1.
Staphylococcal Coagglutination Test For Rapid Detection Of Foot And Moth Disease Virus
by Baitullah Khan | Dr. Atif Hanif | Prof. Dr. Masood Rabbani | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2010Dissertation note: Foot and mouth disease is a highly contagious, viral disease of cloven hoofed animals and causes high economic losses. Rapid detection of FMD is necessary to control the disease from spreading. Although several reliable tests like ELISA, CFT' and PCR already exit but none of them applicable in field conditions. The aim of this study was to optimize rapid, economical and sensitive test for the detection of FMD. For this purpose rabbits were used to raise immune sera against FMD virus with one uninnoculated control. Immune sera collected from these rabbits at different time interval and presence of antibody was determined by using AGPT. Immune sera were then conjugated with satbalized and inactivated staphylococcus aureus cell using different dilutions. Cell wall of S. aureus contains Protein A which naturally binds with Fc portion of IgG leaving the Fab portion to interact with antigen. In presence of homologues antigen causing agglutination was seen with nacked eyes. Light blue background was found best while observing results.
The coagglutination test was applied on FMD known antigen. Clear agglutination on slide was observed by mixing equal quantity of COAT reagent and its respective antigen. Total 40 vesicular fluid samples from FMD infected animals were tested with COAT, in which 38 yielded positive results and the remaining two yielded negative results. COAT reagents were also tested against PPR virus depicting negative results. COAT was found specific for FMD antigens. This test is quick and generates results within five minutes.
This reagents of CAOT also applied on two fold dilution of vesicular fluid from FMD infected animal and positive result were observed up to 1:32 dilution. This test is sensitive, specific, economical and rapid for detection of FMD. This test was successfully used for detection of FMD in filed.
Availability: Items available for loan: UVAS Library [Call number: 1176,T] (1).
2.
Development And Optimization Of Multiplex Pcr For The Identification Of A, O And Asia 1 Strains Of FMDV In Pakistan
by Muhammad Ikram | Dr. Atif Hanif | Dr. Imran Najeeb | Prof. Dr.
Material type: ; Format:
print
Publisher: 2010Dissertation note: Foot and mouth disease (FMD) is highly infectious disease of cattle, buffalo, sheep and goats. It is caused by genus Aphthovirus of Picomaviradae family. FMDV is RNA virus having seven serotypes A, 0, C, Asia I, SAT1, SAT2 and SAT3.
Foot and mouth disease is endemic in Pakistan and causes high economic losses to livestock industry. So priority is to develop quick and efficient methods for detection of FMDV and to limit the spread of disease outbreak. Although CFT, VNT and ELISA are already being used for the diagnosis of FMDV in Pakistan but these diagnostic techniques are time consuming and their specificity and sensitivity is low. Multiplex PCR for the identification of FMDV is very much sensitive and specific, can be done with in three hours after the receipt of samples.
Present study has been designed to optimize multiplex RT-PCR for rapid detection of FMD virus. RNA was extracted from virus stock obtained from QOL, UVAS Lahore and from field samples. After RNA extraction the samples were subjected to synthesize cDNA by the use of Reverse Transcriptase enzyme. After cDNA synthesis PCR reaction was carried out. The amplified products were resolved on 1.5% Agarose Gel. A multiplex RT-PCR strategy was optimized and developed for the detection of virus serotypes A, 0 and Asia l.
Restulst of this study helped to develop an efficient and economical method for rapid detection of FMD virus and also helpful in differential diagnosis from other vesicular diseases.
Availability: Items available for loan: UVAS Library [Call number: 1189,T] (1).
3.
Preparatuin And Evaluation Of Monospecific Anisera Against Hemagglutinin, Neuraminidase And Matrix Proteins of local Avian Influenza Strains H5 N1, H7 N3, H9 N2, for diagnostics
by Sumaira Ijaz | Dr. Atif Hanif | Dr. Aftab Ahmad Anjum | Prof. Dr.
Material type: ; Format:
print
Publisher: 2011Dissertation note: Avian Infuenza is an economically important disease of poultry worldwide. It has caused losses to poultry industry on larger scale. It is important due to its zoonotic nature. Studies were carried out to raise monospecific antisera against hemagglutinin, neuraminidase and matrix antigens. HA, NA and M proteins of each of the avian influenza strains were separated on Sodium Dodecyl Sulfate Poly Acrylamide Gel Electrophoresis (SDS PAGE). First virus was lysed to release the proteins. Virus was lysed by using 4% Triton X 100, 1mM KCl and 0.01M Tris buffer. Then the sample was dialyzed. Sample was run on gel to purify proteins. The protein bands of appropriate molecular weight were cut and triturated in 1ml of normal saline. Material was centrifuged to remove the gel content.
Each protein was confirmed by the Bradford's Reagent. Each protein was individually mixed with Montanide ISA 50 adjuvant in 1:1 ratio to make the vaccine. Vaccine of each polypeptide of AIV strains was injected in three groups of nine birds each. One group of birds was injected with HA, second group with NA and third group with Matrix proteins of H?, H? and H?. Three groups of birds served as control. The blood samples of all birds were collected before and after inoculating vaccine. The sera of birds before and after inoculating vaccine were checked for antibodies titre against HA antigen by HI test. Antibodies against Matrix antigen were detected by Agar Gel Precipitation Test. Antibodies titre was raised after inoculating polypeptides. In case of sudden outbreaks, antisera may be helpful to control disease.
Availability: Items available for loan: UVAS Library [Call number: 1351,T] (1).
4.
Association Of Myogenic Factor 5 (Myf5) Gene Polymophism With Meat Quantity And Quality Traits In Sahiwal
by Faiza Maqbool | Prof. Dr. Masroor Ellahi Babar | Dr. Abu Saeed Hashmi | Dr. Atif Hanif.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Pakistan has a large population of farm animals, which are consuming for many purposes. Livestock is the major source of money for any country. Livestock is the major machines and factories which convert roughage (grasses and shrubs etc.) into quality food i.e. Meat and milk.
The Myf5 gene is a member of basic helix-loop-helix family of transcription factors which is involved in the regulation of myogenesis. Its main role in muscle growth, development, proliferation, muscle fibers formation and muscle functioning makes it a candidate gene for molecular marker of meat production in livestock.
Myf5 gene in cattle has been mapped to chromosome 5 and has a length of 3236 bp (Gen Bank accession no. NC-007303). It consists of three exons and two introns. Exons have the lengths of 659, 76 and 1245bp. Role of Myf5 gene in muscle development and growth makes it a candidate gene for meat production in farm animals. In this study association of myogenic factor 5 (Myf5) gene polymorphism with meat quality and quantity traits in Sahiwal cattle was checked out.
In this study blood samples were collected from Sahiwal cattle breeds and DNA was extracted from leukocytes. DNA amplification was done by PCR. Then sequencing of amplified gene was done by Genetic Sequencer.
Allele frequencies and genotype frequencies were statistically analyzed by using SNPator software. The relationship between SNP marker genotypes of myogenic factor 5 (Myf5) gene with meat quality and quantity traits was evaluated by using SNPator software. This study will be a helpful tool for marker assisted selection of beef cattle.
Availability: Items available for loan: UVAS Library [Call number: 1561,T] (1).
5.
Biomass Production Of Pasteurella Multocida By Using Biofermentor For Preparation Of Montanoid Based Vaccine
by Noreen Sarwar | Prof. Dr. Khushi Muhammad | Dr. Atif Hanif | Prof. Dr. Muhammad.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Hemorrhagic septicemia is a contagious bacterial disease of large ruminants principally in cattle and buffalo with high morbidity and mortality. The disease is endemic in nature and outbreaks are common during hot, humid and wet season. The acute and fatal nature and brief duration of the disease limit the antimicrobial therapy. In Pakistan, the disease causes heavy economic losses to dairy industry. Vaccination therefore, is an option for controlling the disease. For a quality vaccine, biomass production of P. multocida along with well developed capsule (immunogen) is necessary. The problem associated with the production of a quality vaccine is poor biomass production of P. multocida when grown in ordinary or routine media.
Present study was designed to isolate P. multocida from sick animals and its molecular characterization in the laboratory and study factors (temperature, media composition, pH incubation time and agitation or shaking) affecting its immunogen production and "in process quality control" factors (biological titer, dry mass, adjuvant and storage time) that affect antibody response. Finally, biomass production of the organism using biofermentor and monitoring of the antibody response of buffaloes to inactivated Montanide ISA-70 based P. multocida vaccine.
Each of the field isolates showed grey, viscous, mucoid, translucent and non hemolytic colonies on blood agar. There was no growth on MacConkey's agar. It was Gram negative coccobacilli or thin rods and bipolar when stained with Leishman's stain. The isolates were positive for Catalase, Oxidase, Hydrogen sulphide and Indole production along with nitrate reduction while it was negative for urease production, citrate utilization and gelatin liquefaction. The bacteria fermented glucose, sucrose, mannitol, mannose, but failed to ferment arabinose, maltose, salicin, lactose, dulcito and inositol.
Polymerase chain reaction (PCR) was performed on isolated colonies by using P. multocida specific and HS causing serotype B specific primers. P. multocida specific PCR gave product of 465 bp while HS causing serotype B specific primers amplified a product of approximately 590 bp.
Growth of the bacteria in casein yeast sucrose broth was optimized under different conditions. CSY broth showed dense growth of P. multocida during incubation for 18 hours. A temperature in between 35°C and 40°C showed its optimum growth. Poor growth was observed below 30°C and no growth was detected at 50°C and above. No growth occurred at pH 0.5 and 10.0 but best growth was obtained at pH 7.0 and 8.0. There was positive correlation between shaking in terms of rpm and growth. There was optimum growth at 500 rpm for 24 hours.
Inactivated HS Vaccine was prepared from dense growth in biofermentor on the basis of dry mass and bacterial count. The effect of biomass, adjuvant, storage of the vaccine, priming alone or with boosting on its potency was also studied along with boosting effect of montanoid ISA 70 oil based vaccine. Dry mass 1.7 mg/dose produced protective antibody titer while bacterial count 10-14/ml was sufficient to produce the protective antibody titer. Montanoid ISA 70 based vaccine provided immunity to buffalo calves better than aluminium hydroxide gel and bacterins. Boosting with oil based vaccine can help to keep the animal immunized for whole year. For better results of vaccine, it can be stored at 4oC for six months.
It is concluded that the proposed study improved quality of the vaccine and reduced volume of the vaccine dose, cost of its production and frequency of vaccination.
Availability: Items available for loan: UVAS Library [Call number: 1581,T] (1).
