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1. Production, Purification And Evaluation Of Anti Tetanus Serum

by Mian Muhammad Khubaib Sattar | Prof. Dr. Tahir Yaqub | Dr | Mr. Muhammad Zubair Shabbir.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2012Dissertation note: To produce anti-tetanus serum, ten female sheep of about 3 years of age are immunized with commercially available tetanus toxoid vaccine (Imatet™, Amson vaccines and Pharma) for eighteen weeks to these experimental animals with two weeks interval. Serum samples of all the sheep were also collected fortnightly and stored at -20 ºC. ELISA was performed to determine the antibody (IgG) titer of all the test samples. Out of 90 samples, 20 samples showed tetanus antibody (IgG) titer of 100.8 I.U. or more, while 8 samples presented tetanus antibody (IgG) titer of 160.9 IU or more. Out of these 8 samples, 3 samples had tetanus antibody (IgG) titer of 190.9 I.U. or more. Maximum tetanus antibody (IgG) titer was 195.4 I.U. at day 120. Three samples exhibiting maximum antibody titer (190.9 I.U., 195 I.U. and 195.4 I.U.) were mixed in equal quantities for purification of Immunoglobulins (IgG). A volume of 15 mL of aggregated serum samples was mixed with 15 ml of saturated ammonium sulfate having final concentration of 45 % in the mixture which is continuously stirred at room temperature for 1 hour. Mixture was centrifuged at 10000 rpm for 30 minutes in refrigerated centrifuge machine and dialyzed against 10 changes of PBS at 4 ºC. Desalting is checked with BaCl2 solution. The purified tetanus immunoglobulins (IgG) were treated with papain to produce Fab Fragments. Then the protein content of the purified tetanus immunoglobulins and Fab fragments was estimated with Bradford protein Assay. BSA standard curve was used to produce a regression equation [Y (OD Value) = 0.218 + 0.033 X (Protein Concentration)] which was used for calculation of the protein contents of the samples. The purified tetanus immunoglobulins and Fab fragments were tested for purity with SDS-PAGE analysis. Then in vivo toxin neutralization test was performed in mice to check the tetanus toxin neutralization ability of the sera produced. Availability: Items available for loan: UVAS Library [Call number: 1420,T] (1).

2. Seroprevalence And Risk Factor Analysis Of Bluetongue Virus In Lahore And Faisalabad Districts Of Punjab Province, Pakistan

by Syeda Marriam Maqbool (2014-VA-522) | Dr. Muhammad Zubair Shabbir | Dr. Ali Ahmad Sheikh | Dr. Maryam Javed.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Domestic animals play an important role in the rural and agricultural economies of developing countries. Therefore, animal diseases pose a threat to country’s economy, animal welfare, the environmental and public health. One of the important animal diseases is Bluetongue, listed as notifiable disease by OIE. Causative agent is Bluetongue virus (BTV) an arbovirus that belongs to genus Orbivirus with in family Reoviridae. The main route of transmission is through the bite of Culicoides biting midges. Disease is enzootic and widely distributed in areas where susceptible animals and vector species are prevalent. It has been reported worldwide including the neighboring countries of Pakistan. BTV is also considered an endemic in Pakistan but little information is available on its epidemiology in this area. Serological tests can detect antibodies produced against infection and helpful to analyze the prevalence of a pathogen in circumstances when there lacks vaccination practice to ruminants in a given geographical area. Competitive ELISA was used to identify antibodies to BTV in the sera samples of animals in Faisalabad and Lahore districts. Blood samples were collected from randomly selected villages of both districts and processed for serum separation by using gel/clot activator tubes. Separated serum was analyzed by competitive ELISA. Further, statistical analysis was done by OpenEpi to check the association between BTV seroprevalence and potential risk factors. Later the BTV prevalence has been mapped in relation to different villages of both districts. Results of present study revealed that Bluetongue virus is prevalent in Faisalabad and Lahore districts with high seropositivity observed for Faisalabad district. Antibodies to BTV were detected in all studied animals irrespective of their age, sex, parity and breed. Risk factor analysis is implicating the association of BTV seroprevalence with breed, sex and age for sheep, SUMMARY 44 cattle and buffalo respectively. Further studies should be conducted to expand the geographical area for the assessment of Bluetongue prevalence and to explore the genetic diversity of Bluetongue virus. Availability: Items available for loan: UVAS Library [Call number: 2497-T] (1).



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