group A, this group seved as a negative control. In group B, this group acted as a positive control. Infection was given by using 0.1 ml of 100 ELD50 of Velogenic Newcastle disease virus intranasally at 26 days of age. In group C, at 16th days of age, all birds in this group were vaccinated with Newcastle disease virus vaccine. At 26th day of age, this group was exposed to infection as mentioned above. In group D, at 24th days of age, all birds in this group were vaccinated with Newcastle disease virus vaccine. At 26th day of age, infection was given by using 0.1 ml of 100 ELD50 of VNDV intranasally into individual bird. In group E, infection and vaccination were given simultaneously at 31 day of age. In group F, In this group, infection was given by using 0.lml of 100 ELD50 of VNDV intranasally, at 26 days of age. When Newcastle disease symptoms were noticed, birds were vaccinated with lentogenic strain of Newcastle disease virus vaccine (Lasota, TAD- Germany) by using 0.5mllbird orally. In group G, infection was given by using O.lml of 100 ELD50 of VND.V intranasally, at 26 days of age. When Newcastle disease symptoms were induced, birds were treated with 64 units of anti-NDV-haemagglutination inhibition yolk antibodies. Use of Lasota vaccine and preformed antibodies in yolk help in decreasing economical losses due to outbreak of Newcastle disease in poultry. Availability: Items available for loan: UVAS Library [Call number: 0995,T] (1).

2. Electrophoretic Profile Of Normal And Hydropericadium Virus-Infected Liver

by Shahid Khan | Prof.Dr.Irshad Hussain | Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2007Dissertation note: Hydropericardium syndrome primarily affects the broilers between the ages of 2-7 weeks. 1he vaccine prepared from infected liver extract treated with formaldehyde is being used to protect the broilers from the disease. The current study was carried out to check the presence of immunogenic proteins in commercially available HPS vaccines Hyper immune serum was raised in 60 (3 weeks old) broiler birds using different commercially available HPS vaccine. The sera were subjected to AGPT for screening out HPS positive serum samples. The HPS infected livers were homogenized and loaded in alreadv prepared agar gel plates. The plates were incubated for 48 hours at room temperature in humid plastic box. Only 5 liver samples were found positive .The HPS autogenous vaccines,HPS positive liver samples normal liver samples were subjected to homogenization, sonication, chloroform treatment, ammonium sulphate precipitation, mixed with equal volume denaturing buffer and then boiled for 3 minutes to extract total proteins of Sample. . Large variations in the protein loading of samples in adjoining lanes lead to distortion. Spectrophotometer determination of protein concentration assay were used to quantitate the known protein samples (bovine serum albumen is commonly used standard or this method) to standardize protein concentration. A28o were used to determine protein concentration and a calibration curve was created by plotting and performing regression analysis of A28o versus concentration of the standards and absorbance of the sample were used to determine the concentration from the calibration curve by using the formula. (concentration =Standard Factor* Dilution Factor* Optical Density. Total protein analysis was carried out by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting technique. A typical gel of 9% acrylamide composition was used to nicely separate polypeptides of samples to analyze the entire profile of a fraction that contains heavy and light polypeptides. Best results were obtained when 30 pi of a 20-30 mg/mi final concentrations of denatured protein sample were loaded per sample well .Each sample was repeated twice on gel. The gel was cutted at the centre vertically. 0.1% Coomassie Blue dye in 50% methanol, 10% glacial acetic acid were used to stain half of the gel for detecting protein while half of the gel was subjected to western blotting. Relative molecular weight (MW) of each protein fraction was determined by plotting a standard curve. The western blot analysis of proteins of hydropericardium syndrome virus infected liver and HPS autogenous vaccine, separated on 9% gel showed one immunogenic protein, molecular weight 15-20 kDa. However, further studies are needed to establish its immunogenic nature and feasibility for its use as vaccine. Availability: Items available for loan: UVAS Library [Call number: 1005,T] (1).

3. Comparative Neutralizing Effect Of Mycotoxin Binders And Its Role In Improving Humoral Immune Responses

by Hazrat Nabi | Prof.Dr.Irshad Hussain | Dr.Aftab Ahmad Anjum | Prof | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2008Dissertation note: µPoultry industry in Pakistan provides inexpensive meat for almost everybody, rich or poor. Poultry farming which once was a blooming business now faces many challenges. Although, infectious agents constitute a major challenge for this industry, mycotoxins are also considered a threat for poultry business worldwide. Mycotoxins, especially aflatoxins are very common in the feed stuff used in the assembly of poultry feeds. The ill-effects associated with aflatoxins are well documented. There are toxin binders marketed by various companies who trumpet various beneficial effects of using these products in poultry. The present project was undertaken to corroborate their claims and also to see whether the use of such products may not result in ill effects that might impact negatively on various production parameters of poultry. Two commercial products (Mycotox® and Mycofix® Plus 3.0) were used to study their contribution on the immune response against ND, feed consumption, weight gain, feed conversion ratio and "lymphoid organ body weight ratio" in broiler chickens. Asp ergillus parasiticus was used for the production of the aflatoxin which was fed to various groups of birds.One hindered and eighteen day-old broiler were randomly divided into 6 groups viz. A, B, C, D,E and F each comprising 18 chicks. The chicks in each group were further randomly sub-divided into 3 replicates comprising of 6 chicks in each. Group A= Only Feed (Negative Control for AF),Group B= AF @150 µg/Kg of feed. (Positive Control for AF), Group C Mycotox® @ 1 gum/Kg 'feed (Positive Control for Mycotox®), Group D= Mycofix® Plus 3.0 @ 2.5gm/Kg of feed. (+ Control for Mycofix® Plus 3. Q), Group E= AF 1 50 µg/Kg and Mycotox® @ 1 gm/Kg of feed (Experimental Group for Mycotox®), Group F AF @ 150 µg/Kg and Mycofix® Plus 3.0 @2.5gm/kg of Feed (Experimental Group for Mycofix® Plus 3.0). The rice powdered material containing AF was mixed according to the calculation to get desirable level of AF (150 µg/kg) in the feed. The birds were vaccinated against Newcastle Disease at the age of 6th days (Intra-ocular route) and then at 24th day (drinking water) and were d libitum. Effects of adding mycotoxin binders to feed containing 1 50ppb aflatoxin were in broiler chicks from 14 to 42 day of age. Compared to the B (positive control for aflatoxin) fed aflatoxin alone significantly reduced geometric mean titres, feed consumption, body weight gain and feed conversion ratio. However, no differences in GMT, body weight gain and feed conversion ratio were found between the chicks fed mycotoxin binders C (positive control for Mycotox®) and D (positive control for Mycofix® Plus 3.0) or mycotoxin binders plus aflatoxin treatment groups E (Experimental Group for Mycotox®), F (Experimental Group for !vlycofix® Plus 3.0) and the control group A (Negative control for Aflatoxin), indicating apparent protection against the deleterious effects caused by aflatoxins. Treatment related changes in organ body weight ratio of thymus, spleen and Bursa of fabricius were also observed. Most of the parameters measured for the birds fed mycotoxin binders did not alter. The addition of mycotoxin binders to aflatoxins contaminated feed diminished the adverse effects of aflatoxins on antibody titres against Newcastle and most relative organ weights. These findings suggested that mycotoxin binders can effectively reduce the toxicity of aflatoxin in broiler chicks and mycotoxin binders can be potential ameliorator against aflatoxicosis in broiler chicks. Availability: Items available for loan: UVAS Library [Call number: 1020,T] (1).

4. Electrophoretic Profile Of Cellular Proteins Of Staphylococcus Aureus From Mastitic Cattle

by Muhammad Zubair Munir | Prof.Dr.Irshad Hussain | Prof.Dr.Masood Rabbani | Prof.Dr.Zafar | Faculty of Veterinary Sciences.

Material type: book Book; Format: print Publisher: 2008Dissertation note: Decline in milk production has. mostly been attributed to infectious agents that generally result in swelling of udder, changes in the constitutions of milk and finally induration of the udder which causing huge economic losses in livestock production. Mastitis is most important diseases of all the lactating animals. The most common Staphylococcus aureus which causes mastitis in cattle. Given the range of ill-effects of Staphylococcus aureus causing mammary infection in cows and buffaloes in Pakistan there is strong need for detection of strain involved. Likewise identification of clones with extensive geographic distribution wall provide insight into strain virulence and pathogenesis and also requiring public health intervention such as vaccination and antimicrobial restriction aimed at reducing the spread of the pathogen. In the present project the relatedness by elcctrophoretic profile of Staphylococcus aureus isolates involved in mastitis at dairy farns were studied, to compare the isolates of Staphylococcus aureus from different dairy farms. The samples from clinically/ subclinically affected at farms were collected, for isolation of Staphylococcus aureus, these samples were streaked onto Staph- 110 agar media plates and Staphylococcus aureus were be confirmed through biochemical and sugar fermentation tests. These conformed Staphylococcus aureus Isolates were subjected to sodium dodecyl sulphate polycrylaniide gel electrophoresis. (SDS-PAGE) for electrophorctic protein profiling. The banding pattern of various isolates as against known molecular weight markers were recorded for interpretation of results. It was concluded that protein profile is one of several methods for determining the relatedness or unrelatedness of bacterial strains. As immunization has been often attempted in efforts to control the mastitis, the diversity of Staphylococcus aureus strains and difficulties in vaccine development instruct that use of one or likely several strains in a herd or even in herds located in the same geographical region are recommended. Decision on representative strains can be made on basis of whole cell proteins, since such proteins of Staphylococcus aureus strain has already been shown, the isolates could be characterized by their total protein profiles and use of one or likely several strains of Staphylococcus aureus that are antigenically representative of the majority of the causative strains will overcome vaccine development difficulties. Availability: Items available for loan: UVAS Library [Call number: 1047,T] (1).



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