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1. Comparitive Immunogenic Evaluation Of Purified And Whole Culture Vaccine Of Pasteurella Multocida B2 Of Bovine Origin

by Anika Khalid (2006-VA-357) | Dr. Imran Altaf | Dr. Jawad Nazir | Dr. Wasim Shehzad.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Pasteurella multocida is responsible for causing Heamorrhagic Septiceimia which is a fatal and acute disease mainly associated with bovines. Pasteurella multocida consists of two important antigenic structures i-e Outer membrane protein(OMP) and Lipopolysaccharide(LPS) that forms capsule.These two structures played vital role in pathogenicity as well as in the immunogenicity. In our present project we study the role of LPS,OMP alone and in combination as an immunogens.We compared the immunogenic behaviour of whole culture vaccine(consists of LPS and OMP) purified vaccine(consists of OMP),LPS vaccine (consists of LPS only) by IHA and CFT (using OMP and LPS both as an Ag separately) and the overall results showed that whole culture HS Alum precipitated vaccine produced better antiboby titer against both outer membrane proteins (OMP) and lipopolysacharrides (LPS) when checked against respective antigens as compared to the vaccines containing OMP and LPS alone. So it was concluded that LPS no doubt act as an immunogen but its immunogenicity increases many times when combine with protein (OMP) as in whole culture vaccine. Availability: Items available for loan: UVAS Library [Call number: 2316-T] (1).

2. Genetic Diversity Among Different Isolates Of Pasteurella Multocida From Poultry

by Arslan Sardar (2013-VA-282) | Dr. Imran Altaf | Prof. Dr. Aftab Ahmad Anjum | Dr. Sehrish Firyal.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Fowl cholera is an acute bacterial disease of broiler breeders and layer breeders caused by Pasteurella multocida. In the present study, 10 isolates from different areas of Punjab were purified. These samples were confirmed by API Kit. Different molecular techniques like PCR and RFLP were used to investigate variation at the molecular level among 10 isolates collected from different areas of Punjab. Different mutations were observed among 10 field isolates at different mutation sites by sequencing. Phylogentic tree was also made using MEGA6 software that supported the sequencing results. ‘Msp1’ endonuclease cleaved bacterial whole genome at different cutting sites, all 10 isolates collected from different districts of Punjab cleaved into 3 to 5 fragments ranging from 600 to 10000 base pairs which showed the genetic variation among 10 isolates of P.mulocida. Availability: Items available for loan: UVAS Library [Call number: 2315-T] (1).

3. Assessment Of Microbial Load, Protease Activity And Aflatoxin M1 In Raw And Uht Milk Procured From Local Markets Of Lahore

by Sadaf Almas (2007-VA-250) | Dr. Imran Altaf | Dr. Ali Ahmad Sheikh | Dr. Muhammad Nasir.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Pakistan is among the largest milk producing countries. The requirement of milk is increasing day by day. It has rapidly increasing demand and competition in national and international markets. Milk consumers in Pakistan often face low-quality, lack of hygiene and absence of cold chains as primary contributors to this low quality. Milk and dairy products also become contaminated during manufacturing and packaging processes. The consumption of low quality milk may cause milk borne diseases. Not only the bacteria but the presence of their enzymes also can cause deterioration of the milk quality. The heat stable enzymes can cause the spoilage of commercial UHT products without presence of any viable count. Aflatoxin (M1) is metabolite of AFB1 in milk. It causes chronic diseases and immunosuppression in children. It is found carcinogenic and cytotoxic in nature. In this project microbial load, free amino acid estimation to predict any protease activity and Aflatoxin M1 were studied in both UHT and Raw milk samples (n=15) procured from local markets of Lahore. Three UHT brands A, B and C were purchased. The UHT milk was studied for microbial growth and protease activity at purchase and at expiry of the products. The microbial load was evaluated by testing of milk for Total viable count, Coliforms, Yeast and Molds, Anaerobic Clostridia and Bacillus cereus in both raw and UHT milk. Protease activity was estimated by assessing the free amino acid by using ninhydrin assay while the Aflatoxin M1 was detected through High performance liquid chromatography. SUMMARY 80 CONCLUSION: It was found that the locally available raw milk quality was poor for consumption and dairy processing for safe and stable milk products. UHT milk quality was found better with low microbial load. Protease activity with reference to free amino acid was detected in raw milk which is indication of the poor milk storage conditions, cold chain maintenance and unavailability of fresh milk. Protease activity was also found in UHT milk and an increase in free amino acid which could be due to heat stable proteases active during shelf life of the milk brands. Aflatoxin M1 was found in majority of raw milk sample which showed the poor animal feed storage and monitoring system. Aflatoxin M1 was also found in some samples of UHT brands with high concentration which depicted that AFM1 was heat stable and it retained in the commercial UHT products as well. Availability: Items available for loan: UVAS Library [Call number: 2354-T] (1).

4. Comparative Quality Evaluation Of Raw And Pasteurized Milk

by Hafiza Saima Ghaffar (2009-VA-230) | Dr. Imran Altaf | Prof. Dr. Aftab Ahmed Anjum | Dr. Sana Ullah Iqbal.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: This particular project was designed to evaluate the overall quality of raw and pasteurized milk available at different areas of Lahore. The parameter which was checked includes microbiological analysis, adulterants, physicochemical properties and the effect of temperature on vitamin C in milk samples. Raw samples were collected from ten different towns of Lahore, whereas pasteurized milk samples belong to ten different brands. Ten samples were collected under control conditions from animals in sterilized containers. For microbiological analysis four parameters were selected including total plate count (TPC), total coliform count (TCC), total psychrotrophic count (TPSC) and total yeast and mold count (TYMC) whereas, different adulterants like adulteration test was done such as urea, starch, hydrogen peroxide, detergent or soap, sorbitol, quaternary ammonium compound, boric acid, cane sugar, sodium chloride, formalin and hypochlorite were checked by using the milk adulteration kit in QOL. Milk contains casein and whey proteins. Whey protein being added in the milk to increase its density which is considers being an adulterant. In this project whey protein was estimated in all milk samples by titration method. Physicochemical characteristics of milk are an important parameter to judge the quality of milk. These physicochemical properties include fat%, SNF%, density kg/m3, lactose%, solid/ash, protein% and pH. Physicochemical properties were evaluated mechanically by Milkoscan. Heat treatment is an important method to reduce the microbiological contamination of milk. These treatments may include pasteurization and UHT etc. During the heat treatment some of the micronutrients may deteriorate thus compromising the quality of milk. Vitamin C is among those heat labile micronutrients. Vitamin C was checked quantitatively in market and self-collected samples by using titration method. It was concluded that total plate count TPC, TCC, TPSC and TYMC of raw milk samples were above the standard value indicating the poor quality of the milk. As far as the pasteurized milk samples were concerned ninety percent of the samples showing higher values for TCC, TPSC and TYMC. Total plate counts of all self-collected raw milk from a healthy animal were found within the standard value. Counts were in range of 3.8x 103 – 8.9x103 CFU/mL of all milk samples. TPC of all self-collected raw milk from a healthy animal were found within the standard TCC were found within permissible value (102 CFU/mL .TPSC were negative for all milk samples. TYMC were in range of 2.6x101 -7.2x101 CFU/mL. Among milk samples (n=10), three samples (30%) were positive for TYMC were while remaining samples (70%) were negative and showed no growth. Physicochemical factor show that 50 percent of raw milk have low nutritional value as compared to the standards which are buffalo and cow milk contains 7.6, 4.5% fat, 3.8, 3.8 % protein, 5.1, 4.9% lactose, 0.78, 0.72% ash and 17.0, 13.9% total solid respectively. In raw milk mean of fat (%), solid not fat (%), lactose (%), Solid/ash (0%), protein(%) and pH were 4.50±0.03, 7.915±0.06, 23.05±0.055, 3.893±0.06, 3.85±0.05, and6.9±0.0.02 respectively. In pasteurized milk mean value for fat, SNF, lactose, ash, protein and pH were 3.48 ±0.13, 7.24±0.10, 3.60±0.05,0.5 ±0.06, 2.82±0.05, 7.2±0.20 respectively. Pasteurized milk is good for consumption. Different adulterant such as urea, starch, hydrogen peroxide, Sorbitol, QAC, Boric acid, Cane sugar, NaCl, Carbonate, Formalin, hypochlorite, whey protein, Added water and soap /detergents were evaluated in all milk samples. Among these adulterant water (66%) was found in majority of milk samples, followed by whey protein (15%), starch (13%), (10%) NaCl and (8%) cane sugar were detected in raw milk samples. n Pasteurized milk samples only added water (49%) and whey protein (31%) was detected. Among the raw milk samples the maximum and minimum concentration of vitamin C was observed 0.33±0.02 and 3.33 ±0.02 mg/100ml and for pasteurized milk maximum and minimum concentration of vitamin C was observed 2.54mg/100ml and 0.32 ±0.02 mg/100ml respectively. In self- collected samples the minimum and maximum concentration of vitamin C was observed 5.25±0.02 and 8.34 ±0.04 mg/100ml respectively and after pasteurization in laboratory minimum and maximum concentration of vitamin C was observed 3.48±0.04 and 5.83 ±0.02 mg/100ml respectively. These observations had showed that pasteurization treatment decreased Vitamin C quantity. Availability: Items available for loan: UVAS Library [Call number: 2536-T] (1).

5. Affect Of Temperature, Cell Density And Multiplicity Of Infection On Biological Titer Of FMDV Type “O”

by Qazi Ithram Ul Haq (2009-VA-104) | Dr. Imran Altaf | Prof. Dr. Aftab Ahmad Anjum | Dr. Muhammad Imran.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: FMD is a transmissible viral disease of animals. It is causing very highly economical loses in Pakistan and all over the world. Through vaccination FMD is being controlled in Pakistan. Inactivated virus is used in vaccines. FMD virus grows on BHK-21 cell lines. FMDV show good adoptability on these cell lines. For good and high titer FMDV needs few physical factors to grow on BHK-21 cell lines. These factors include Temperature, Cell density and Multiplicity of infection (MOI)was considered in this research. The FMDV strain “O” was grown on BHK-21 cell line. The cells monolayer was propagated for conduction of effect of these factors on the virus. The mentioned factors were studied to get optimum level of virus titer in in vitro cell lines. The effect of 35°C, 37°C and 39°C was evaluated on the virus growth. Maximum virus propagation was noted at optimum temperature 37°C. The viral concentration at 37°C was significantly (P<0.05) higher than at 35°C and 39°C. The effect of cell density was studied on the virus concentration. Flask of three different densities 25cm2, 175cm2 and 275cm2were utilized in the current study. The virus concentration in all three different densities was not significantly different (P>0.05) from each other. Another factor Multiplicity of infection (MOI)was investigated in the study. Five different volumes 10ul, 20ul, 30ul, 40ul and 50ul of the FMDV strain “O” were used to investigate the effect of factor on the virus concentration. The results revealed highest viral harvest concretion at 50ul volume with MOI of 7.1, %age of cells infected with single virus and 6.3 × 1079. The MOI at 50ul was significantly higher (P<0.05) than the other four concentration of the virus. It was concluded from the study that the optimum temperature for the maximum FMDV concentration harvest is 37°C. The density of cell has no significant effect on the growth of virus that is flask of any density may be used to grow the FMDV. Multiplicity of infection (MOI) of 14.2 give maximum SUMMARY 34 TCID50 Optimizing the conditions for the cell culture and virus cultivation helps in maximum virus harvest achievement. From the present study it may be suggested that the physical factors may be optimized for the remaining strains of FMD and other vaccine viruses to attain maximum virus grow. Availability: Items available for loan: UVAS Library [Call number: 2681-T] (1).



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