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: ; Format:
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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.
Determination Of The Hepatitis C Virus Genotyping Prevailing In The Hepatitis Suspected Patients In District Mardan,
by Suliman Qadir Afridi | Prof. Dr. Tahir Yaqub | Dr. Fariha Akhtar | Dr. Yasin Tipu.
Material type: ; Format:
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drama
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1469,T] (1).
3.
Efficacy Of Commercial Disinfectants Against The Water Contaminating Bacteria At Commercial Broiler Farms
by Mian Muhammad Salman | Dr. Aftab Ahmad Anjum | Pfor. Dr | Prof. Dr. Tahir Yaqub.
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Publisher: 2012Dissertation note: Water is an important constituent for poultry. Poor hygienic conditions of water are health hazard for poultry. Many outbreaks are caused by consuming poor quality water.. Fifty water samples from different broiler farms in and around Lahore were collected from drinkers in sterile containers. Bacterial load was assessed using total viable count and coliform count. The counts were above the threshold level (50cfu/ml for coliform and 100cfu/ml for total viable count) showing that water used at poultry farms was of low microbiological quality. Five Disinfectants PHMB20% (.75ml/lit, 1.5ml/lit, 3.0ml/lit), PHMB11% (1.5ml/lit, 3.0ml/lit, 6ml/lit), 0.2% chlorine dioxide (0.1ml/lit , 0.2ml/lit, 0.4ml/lit) Glutral 9.8%(1.5ml/lit, 3.0ml/lit, 6ml/lit), organic acid(1.5ml/lit, 3.0ml/lit, 6ml/lit) were used and they resulted in log reduction of TVC by PHMB20% (5.83±4.36, 6.14±3.98, 9.35± 0.68), PHMB11% (9.42±0.21), 0.2% chlorine dioxide (2.45±0.97, 3.19±0.73, 6.33±0.80 ) Glutral 9.8%(6.87±1.00, 9.73±1.00,9.73±1.00), organic acid(4.75±1.21, 6.62±1.26, 6.90±1.15).PHMB20%,PHMB11%, Glutral 9.8% and organic acid were effective at normal dose, while 0.25 chlorine dioxide was effective at normal dose against at normal dose. Log reduction in Coliform count at half, normal and double dose by PHMB20% (6.52±3.33, 6.96±2.46, 7.96±0.98), PHMB11% (7.89±1.01), 0.2% chlorine dioxide (3.65±0.73, 5.08±0.98, 6.27±0.97) Glutral 9.8%(8.48±0.99), organic acid(5.18±1.21, 5.93±1.26,6.46±1.15±) . PHMB20%, PHMB11%, 9.8% Glutral, organic acid were effective against coliform bacteria at half dose while 0.2% chlorine dioxide was effective at normal dose. Glutraldehyde was effective at normal dose amongst all disinfectants against Total viable bacteria and coli form bacteria.
Availability: Items available for loan: UVAS Library [Call number: 1482,T] (1).
4.
Detection Of Soulfonamide Residues With Associated Histopathological Findings In The Tissues Of Cattle An Buffalo
by Mujahid Iqbal | r. Muhammad Yasin Tipu | Dr. Asim Aslam | Prof. Dr. Tahir Yaqub.
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print
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1483,T] (1).
5.
Sources Of Salmonella Contamination In Poultry Meat During Processing And Its Resistance To Antibioties
by Atif Masood Ahmad Khan | Prof. Dr. Tahir Yaqub | Prof. Dr. Khushi Muhammad | Veterinary and Animal.
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Publisher: 2013Dissertation note: The unhealthy birds which are slaughtered at poultry retail shops may be transferring the pathogen to the healthy meat via butcher's block, clothe (used for cleaning carcass), weighing scale, table, knife and drum in which they are bled after slaughter. The tools which are used in butchers' chicken sale point ; different objects in their shop; the feed they give to birds and bird droppings ; all were analyzed and not surprising they were found heavily contaminated with Salmonellae. Salmonellae is an enteric organism and at chicken sale points contamination to objects through birds' intestine is not much surprising. Therefore a strong need to push the pressure on government to devise laws and set standards for clean premises at chicken sale points. Extensive and irrational use of antimicrobials in human and veterinary sector in the treatment, prophylaxis and as feed additive have made this organism resistant to many of the commonly used antimicrobials.These resistant organisms are being transferred to human body due to the consumption of contaminated poultry meat. Therefore the Salmonellae in humans show resistance to many antibiotics. It is assumed that Salmonellae can transfer resistant genes via bacterial conjugation, transformation and transduction.As a result it is becoming resistant to many antimicrobials. Therefore a strong check on irrational use of antibiotics is needed.
The purpose of current study was to estimate the prevalence of antimicrobial resistant Salmonellae at chicken sale points in Lahore city. In current study 250 samples of 8 different types were collected from different poultry meat sale points in Lahore city.The selection of sale points was random. The samples included50 samples of each poultry feed and bird droppings. 150swab samples of butcher wooden blocks, table, drum( in which the birds are slaughterd), butcher's balance, knife , cloth (used to clean block, knife, table and chicken meat) were also collected from different chicken meat sale pointsin Lahore city.The Samples were analyzed for the presence of Salmonellae by culturing and biochemical tests.The percentage of Salmonellae positive samples inwooden block, weighing balance, cloth, birds' droppings, drum, bird feed, knife and table surface was 44, 24, 36 ,16, 32, 8, 28, 20 respectively.Overall prevalence of Salmonellae was 23.2 %. The isolated Salmonellae were then checked for antimicrobial resistance against 18 antimicrobials by using disk diffusion method. All the Salmonellae isolates were resistant to atleast four antimicrobials. 49 different antimicrobial resistance patterns were found.
Availability: Items available for loan: UVAS Library [Call number: 1563,T] (1).
6.
Characterization Of Mycoplasma Gallisepticum Isolates And Their Use In The Production Of Indigenous
by Mushtaq Ahmad | Prof. Dr. Masood Rabbani | Prof. Dr | Prof. Dr. Tahir Yaqub.
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drama
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1564,T] (1).
7.
Designing The Small Interference Rna Against Expression Of Coat Protein (Cp) Gene Of Potato Virus X (Pvx)
by Shafique Ahmed | Prof. Dr. Tahir Yaqub | Dr. Muhammad Wasim | Ms. Faiza.
Material type: ; Format:
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drama
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1605,T] (1).
8.
Molecular Epidemiology Of Subclinical Tuberculosis In Peri-Urban Human Population Of Lahore.
by Sadeem Shahzad | Dr. Muhammad Yasir Zahoor | Dr. Muhammad | Prof. Dr. Tahir Yaqub.
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Publisher: 2013Dissertation note: Tuberculosis (TB) is known to be a major health problem worldwide causing disease among millions of people every year. Major cause of tuberculosis in human is the infection with M.tuberculosiswhich usually causes pulmonary or lungs TB but an unknown number of patients are also infected with M.bovis which causes tuberculosis in humans as zoonotic agent along with its major hosts like cattle and deer. In developing countries where raw milk is used without pasteurisation there is a heavy risk of tuberculosis infection with M.bovis. TB infection with M.bovis mainly appears as extra pulmonary tuberculosis with and without specific symptoms of the disease.Diagnosis of subclinical asymptomatic tuberculosis and that of extra pulmonary tuberculosis is a difficult task and most of the time disease remains undiagnosed or misdiagnosed due to the unavailability of specific and sensitive diagnostic tool to diagnose the disease at early stage. Moreover prevalence of M.bovisinfection is not properly known.
This study was designed to measure the diagnostic value of Interferon gamma release assay (IGRA) for early and reliable diagnosis of subclinical extra pulmonary TB along with the molecular epidemiology of subclinical extra pulmonary TB to check the prevalence of M.bovisinfection. IGRA is a latest blood test with high specificity and sensitivity based on the principle of Interferon gamma released by effector T-Cell when exposed to M.tuberculosis antigens like ESAT-6 and CFP-10 in controlled in-vitro conditions. Eighty patients were selected for the study on the bases of the history of having day to day cattle contact along with feelings of sickness. Biopsy tissue samples of all the patients which were positive with IGRA were requested, however 24 out of 27 positive samples were collected and were first examined histologically. Twenty seven samples out of eighty were found positive with IGRA while 22 out of 24 samples were confirmed by histological examination as infected with MTB.
Both IGRA and histological examination are unable todifferentiate between the specie specific infection with M.tuberculosis orM.bovis for which differential amplification of specific fragments of bothof the species was done by running a multiplex PCR using M.tuberculosis specific 185 bp pncA product and M.bovis specific 500 bp segment. Genomic DNA was extracted from previously formalin fixed paraffin embedded (FFPE) tissues which requires pretreatment for deparaffinization. Xylene was used as deparaffinization agent. All of the twenty two samples positive with IGRA and histological study were found positive for M.tuberculosis infection and none of the sample was found positive for M.bovis infection. Results showeda close correlation among all three techniques with their specific benefits and limitations.
Study concluded that T.Spot TB (IGRA) is a potentially reliable test for the diagnosis of subclinical, extrapulmonary TB.Formalin Fixed Paraffin Embedded (FFPE) tissues may be used for TB diagnosis and other DNA based researches. Duplex PCR is a reliable technique for differential diagnosis of infection with different species of MTB complex, though none of the sample was found positive for M.boviswhich is may be due to small sample size of the study and it may further be studied in future researches. The research findings will help the clinicians to depend on IGRA testing for timely and reliable diagnosis of extrapulmonary subclinical tuberculosis and potential use of FFPE tissue samples as appropriate specimen for molecular based diagnosis of TB. Further studies are however, required to check the prevalence of M.bovis infection byincreasing sample size.
Availability: Items available for loan: UVAS Library [Call number: 1621,T] (1).
9.
Gender Differentiation From Fingerprint Ridge Count In Pakistani Population
by Ahmed Fayyaz | Prof. Dr. Tahir Yaqub | Dr. Muhammad | Ms. Sehrish Firyal.
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Publisher: 2013Dissertation note: In forensic science, fingerprinting has been used for decades as an efficient tool for identification of persons linked to an illegal activity or a crime scene. Different methods for the development and analysis of the latent fingerprints have been introduced including optical, physical and chemical methods. Each method has its own importance in the development and examination of the latent prints, which are invisible to naked eye before the application of fingerprint development methods. A lot of work has been published worldwide regarding fingerprinting. It was also reported that there is a significant difference in the ridge density of males and females. Ridge count might be helpful in the gender differentiation in Pakistani population. Patent prints of 100 males and 100 females were taken on A4 size paper or card paper using pelikan black inkpad and analysis was done with the help of 10x magnification lens. The ridges were counted diagonally within a square of 5mm x 5mm. This value depicts the number of ridges per 25 mm2. Results were analyzed by using Multivariate analysis of variance (MANOVA). The results of this study are used as a helpful tool for forensic expert and law enforcement. It reveals that females have finer epidermal ridge detail than males. The degree of ridge density is used as presumptive indicator of gender of unknown print left at a crime scene. First we qualitatively examine if prints appear coarse or fine and then by quickly quantifying ridge density or ridge count in a manner similar to method described in this study. The outcomes of this study will be helpful in exoneration of innocents in different crimes.
Availability: Items available for loan: UVAS Library [Call number: 1668,T] (1).
10.
Detection And Quantification Of Dna From Saaliva From Cigarette Butts In Different Genders
by Qurra-tul-Aien | Prof. Dr. Tahir Yaqub | Dr. Abu Saeed | Dr. Muhammad Imran.
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print
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1675,T] (1).
11.
Isolation And Characterization Of Auxin Producing Bacterial Strains From Plant Rhizosphere
by Kanwal Aziz | Dr. Jawad Nazir | Dr | Prof. Dr. Tahir Yaqub.
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Publisher: 2013Dissertation note: Auxins are a class of plant hormones or plant growth substances. Auxins have a cardinal role in the regulation of many growth and developmental processes in the plant. Soil samples were serially diluted and screened for auxin production by Salkowski method. Isolates that have the ability to produce auxin were identified by culture characters, morphology, and bio-chemical profile. From 150 isolates, 04 bacteria were selected (AUX-36, AUX-53, AUX-137, and AUX-142). The bacteria were identified by following the flowcharts described in “Berges Mannual of Determinative Bacteriology”, 9th addition. These isolates were identified as Bacillus megaterium, Escherichia coli, Klebsiella pneumonia and Bacillus marinus respectively. Next, different physical and chemical parameters for growth of bacteria and auxin biosynthesis were optimized. For the optimization of bacterial growth OD values of the culture broth (at wavelength 600nm) was taken by spectrophotometer. To estimate the amount of auxin (ìg/ml) produced in the culture broth; a standard curve (concentration of auxin ìg/ml at x-axis and OD value at y-axis) was prepared by using commercially available auxin.
The optimum conditions for growth and auxin production by AUX-36 was found to be pH 7, 0.98% osmotic pressure at 37 °C after 72 hours of incubation. If the medium is supplemented with 0.1 and 1.0% glucose, sucrose and peptone then it increased the bacterial growth which ultimately increased the auxin concentration in the broth medium. The growth and auxin
Production by AUX-53 was 0.98% NaCl concentration at 37 °C after 72 hours of incubation. The optimum pH was found to be 7 but it showed good growth at acidic as well as alkaline pH. The addition of glucose and sucrose in the growth medium increased the growth as well as auxin production. The optimum conditions for the growth of AUX-137 were as follows: pH=7, 0.98% osmotic pressure, temperature 37 °C. However the isolate had good growth at 28 °C and 2% NaCl concentration as well. The bacterial cell density and auxin increased with incubation time up to 72 hours. The isolate produced highest concentration of auxin under the same conditions. Similarly, the cell density and auxin increased with the increasing concentration of glucose in the growth medium. Sucrose increased the auxin only in the culture filtrate. While the bacteria AUX-142 showed highest growth as well as auxin production at 42 °C after 72 hours of incubation. The optimum pH and osmotic pressure was found to be 7 and 2% respectively. The cell density and concentration of auxin increased with the increasing concentration of peptone in the growth medium. Addition of tryptophan (1-2%) increased the auxin concentration in the culture supernatant of all isolates.
Next, the seed germination test and plant pot experiment were performed of selected isolates to observe the effect of bacterial inoculation on wheat plants. In seed germination test treatment of seeds with AUX-36, AUX-53 and AUX-142 significantly increased the root length and number of root hairs as compared to non-treated seeds. In plant pot experiment comparison of various growth parameters of inoculated plants with non-inoculated plants revealed the improvement in plant growth by bacterial inoculation.
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12.
Isolation And Characterization Of Phytase Producing Microrganism From Soil
by Ghazal Aziz | Dr. Aftab Ahmad Anjum | Prof. Dr | Prof. Dr. Tahir Yaqub.
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Publisher: 2013Dissertation note: Phytase is an enzyme of great importance because it is added as a biofertilizer to soil and added in animal feed to increase the uptake of inorganic phosphorous. Phytase production is the property of plant growth promoting rhizobacteria (PGPR) that harbor in rhizosphere part of the soil. These phytase producing bacteria can be utilized as biofertilizers as and can increase the soil fertility and crop production.
Soil samples were collected and screened for the production of phytase (an extracellular) enzyme on phytase screening media (PSM). Six bacterial isolates (PHY02, PHY03, PHY06, PHY07, PHY12, and PHY30) showed distinguished clear zones (> 6mm) on PSM. Isolates were identified as Lactobacillus casei PHY02, Enterobactor intermedius PHY03, Bacillus badius PHY06, Escherichia coli PHY07, Shigella sonnei PHY12, and Klebsiella pneumonia PHY30.
Effect of physical parameters (temperatures, pH and osmotic pressure) on growth and enzyme production by selected isolates was determined. Optimum growth and production of phytae by PHY02, PHY03, PHY06, PHY07, PHY12, and PHY30 (27, 9, 19, 40, 32, and 19 IU, respectively) was at 37°C. PHY07 showed highest enzyme production, followed by PHY30 and PHY02. Isolate PHY06 showed similar growth and enzyme activity at 37°C and 42°C but it was significantly reduced at low temperature. Effect of pH on phytase production on selected isolates indicates that all isolates produces maximum amount of phytase at pH 6.5. At pH 6.5 enzyme units released by PHY02, PHY03, PHY06, PHY07,
PHY12, and PHY30, were 26, 15, 19, 41, 19, and 32 IU, respectively. Production of enzyme decreased with the increase in osmotic pressure. PHY02, PHY03, PHY06, PHY07, PHY12, and PHY30 showed optimum enzyme production (27, 15, 17, 41, 18, and 32 IU, respectively) at 1 % NaCl in PSM (Figure 1C).
Effects of carbon source on both growth and phytase production of isolates showed that PHY03, PHY06, PHY07, PHY12 had significantly higher (P<0.05) cell densities and enzyme production in glucose, while PHY02 and PHY30 had higher enzyme activity at 0.3% lactose.
Nitrogen source in growing media also effects the growth and production of enzyme. PHY02 and PHY12 had better growth and production at 0.1% peptone, while PHY07 and PHY30 had significantly higher phytase level in media modified with peptone but at higher concentration (0.3%). Addition of tryptone in growth medium significantly enhanced the growth and enzyme production by PHY03, and PHY06.
Availability: Items available for loan: UVAS Library [Call number: 1685,T] (1).
13.
Molecular Characterization Of Antimicrobial Resistance Genes In Salmonella Isolates From Poultry
by Saba Zeb Khan | Prof. Dr. Tahir Yaqub | Dr. Muhammad | Ms. Sehrish Firyal.
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Publisher: 2013Dissertation note: Salmonella is a gram negative bacteria which can cause a number of different diseases including gastroenteritis, bacteremia, and typhoid fever, with the most common being gastroenteritis, some serotypes of it are pathogenic and cause serious food poisoning in humans and major economic losses in both chicken and turkeys. The birds can be the reservoir of Salmonella species which cause food borne infections in human. Human get such infections by ingesting contaminated products. In poultry farms, Salmonella can be introduced by means of contaminated feeds, particularly those that contain animal raw materials.
Use of antibiotics in poultry has become a popular practice. Different antibiotics like tetracycline, streptomycin, trimethoprim etc. are given in poultry via water and feed for growth and protection against diseases. Extensive and uncontrolled use of antibiotics resulted in increased development of antibiotic resistant bacteria. Statistical data shows that Salmonella is resistant to many antibiotics especially tetracycline.
The goal of our study was Molecular characterization of tetracycline resistance genes in Salmonella spp. and to check the prevalence of tetracycline resistance genes in Salmonella isolates from poultry drinking water.
Total 50 water samples were collected from different poultry farms and poultry meat shops in Lahore district.Various biochemical tests were performed to confirm the isolated strains as Salmonella. Tetracycline resistance was examined against isolates. Plasmid DNA was extracted from these bacteria. Antibiotic resistant plasmid genes were amplified by PCR. After gel electrophoresis the resulting fragments were sequenced through genetic analyzer. After sequencing the sequence thus obtained was compared with the reported sequences of tet genes in Salmonella strains in NCBI.
It was found out that Salmonella isolates from the poultry drinking water are highly resistant to tetracycline, as 83% of the isolated Salmonella from poultry drinking water showed their resistance towards tetracycline.PCR amplification of tet genes indicated the presence of tetA gene in 100% of tetracycline resistant Salmonella, whereas 64% of the samples contained tetB gene. TetB gene was present only in combination with tetA gene. None of the sample contained tetC, tetD and tetGgene.
This study helped to find out the prevalence of antibiotic resistant genes in Salmonella isolated from poultry drinking water, which were potential threats to human being and this study will also help us in future to develop strategies to restrict the emergence of antibiotic resistant genes and their spread.
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14.
To Investigate The Morphology Of Lip Prints And Their Effectiveness In Individualization And Sex Determination
by Makhdoom Saad Waseem Ghouri | Dr. Muhammad Wasim | Dr. Abu Saeed | Prof. Dr. Tahir Yaqub.
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not fiction
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1720,T] (1).
15.
Trace Analysis Of Gun Shot Residue On Different Fabrics Using Locally Manufactured Ammunition In Pakistan
by Muneeba Butt | Prof. Dr. Tahir Yaqub | Ms. Faiza | Ms. Sehrish Firyal.
Material type: ; Format:
print
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1733,T] (1).
16.
Molecular Characterization Of Antimicrobial Resistance Genes In Salmonella Isolates From Diarrheic Calves
by Hania Zulfiqar | Prof. Dr. Tahir Yaqub | Dr. Abu Saeed | Miss. Sehrish Firyal.
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Publisher: 2013Dissertation note: A number of infectious (bacteria, viruses, parasites) and non-infectious factors cause diarrhoea in calves. Salmonella bacteria are gram-negative and belong to the family Enterobacteriaceae. Salmonella infections in calves continue to be a major problem worldwide and are responsible of causing major economical losses. To avoid the consequences of disease caused by Salmonella drugs like pencillin, tetracyclines e.g, are given to cattle but it is observed that Salmonella show resistance against these drugs after certain period of time. Salmonella is the major causative agent of calf diarrhea. The antibiotic genes against tetracycline and ampicillin are present in slamonella isolates from calves which are suffering from diarrhea.
Aim of my study was 1) Salmonella isolation and investigation of the antimicrobial resistance gene from diarrheic calves and 2) Molecular analysis of antibiotic resistance gene of isolated salmonella species. For this purpose, salmonella antibiotic resistant isolates against ampicillin and tetracycline were selected. Antibiotic resistant plasmid genes were amplified by PCR. After gel electrophoresis the resulting fragments were sequenced through genetic analyzer.
After sequencing all the sequences were viewed in Chromas Lite 2.1.1 , Sample sequences were aligned with the reference sequences obtained from NCBI by using Mega 5.05 software. Alignment results show that there is no Single Nucleotide Polymorphism found in salmonella.
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17.
Genetic And Evolutionary Characterization Of Pakistani Pigeons And Parrots Through Mitochondrial D-
by Sehrish firyal | Dr. Ali raza awan | Prof, Dr. Aftab | Prof, Dr. Tahir yaqub.
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Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1873,T] (1).
18.
Cytochrome B Gene Amplification A Novel Approach For Diagnosis Of Theileriosis In Cattle Under Field
by Muhammad Faiz rasool | Prof. Dr.Kamran ashraf | Dr.Nisar ahmed | prof. Dr. Tahir yaqub.
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print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1893,T] (1).
19.
Seroprevalence Of Bluetongue In Domestic Animals
by Farid ahmed khan | Prof..Dr. Khushi Muhammad | Ms. Sehrish | Prof. Dr Tahir yaqub.
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Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1925,T] (1).
20.
Isolation And Molecular Identification Of H9N2 Avian Influena Virus From Human In Punjab Province Pakistan
by Abdul ahad | Prof .Dr, Masood rabbani | Prof. Dr. Rana | Prof. Dr. Tahir yaqub.
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Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1926,T] (1).
21.
Genetic Evaluation Of Cyp19A1 As A Candidate Gene For Silent Estrus Behavior In Nili-Ravi Buffalo
by Sana Imran | Miss. Maryam javed | Miss.Asma | Prof. Dr. Tahir yaqub.
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Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1969,T] (1).
22.
Molecular Variability Analysis Of Mitochondrial Dna Hypervariable Region L And Ll In Four Consecutive Human Generations of Punjab
by M. Faaras iqbal | Prof. Dr. Tahir yaqub | DR. Sehrish firyal | Miss Faiza.
Material type: ; Format:
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not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2008,T] (1).
23.
Microscopic Comparison Of Human Hair Amongst Three Male Generation Of Five Castes In Punjab Pakistan
by M. Farhan khan | Prof. Dr. Tahir yaqub | Dr. Muhammad tayyab | Dr. Sehrish.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2009,T] (1).
24.
Development Of Lamp An Economical Molecular Diagnostic Tool For Avian Influenza H9N2 The Field
by Farhana Ehsan | Prof. Dr. Tahir Yaqub | Dr.M. Imran | Ms. Faiza Masood.
Material type: ; Format:
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Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2124,T] (1).
25.
Production Of Laccase By Immobilized White Rot Fungi And Its Application For The Decolorization Of Textile Effluent Dyes
by Iqra Ghulam Rasool (2012-VA-579) | Ms. Faiza Masood | Dr. Muhammad Tayyab | Prof. Dr. Tahir Yaqub.
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Publisher: 2014Dissertation note: Textile wastewater effluent contains several types of dyes that are toxic, carcinogenic, and dangerous for environment (Nyanhongo et al. 2002). More than 10,000 different kinds of dyes and pigments are used in dyeing and textile industries. Approximately 8, 00, 000 tons colorant is produced annually and 10% of used dyes are enters the environment in the form of wastes. There are different types of textile dyes such as direct dyes, disperse dyes, reactive dyes, acid dyes, and basic dyes. Wastewater effluents discharge from textile industries contain more than 10-15% of these dyes (Kunamneni et al. 2007). Such wastewater effluents are being discharged into water stream without or after only partial treatments, causing water pollution and negatively affecting the aquatic life. The treatment of textile wastewater effluents are of major environment concerns (Nyanhongo et al. 2002).
White rot fungi (WRF) is a wide class of fungi and it is mostly comprised of basidiomycetes, ascomycetes and lignin-decomposing fungi (Wesenberg et al. 2003). WRF are the most abundant wood degraders, and are so named because they leave a bleached appearance of the wood fibers following their attack. WRF has the ability to degrade contaminants by virtue of the nonspecific nature of its extracellular ligninolytic enzyme system (Nyanhongo et al. 2002)
The white rot fungus is also known as lignin degraders because it degrades lignin effectively due to some enzymes present in it. The important enzymes involves in degradation of lignin are following: (i) lignin peroxidase: It oxidizes both phenolics and non pheolics compounds, (ii) manganese-dependent peroxidase, (iii) laccase: It oxidises phenolic compounds and produce phenoxy radicals and quinones; (iv) glucose oxidase and glyoxal oxidase used for H2O2 production, and (v) celloulobiose quinone oxidoreductase for quinone reduction (Kunamneni et al. 2007).
Laccase (oxidoreductase, EC 1.10.3.2) belongs to polyphenol oxidases group of enzymes. Copper atoms are present in the catalytic center of enzyme so it is also known as multicopper oxidases (Baldrain et al. 2006). The molecular mass of laccase is 50–100 kDa (Couto and Toca 2006). According to the mechanism of laccase, it carries out the reduction of oxygen to water along with the oxidation of its substrate. Laccases oxidize wide range of compounds such as polyphenols, methoxy substituted phenols, aromatic diamines, and other compounds (Baldrain et al. 2006).
The substrate specificity of laccase is very wide and broad. In ortho and para substituted mono and polyphenolics substrate, it carries out reduction by removing hydrogen atom from hydroxyl group. In aromatic amines, it removes one electron and produces free radicals. These radical are able of many other reactions such as depolymerization, repolymerization, demethylation, or quinone formation. During lignin degradation, oxidation of methoxyhydroquinones followed by auto-oxidation of the methoxysemiquinones. Furthermore, formation of superoxide anion radicals undergoes more chemical reactions. The activity of laccase may be increased by using different kind of activators, such as ABTS (2, 2-azinobis (3-ethylbenzthiazoline- sulfonic acid), 1-hydroxybenzotriazole, or compounds secreted by fungi (Abadulla et al. 2000). In the presence of ABTS, the decolorization efficiency increases up to 45% (Tong et al. 2007).
Laccases have been produced from different kind of sources such as some species of fungus like white rot fungi, different kinds of bacteria, and some insects (Heinzkill et al. 1998; Diamantidis et al. 2000; Dittmer and Kanost 2010). This enzyme is widely distributed in Ascomycetes, Deuteromycetes, and Basidiomycetes, WRF is the major source for the production of laccase enzyme because this fungi is involved in metabolism of lignin (Bourbonnais et al. 1995).
There are many applications of fugal laccases such as effluent decolorization discharged from industries, degradation of pulp released from paper and pulp industries, removal of phenolics compounds from alcohols, synthesis of organic compounds, biosensors, pharmaceutical sector (Yaver et al. 2001). This enzyme can also improve animal performance, increase nutrient digestibility when added to animal feed (Sharma et al. 2013). Fungal laccases have higher redox potential of +800mV as compared to plants or bacterial laccases that’s why there are several applications of laccase in biotechnology field especially in the decolorization of dyes. Enzymes can be produce in larger amount so that laccase based decolorization techniques are advantageous to bioremediation technologies (Devi et al. 2012).
Pleurotus is a species of WRF and few laccases have been isolated, purified and cloned from Pleurotus species. However, the physiological significance of laccase produced by the white rot fungi is not known. Literature reports that mycelia culture of Pleurotus florida produces at least two laccases (L1 and L2), one of which appears to be linked with the mycelia growth of the fungus (Das et al. 1997). The L1 isoenzyme is dominantly involved in the dye decolorization process.
Submerged fermentation (SmF) is a type of fermentation in which microorganism is grow in liquid broth and enzymes and other compounds are released in the broth. This technique used free liquid substrates such as nutrients etc. The substrates are utilized quite rapidly and constantly supplemented with nutrients. In fermentation broth, microorganisms are provided with appropriate nutrients and conditions such as high oxygen concentration for the production of microorganism in order to get desired products. In this technique, mycelium formation is takes place. Mycelium formation can lead to pellet formation which hinders the diffusion of oxygen and nutrients in the medium.
In recent times, wide variety of secondary metabolites has been produced commercially by fungal fermentation. Fungi are complex microorganism that is different morphologically and structurally at different phases of their life cycles form others. It is also differ in form between surface and submerged growth in fermentation media. Nature of liquid media also effect on the growth of fungi. Different culture conditions such as temperature, pH and mechanical forces are important for fungi growth but these parameters are different for different fungi (Kossen et al. 2000).
Enzymes act like catalyst and they speed up any chemical reaction without being used up in the reaction. The uses of enzymes are advantageous due to its several characteristics and features as compared to conventional chemical catalyst. However, there are some problems that can reduce the operational life time of any enzymes. These problems includes; non-reusability of enzyme, the instability of their structure, high cost of isolation, purification and characterization and their sensitivity to harsh condition of reaction.
These objectionable limitations of enzymes may be reduced by the use of immobilized enzymes. There are mainly four procedures present for immobilization of any cell (Kunamneni et al. 2007). These procedures are following: adsorption, gels entrapment or polymer entrapment, covalent coupling, and cross-linking to insoluble matrices (Brouers et al. 1989). For immobilization different kinds of matrices, such as agar, calcium alginate beads, polyacrylamide gel, etc have been used. In order to select suitable matrix and immobilization procedure, type of the cell, type of the substrate, medium conditions and products are major factors (Prasad et al. 2005).
During immobilization, enzyme is fixed to or within solid matrix in order to get heterogeneous immobilized enzyme system. Naturally enzymes are bounded to cellular membrane in living cells for most cases so in order to get the natural form of enzyme, immobilization of the cell is done. This immobilized system stabilized the structure and activity of the enzyme for longer period of time. When enzymes are immobilized, they are stronger and more challenging to harsh environment changes. Immobilization also allows easy recovery of enzyme and also it’s multiple re-use in processes. The Michaelis constant of immobilized enzymes increased and their activity usually lowered when compared to free enzyme. When immobilization procedure applied, different structural changes introduced to an enzyme which leads to these alterations. Immobilization helps to maintain the structure, stability and activity of enzyme for longer time without being de-activated (Kunamneni et al. 2007).
Immobilization represents an attractive option to obtain enzymatic catalyst for dyes treatment. This technique provides different advantages: (i) it prevents enzyme leakage even under harsh conditions; (ii) it facilitates enzyme use in different types of reactors like packed bed, stirred tank and continuous bed; (iii) it causes stabilization of the enzyme tertiary structure, usually as a consequence of multipoint attachment of the enzyme to the support, providing enzyme rigidity. The stabilization provided by covalent bonding is usually counter balanced by partial enzyme deactivation. This negative effect can be mitigated by carefully optimizing the immobilization conditions in order to maximize the ratio between immobilized enzyme activity and activity of the primary enzyme solution (Pezzella et al. 2014).
Immobilization of laccase was extensively investigated with broad range of different techniques and substrates. Inactivation of enzyme occurs when oxidized products are absorbed on the surface of the immobilization matrix support (Kunamneni et al. 2007).
Textile industries discharged wastewater effluents comprised of toxic dyes are dangerous for aquatic life and have harmful impacts on the environment. There are different methods including physical and chemical methods which are use previously to decolorized dyes. These physical and chemical methods are quite costly, prolonged, ineffective and insecure (Shang and Chi 1996; Mechichi et al. 2006). In comparison to these methods, biological processes are quite suitable and helpful. Biological processes are less expensive, safe and take less time and effective. Biological processes used microorganisms to decolorize dyes. Laccase as an extracellular oxidative enzyme produced by white rot fungi are eco-friendly and cheap. In order to decolorize dye, three day old fermentation media is used and dyes is added in the broth. To get 70-75% decolorization in fungal culture, more than 48 hours are required. Pleurotus Species produced laccase efficiently and this laccase could decolorize malachite green dye upto 70% within 24 hours (Yan et al. 2009).
Laccases can degrade several dye structures such as phenol, polyphenols and diamines (Abadulla. et al. 2000) to degrade harmful compounds into less toxic compounds and may be helpful to reduce environmental pollution (Gianfreda et al. 1999). The specific features and mechanism of laccase helps to make it a versatile biocatalyst. Due to its versatility, it is suitable for several applications such as biopulping, biobleaching, and industrial wastewater treatment. Due to the severe environment legislation, the textile industry is trying to introduce new innovative technologies for the treatment of wastewater effluents discharged from textile industries. Laccase has potential to degrade dyes of various chemical structures so that development of techniques based on laccase seems an attractive and suitable solution in decolorizing dyes (Madhavi and Lele 2009). The decolorization and detoxification of the wastewater effluent would help to use it again and again in dying process in textile wet processing.
The major purpose of this research is to decolorize the textile effluents dyes discharged by industries after partial treatment and cause water pollution and have negative effect on aquatic life and ecosystem. It is necessary to established most effective and efficient method to produce sufficient amount of laccase enzyme through immobilized white rot fungus and then utilized it in the process of bioremediation.
Availability: Items available for loan: UVAS Library [Call number: 2208,T] (1).
26.
DNA Based Characterization Of Protease Gene From Geobacillussp.Sbs-4s
by Anam Shabbir (2012-VA-608) | Dr. Muhammad Tayyab | Ms. Huma Mujahid | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Proteases are hydrolytic enzymes responsible for the hydrolysis of proteins(Qadar et al.2004).These enzymes contribute major role in textile and leather industry,accounting 60% of the world wide enzyme market(Nascimento et al.2004).These enzymes are also being used in food ,pharmaceutical ,detergent, brewage sweet industry and as digestive additives in human and animal feed (Wilson, 2012).
Proteases are produced by microbes,animal and plants but microbial proteases are preferred due to ease in production and cheaper cost (Ningthoujam et al.2010).Microbes produce a variety of proteases according to their requirement that are specific in their function (Neurath 1999).Microbes might be involved in the production of intra or extracellular proteases.Extracellular proteases help the organism to absorb and utilize hydrolytic products from proteinious substrates in order to get energy by catabolism or to synthesize the biomolecules through anabolism reactions(Ningthoujamet al.2010).
Proteases can be classified in different ways.On the basis of cutting preferences these can be divided in to two groups:endopeptidases and exopeptidases (Barret and Mcdonald 1985).Exopeptidases are involved in hydrolysis of the peptide bond near N or C terminal whereas endopeptidases are responsible for the hydrolysis of peptide bond, with the chain, distant from the peptide ends(Motyan et al .2013).On the basis of catalytic residues in active site the proteases can be divided into six groups including glutamate,serine, therionine cysteine,aspartate and metalloproteases(Li et al.2013).
Microorganisms occupy all possible environments including habitats that provides appropriate conditions for growth(Sharma et al.2009).Thermophiles have ability to grow at highertemperature whereas other microbes fail to survive.There has been increasing interest in thermophilic bacteria because of their thermostable enzyme(Obeidat et al.2012).Hyperthermophiles can survive in extremely hot environment. Hyperthermophiles occupy the most basal positions of the phylogenetic tree of life(Bouzas et al. 2006). About 70 species of hyperthermophilic bacteria and archea has been isolated from different terrestrial, marine and thermal areas in the world.Hyperthermophiles are very divergent in their phylogeny and physiological properties.Proteolytic enzymes from hyperthermophiles are catalytically active at high temperature and they can alsoretain their catalytic activity in the presence of detergent and other denaturing substances (Stetter et al.1993).
Geobacillusis widely distributed thermophiles isolated from geothermal areas (Chalopagorn et al.2014).On the basis of16SrRNA gene sequences, Geobacillus belongs to Bacillus genetic group 5. It is phenotypically and phylogeneticallyconsistent group of thermophilicbacilli (Rahman et al. 2007).Bacillus and Geobacillus species are the dominant workhorses in industrial biotechnology. These bacteria produce a variety of extracellular enzymes, such as amylases, xylanases, proteases, phytases, carbonic anhydrases, catalases, pectinases. Bacillus and Geobacillus species hasability to grow at acidic, alkaline, neutral pH and at elevated temperature has positioned them among the most important industrial enzyme producers(Satyanarayana et al. 2012).
Geobacillus are gram-positive, rod-shaped, aerobic,endospore-forming obligate thermophiles.The growth temperature for various Geobacillus species ranges from 37 to 75 °C and pH range of 6.0 to 8.5.The members of Geobacillusare homologus to each other and share homology 99% among them(Tayyab et al.2011). The genus Geobacillusthermophilicstrains, produce a variety of thermostable hydrolytic extracellular enzymes, such as proteases, amylases, and lipases used in various industrial applications (Wiegand et al. 2013)
GeobacillusSBS-4S was isolated from a hot spring located in Gilgit, Northern areas of Pakistan.Geobacillus SBS-4S strain is Gram positive, rod-shaped bacteria and occurs in chains. That could grow at a wide range of temperature (45 to 75˚C) and pH ranging 5.5 to 9.5.Geobacillus SBS-4S produced several extracellular enzymes including amylase, protease and lipase.The comparison of the strain SBS-4S with the already reported species of genus Geobacillus showed that SBS-4S is resistant to antibiotics such as streptomycine, spectinomycin and rifampicin(Tayyab et al.2011).
Availability: Items available for loan: UVAS Library [Call number: 2242-T] (1).
27.
Efficacy Of Surface Disinfectants Against Bacterial Pathogens On Experimentally Contaminated Pathogens
by Sana Ahmed (2009-VA-241) | Dr. Jawad Nazir | Dr. Amir Ghafoor | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Surface disinfection plays a key role in the prevention, control and reduction of many communicable infections as contaminated surfaces are the major source for transmission of microorganisms. Five types of surface cleaning products (Astonish Germ Clear, Cif Floor Cleaner, Dettol Surface Cleaner, Finis Floor Cleaner and Lysol Surface Cleaner) were evaluated for antibacterial activity against three bacterial cultures (E. coli, K. pneumonia, S. aureus) through Quantitative non-porous Surface Carrier Test derived from EN 13697.
Efficacy testing was performed through surface contamination techniques. Glass slides surfaces were artificially contaminated followed by recovery of the bacteria through vortex method both before and after the application of product for 5 minutes. Each of the experiment was repeated thrice and microbicidal effect (ME) values after the application of each product were calculated. Residual antimicrobial activity of the surface cleaning products was measured by applying the working solution of disinfectant on contaminated glass surfaces. Exposure time was given to the test surfaces, after each set exposure time the surfaces were treated to recover the microorganisms. Viable count from the eluant was calculated by serial dilution spread plate method. Each of the experiment was repeated thrice to find out the residual antimicrobial effect of the disinfectant products.
ME values of the Finis Floor Cleaner ranged from 4.03 to 5.14, which was the maximum value among all surface cleaning products used against E. coli. Highest ME value against S. aureus and K. pneumonia was shown by Astonish Germ Clear. The ME values ranged from 4.99 to 5.10 against S. aureus and 5.34 to 6.99 against K. pneumoniae. Finis Floor Cleaner was proved to be of maximum efficiency against E. coli where as Astonish Germ Clear was most effective against Staph. aureus and K. pneumonia. The mean log10 CFU values recovered from disinfectant treated
Summary
49
surfaces when they were exposed to the environment for different time periods of five minutes, six hours and 24 hours are 5.62 to 7.94, 4.17 to 6.35 and 7.16 to 10.25 respectively. The results indicated that the microbial count was reduced significantly at interaction time of five minutes, then at six hours the count was further reduced by Astonish Germ Clear, Cif Floor Cleaner and Dettol Surface Cleaner i.e. these surface cleaners were able to maintain their antimicrobial activity upto six hours. When the exposure to environment further increased to 24 hours, the microbial count started to increase, hence none of the disinfectants has shown antimicrobial activity upto 24 hours. This indicates that significant microbial count can be achieved within the interaction time of five minutes to six hours. Loss of antimicrobial activity upto 24 hours is probably because the active ingredients of cleaning agents get degraded during long interaction time.
Present study emphasizes that the surface disinfection process must be repeated at regular intervals. Regular and timely use of surface cleaning agents must be considered as a crucial measure in controlling disease transmission rates. Availability: Items available for loan: UVAS Library [Call number: 2294-T] (1).
28.
Study On The Pathogenesis Of Co-Infection Of Infectious Bronchitis (Ibv) And Escherichia Coli (E. Coli) In Experimental Chickens
by Sohail Khan (2013-VA-606) | Prof. Dr. Asim Aslam | Dr. Muti Ur Rehman Khan | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Infectious bronchitis and colibacillosis are infectious diseases affecting chicken of all ages and breeds. They are of major economic importance in commercial chicken flocks, causing huge losses. As both in humans and animals it is well documented that preceding respiratory infection of virus predispose individual to bacterial infection. Moreover, mix infection in poultry which occur when different organisms simultaneously invade birds is a major threat to poultry industry causes highly epidemic debilitating disease with high mortality which eventually leads to economic catastrophe. In recent past prevalence studies of field, E. coli had been reported with high prevalence and exaggerated disease along with other respiratory pathogens, additionally IBV had also isolated from same flocks in same season. Although a plethora of pioneering work had been done on IBV and E. coli in the previous decades but still a window in time exist in revealing there co-infection. Looking to field scenario in our country, the present study was designed to study an ideal challenge model for IBV and E. coli, by reproducing the natural infection.
80 SPF day old broiler birds were arranged into four groups, (A, B, C and D). Each group was comprised of 20 birds. Group D served as uninoculated control while, Group A and B were challenged with IBV on 23rd day of trails, and Group B and C were inoculated with E. coli infection on day 26th. Birds, (n=3) from each group were slaughtered on various days post infection, gross and histopathological lesions were observed and serum samples for HI were taken, throughout experiment. Variable clinical signs were recorded in various groups. In IBV infected group, respiratory distress i.e., tracheal rales, coughing, sneezing and gasping were noted during early stages, later up to 10 days post infection watery diarrhea with ruffled feathers were observed. In mix infected group clinical signs manifested rapidly and were persistent with high severity. Gross lesions in mixed infection were more profound,
Summary
56
including; airsacullitis, tracheitis with catarrhal exudation throughout respiratory tract; severe sepicemic lesions i.e. perihepaitis, pericarditis, pneumonia and polyserositis with swollen and pale kidneys distended by urates. 5 birds died in mix infected group revealed ascites with asphyxiation of trachea with caseous exudate. While in IBV infected group lesions were mild and confined to trachea, airsac and kidneys. Mortality was high in mix infected followed by IBV in which two birds died. While in E. coli and control group mortality were not noted. Histopathological lesions in mix infected group were aggravated markedly tracheal epithelium degeneration, deciliation and sloughing; congestion, interstitial nephritis, leukocytes infiltration, tubular degeneration and necrosis while were observed. In lungs, pneumonia of peribronchiolar area and interstitium with lymphocyte and macrophages infiltration, additionally degeneration and vacuolization of hepatocytes with focal necrotic areas were also noted. In IBV and E. coli group microscopic lesions were of mild degree. GMT of both IBV and mix infected birds were high but were not significant different (P>0.05). Among the groups, statistically significant increase in FCR of birds in mix infected group was observed followed by E. coli, with IBV infected came third in the row. On the bases of these findings we might conclude that mix infection of IBV and E. coli causes severe lesions with high morbidity and mortality. Availability: Items available for loan: UVAS Library [Call number: 2306-T] (1).
29.
Genetic Characterization Of Pakistani Wild Quails Using Mitochondrial Coi Gene
by Wajiha Shakil (2012-VA-817) | Dr. Ali Raza Awan | Dr. Muhammad Yasir Zahoor | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: A latest taxonomic tool termed as DNA barcoding is being used to genetically characterize animals. DNA barcoding is helpful in determining evolutionary relationship among species. Being a small sized genome and agile enough to show rapid mutation, mtDNA has been used as a pertinent marker of molecular biodiversity. DNA barcode works as an efficient tool in food manufacturing industry, diet investigation, forensics analysis, preventing unlawful trade and felonious poaching. The aim of this study was to develop DNA barcode for genetic characterization of Pakistani wild quail. Pakistani wild quail is important due to its demand for eggs, meat production, experimental purposes and gaming as well. Japanese quail was also included in this study because this quail is excessively produced in Pakistani farms. Japanese quail is present throughout the year and is comparatively bigger in size than wild quail. It has longer lifespan; farmers can easily breed this species in farms. It is suitable in poultry due to better meat yield.
COI gene (500bp) was used as a molecular marker for identification at species level. DNA was extracted from blood samples of ten wild quails (Coturnix coturnix and fifteen japanese quails (Coturnix japonica). Reported bird universal primers were used to amplify COI region from the extracted mtDNA samples using PCR. Amplicon were then sequenced by Sanger sequencing method (Sanger et al. 1977). Forward and reverse DNA Sequences were aligned with the reference sequence using nucleotide BLAST on NCBI to observe the dissimilarity among the sequences. Consensus sequences generated were used to construct their phylogenetic tree to see their evolutionary relationship with other bird species. Japanese quail which is thought to be domesticated from Japan, its Pakistani population showed close relation with sequences
Summary
90
generated in Japan for this particular species. Pakistani wild quail species showed its closest linkage with C. coturnix.
In conclusion, COI barcode proved as an authentic tool for species identification and phylogenetic inference of Pakistani wild and farm grown quails. Wild quail species has been characterized using partial COI gene sequences. This study has provided a specific genetic marker which can differentiate Japanese quail from wild quail at molecular level as most of the time both species are confused with each other. It can be helpful to the farmers and bird fanciers because they can select the birds of their choice correctly. This is the first study reporting DNA barcode of this Pakistani quail species. It would help researchers to study about phylogenetic and taxonomic status and also assist quail fanciers and quail farmers to unaffectedly identify their species of interest in farming. Identification of quail species is also important for conservation of biodiversity as it helps in preservation and identification of endangered species by generating their barcodes from even minimal evidence available. Availability: Items available for loan: UVAS Library [Call number: 2311-T] (1).
30.
Mutation Analysis Of Alpha-Synuclein Gene In Patients With Parkinson Disease
by Iffat Aleem (2009-VA-566) | Dr. Asif Nadeem | Prof. Dr. Tahir Yaqub | Ms. Huma Mujahid.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Parkinson disease is a complex, heterogeneous and chronic neurodegenerative disorder with a cumulative prevalence of greater than one per thousand, caused by neuronal loss, mainly affecting dopaminergic neurons of the substantia nigra. Parkinson disease is an idiopathic disorder of the extra pyramidal system characterized by tremors. Genetic factors contribute to its complex pathogenesis. A functional repeat polymorphism in the α-synuclein (SNCA) gene promoter conveys susceptibility for Parkinson disease. The α-synuclein (SNCA) has been implicated in rare autosomal dominant forms of Parkinson disease. The mutations in α-synuclein were associated with severe disease progression and a typical physical signs, indicative of neuro degeneration extending beyond the substantia nigra. Mutation in α-synuclein gene may have association with dopaminergic neuronal loss in Parkinson disease. Blood samples were collected from Parkinson disease patients. DNA was extracted by organic method. Primers were designed using Primer3 software. Amplification of gene was done by Polymerase Chain Reaction. PCR products were sequenced bi-directionally on ABI 3130XL Genetic analyzer. Sequence alignment was performed for polymorphism identification. The analysis of identified polymorphism has been done by CHROMAS software. Sequences were aligned by BLAST tool of NCBI. The results of analysis showed that no mutation found in exonic region of α-synuclein (SNCA) gene in Pakistani individuals selected for this study. Any change in exonic region of α-synuclein (SNCA) gene is a rare cause of sporadic and familial Parkinson disease in different populations. Availability: Items available for loan: UVAS Library [Call number: 2325-T] (1).
31.
Identification Of Genetic Variations In Toll Like Receptor 1(Tlr-1) Gene To Evaluate Its Potential For Enhanced Resistance To Bovine Tuberculosis
by Shehar Bano (2013-VA-09) | Dr. Maryam Javed | Prof. Dr. Tahir Yaqub | Miss Huma Mujahid.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Bovine tuberculosis is a disease caused by the species included in the Mycobacterium tuberculosis complex. Toll-like receptors (TLRs) are a family of conserved innate immune recognition receptors that trigger adaptive immune responses. TLR1 play an important role in host defense against mycobacteria, especially by mediating the response to mycobacterial triacylated lipopeptides.
The objective of this study is the identification of single nucleotide polymorphisms within the coding region of TLR1 gene to evaluate its potential for enhanced the resistance to bovine tuberculosis in Nili-Ravi buffalo breed. Fifty blood samples of Nili-Ravi breed were collected from UVAS Pattoki Campus, Research Farm B and Buffalo Research Institute (BRI) Pattoki. Inorganic method was used for DNA extraction, for amplification of the coding region of TLR1 gene PCR (Polymerase Chain Reaction) was used using specially designed primers and the PCR products were sequenced through Sanger’s Chain Termination method. For the analysis and alignment of sequencing the results obtained after sequencing were analyzed and aligned using the CLUSTAL W and BLAST software. After all these analysis Ten SNPs were identified in the coding region of TLR1 mentioned in table.
The Ten SNPs identified in the coding region of TOLL LIKE RECEPTOR 1 were in this order P1 C>T, P2 T>C, P3 T>C, P4 T>C, P5 T>C, P6 C >T, P7 T>C, P8 C >T, P9 A>G and P10 A>G. The one SNP found in the current research is in compliance with the (Sun et al. 2012) research on TOLL LIKE RECEPTOR 1 hence Nine SNPs found in the current research are novel in Nili Ravi buffalo. The SNPs in the exonic region that is P1 C>T, P2 T>C, P3 T>C, P4 T>C, P5 T>C, P6 C >T, P7 T>C, P8 C >T, P9 A>G and P10 A>G were all transitions i.e. the conversion of purines to purines.
Population genetic analysis and allelic distribution at all loci was analyzed using POPGENE 32 software indicated that at [P3=0.243009> 0.05] followed the assumptions of the Hardy-Weinberg equilibrium indicating that the alleles were randomly distributed throughout the population, no migration had occurred, no bottlenecks happened and population remained large in numbers.This Non-significant and obeying HWE, so can be potential marker for genetic selection.At [P1= 0.040418< 0.05], [P2=0.033603< 0.05], [P4=0.000649< 0.05], [P5=0.000262< 0.05], [P7=0.015112< 0.05] and [P9=0.000111< 0.05] the probability value below 0.05 indicated that population at these polymorphic sites was not obeying Hardy-Weinberg equilibrium. This indicated that at these positions alleles were not equally distributed in population. It can be concluded from my research that the SNPs identified in the current research may also hold potential for marker-assisted breeding programs, with the aim of breeding more BTB-resistant animals and herds within both the national farms and the private sector.
Availability: Items available for loan: UVAS Library [Call number: 2335-T] (1).
32.
Comparison Of Antifungal Activity Of Human Salivary Histatin Between Diabetic And Nondiabetic Individuals
by Farid-Ul-Haq (2013-VA-555) | Prof. Dr. Tahir Yaqub | Dr. Ali Raza Awan | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Histatins are antimicrobial proteins found in human saliva. These proteins have also been
observed to have the ability to aid in wound healing in various organisms. The genes HTN1 and
HTN3 have been studied to govern these proteins. Histatin proteins have a vast array of
antimicrobial properties. While a fungus, Candida albicans or C. albicans is a part of the human
normal gut flora, it is a threat to people who have a compromised immune system. An
overgrowth of the fungi belonging to the Candida family leads to candidiasis in humans, and oral
candidiasis has been reported to a large extent namely in diabetic patients. The antifungal
activity of histatin proteins laid the basis of the current research work.
In this study, the antifungal activity of saliva from a total of 64 healthy and diabetic
human samples against Candida albicans has been evaluated. The samples of both healthy and
diabetic human samples belong from different age ranges: 15-25, 25-35, 35-45 and 45-55 years
in order to change in antifungal activity with respect to age of an individual. Antifungal activity
was observed through both agar well and agar disk diffusion methods, with agar disk diffusion
methods showing positive results. According to the outcomes of this study at least 120μL of
healthy saliva sample is required to create a zone of inhibition. Saliva from diabetic individuals
showed no antifungal results.
This occurrence led to the next part of this study involving amplification of HTN3 gene.
The nucleotide sequences of both healthy and diabetic individuals were compared together and
showed that the absence of antifungal activity in diabetic individuals might have reasons other
than a genetic one, according to this study. The results observed from the present study indicate
that healthy human saliva possesses antifungal activity against Candida albicans. In accordance
Summary
39
to these results, the naturally occurring antimicrobial activity of histatin proteins present in
human saliva can have immense use in the field of medicine. Availability: Items available for loan: UVAS Library [Call number: 2341-T] (1).
33.
Antiviral Effect Of Human Saliva Against Avian Influenza Virus Strain H9n2
by Maryam Riaz (2008-VA-340) | Prof. Dr. Tahir Yaqub | Dr. Sehrish Firyal | Prof. Dr. Kamran Ashraf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Saliva is an important body fluid that contains a complex array of proteins, peptides and various substances that help in maintaining the health of the oral cavity. Saliva exhibits a broad-spectrum of antiviral activity against enveloped viruses as it disrupts the viral membrane. Influenza is a common virus that has been diagnosed in humans and avian species due to AIV. This study has demonstrated the naturally occurring antiviral activity of human saliva against the H9N2 influenza virus that serves as a serious threat to poultry and has been shown to possess high zoonotic potential which can cause a new pandemic.
In this study saliva samples from healthy individuals were taken and the natural antiviral ability of saliva was observed against AIV (Pk-UDL/01/08 H9N2) of calculated EID50 106.66. Inoculum prepared from saliva and H9N2 virus was injected in 9 days old embryonated eggs using CAS route and incubated at 37°C for 48 hours. A negative control (only saliva) and positive control (only virus inoculum) was also determined in the current study. The antiviral activity of saliva was observed through haemagglutination test. The HA test of harvested fluid showed that human saliva indeed possesses antiviral activity against H9N2 virus and can be used as a natural antiviral agent in medicine.
Furthermore, the genomic DNA was extracted from the blood samples. HTN3 gene responsible for histatin production, was amplified using gene specific oligonucleotides. The obtained HTN3 gene sequences were analyzed using Chromas software. The sequence alignment showed 99% similarity to the available sequences in NCBI database and 100% similarity to each individual sample. To conclude, this study has demonstrated that human saliva possesses antiviral activity against H9N2 virus. The nucleotide sequence analysis from each sample
CHAPTER 6
SUMMARY
Summary
47
showed no particular change which shows that antiviral activity of glycoproteins present in saliva does not vary at a genetic level. This innate antiviral activity can open a new frontier when it comes to combating viral infections that have grown resistant to conventional drugs in both human and animal subjects. Availability: Items available for loan: UVAS Library [Call number: 2336-T] (1).
34.
Study Of Wound Healing Effects Relating To Topical Application Of Human Saliva On Rabbits
by Sanila Amin (2013-VA-281) | Prof. Dr. Tahir Yaqub | Dr. Muhammad Imran | Dr. Habib ur Rehman.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Histatin proteins present in human saliva have been observed to show natural antibacterial and antifungal properties, as well as play a role in wound healing. These naturally occurring proteins can serve as effective agents when combating microbial infections of vulnerable wounds that have become drug resistant, without inducing negative side effects in the host. Focusing on these proteins can create a new outlook with regards to wound-healing medicine for both humans and animals.
Subjects of this study were 30 fully grown adult male rabbits weighing 2.0 to 3.4 kg and ranging from 8 to 16 months in age. They were acclimatized for two weeks in stainless steel cages and fed commercial diets, vegetables, crushed wheat and corn all over the whole experiment. Out of all 30 rabbits 24 rabbits were experimental on which saliva was applied, three were negative control to check natural wound healing, and three were positive control on which wound healing medicine was applied.
The 24 experimental rabbits were further divided into four groups with each group consisting of 6 rabbits to check the effect of age on wound healing. The age groups of human samples were divided as 15-25, 25-35, 35-45 and 45-55 (Verma et al. 2013). Saliva of human individuals belonging from these four age groups was applied on the wounds of experimental group. Furthermore, all age groups contained saliva from both gender i.e. each age group consisted of 3 male and 3 female saliva samples.
Furthermore, DNA was extracted from blood samples of the same individuals from whom saliva samples were procured. HTN1 gene which is responsible for the production of salivary histatin protein was amplified using specific primers and PCR optimization.
CHAPTER 6
SUMMARY
33
The results of this study demonstrated the wound healing properties of histatin proteins present in saliva and thus, providing a basis of using the natural ability of human saliva to act as a major component in the future of medicine for wound healing and preventing wound infections in both human and animals. Availability: Items available for loan: UVAS Library [Call number: 2344-T] (1).
35.
Detection Of Amantadine Resistant Variants Among Avian Influenza Viruses Subtype H9n2 Isolated In Pakistan
by Sabir Subhan (2009-VA-32) | Prof. Dr. Tahir Yaqub | Dr. Muhammad Zubair Shabbir | Dr. Yasin Tipu.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Avian influenza A virus subtype H9N2 is prevalent in poultry industry of Pakistan. Amantadine is an antiviral drug which is being used as prophylactic measure to control this disease despite the occurrence of resistance against this drug. There is need to monitor the resistance of amantadine in Flu viruses. In this study, we collected 100 samples of broilers birds showing mild to severe respiratory signs. Samples were collected from different locations of Punjab, Pakistan. After initial identification via Hemagglutination test, the molecular identification and confirmation of subtype H9N2 was done by multiplex PCR. To rule out the co-infection of NDV, PCR of NDV was also done. The samples which were pure H9N2 were further processed for the screening of amantadine susceptibility. To do this, titration of viruses was done on MDCK cells in the presence and absence of amantadine at the concentration of 2 ug /ml. The results of TCID5O were compared in the presence and absence of amantadine for each isolate and isolates showing difference of 2 log 10 TCID50/0.1 ml were declared resistant to amantadine as described by Masuda et al. 2000. The results of this study revealed that the viruses circulating in the poultry industry if Pakistan are resistant to this drug as we found that out 10 isolates 4 were resistant to this drug. So, there is need to monitor the usage of this drug in poultry as human cases of H9N2 viruses have been reported and virus was of avian origin. Monitoring is necessary because amantadine is recommended in flu pandemics and this virus possesses the pandemic potential and can cross the species barrier. Availability: Items available for loan: UVAS Library [Call number: 2457-T] (1).
36.
Characterization And Phylogenetic Analysis Of Neuraminidase Gene Of Avian Influenza Virus Subtype H9N2
by Muhammad Abid (2014-VA-502) | Prof. Dr. Tahir Yaqub | Dr. M. Zubair Shabbir | Prof. Dr. Aneela Zameer Durrani.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The H9N2 AIV are endemic in Pakistan since 1998 and causing serious outbreaks in poultry industry leading to increased morbidity and mortality, reduced egg production and reduced weight gain thus causing great economic losses. As these viruses have segmented genome so there is a lots of antigenic shift, antigenic drift and genetic reassortment which results in the production of new AIV subtypes. Besides causing significant losses the poultry industry, the H9N2 AIV pose a significant threat to public health and this issue has been pronounced with the fact that these viruses caused infections in Chinese children in 1999. This primary focus of this study was to characterize and to determine the phylogenetic relationship of the N2 gene of H9N2 AIV prevalent in Pakistan with other H9N2 viruses. A total of 10 H9N2 AIV were isolated from 100 samples and analyzed through serological and molecular tests. N2 gene of three isolates was amplified and sequenced.
The isolates showed 99% homology with the H9N2 AIV recently isolated from Pakistan and their phylogenetic analysis revealed that all belonged to the same G-1 lineage and fell in clusters of more recently and closely related H9N2 viruses. There were some amino acid substitutions in different positions of the NA gene as compared to previous H9N2 viruses of Pakistan and these substitutions were the same to other H9N2 viruses isolated in 2015 from Pakistan.
Due to the mutating nature of the H9N2 AIV there is need for the continuous surveillance and characterization of the prevailing H9N2 avian influenza viruses as these virus have the potential to cause serious outbreaks in poultry and also pose a significant threat to the public health. Availability: Items available for loan: UVAS Library [Call number: 2458-T] (1).
37.
Pathological Investigations Of Different Isolates Of H9n2 Prevalent In Broiler Chicken
by M. Furqan Shahid (2014-VA-322) | Dr. Yasin Tipu | Prof. Dr. Asim Aslam | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: In recent years, H9N2 virus has attained a great importance as its infection has reached panzootic proportions. AIV H9 has different antigenic variants that has made it problematic to diagnose and thus to understand the pathogenicity of this virus is also very difficult. Detection of AI H9 antibodies can be used as a complementary method for sero-epidemiological studies as an indicator of AI H9 infection. The haemagglutination inhibition (HI) assay is used routinely for subtyping and detecting an increase of antibodies to AI viruses. Surveillance and early diagnosis of AI virus is essential for poultry. It demands rapid, sensitive and inexpensive diagnostic tests. Thus, it is important to identify different antigenic variants of H9.
In this study a total of seven H9 virus samples were isolated out of total 100 collected sample from field outbreaks. These isolates were confirmed by molecular methods like PCR. Then four isolates from these seven isolates were used to infect the experimental broiler chicken. Clinical signs were recorded after the inoculation of H9N2 virus to the broiler. The results of this study showed that clinical signs were more sever upto 5 DPI. The severity of signs was proved by observing the gross pathology and histopathology of organs (Lung, Kidney, Trachea and Liver) of infected birds which were collected on 5 and 9 DPI. Serum of infected birds was also collected on 7 and 14 DPI to analyze the antibody level of infected birds against experimentally used isolate of H9N2. Then cross reactivity of different isolates of H9N2 was also checked against pannel of hyperimmune sera raised against different isolates of H9N2 and their results showed different antibody level against different isolates of H9N2. The sero-biochemical study of serum of infected birds revealed that H9N2 virus has pathogenic potential on kidney and liver.
Availability: Items available for loan: UVAS Library [Call number: 2459-T] (1).
38.
Characterization And Phylogenetic Analysis Of Hemagglutinin Gene Of Avian Influenza Virus Subtype H9n2 Isolated In 2015
by Arslan Mehboob (2009-VA-76) | Prof. Dr. Tahir Yaqub | Dr. Jawad Nazir | Dr. Maryam Javed.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: H9N2 Avian influenza outbreaks has caused great economic losses to poultry industry resulting
in decrease egg production, high morbidity and mortality. Due to different antigenic variants H9
has become problematic. It has the ability to cross species barrier and increase in pathogenicity.
Hemagglutination inhibition (HI) test is employed extensively for subtyping and detection of
antibody titre against the virus. Continuous mutations in the HA gene transforms AIV subtype
H9N2 into more pathogenic virus that may have pandemic potential and can cross species
barrier. Thus, it was necessary to identify various antigenic variants of H9 virus. It was important to
study the HA gene as it plays vital role in viral attachment, release of genetic material and
pathogenicity.
In present study, a sum of four H9 virus samples were isolated. Both serological and
molecular confirmation was done. 200 samples from different areas were collected and properly
labelled. They were then processed for egg inoculation in embryonated eggs. Virus was grown in
embryonated eggs and harvested fluid is then proceeded for confirmatory testing.
Haemagllutination and Haemagllutination Inhibition testing was done. RNA was extracted by
Kit method and cDNA was synthesized. Reverse Transcriptase (RT-PCR) was performed using
specific primer sets and then the amplicon were run on agarose gel. The bands obtained was sent
for sequencing and Phylogenetic analysis was obtained using software and tree was constructed.
Protein analysis was also performed.
The present study enabled us to characterize and construct Phylogenetic tree of HA gene of
currently prevailing H9N2 Avian Influenza isolates in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2474-T] (1).
39.
Studies on Pathogenesis and Molecular Diagnosis of Infectious Bursal Disease Virus In Broiler Chicken
by Beenish Zahid (2003-VA-134) | Prof. Dr. Asim Aslam | Dr. Yasin Tipu | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Blank CD. Availability: Items available for loan: UVAS Library [Call number: 2510-T] (1).
40.
Molecular Exploration Of Zbed6 Gene For Growth Trait In Lohi Sheep
by Usman Sagheer (2014-VA-03) | Dr. Maryam Javed | Dr. Akhtar Ali | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: ZBED6 gene is a central transcription factor. It is as a repressor of IGF2 (insulin-like growth factor II) interpretation in skeletal muscle myogenesis and development. It is essentially included in organism development, signaling, cell to cell collaboration, hepatic fibrosis, clathrin intervened endocytosis and tight intersection signaling falls. Chromatin immune precipitation (ChIP) sequencing utilizing C2C12 cells recognized around 2,500 ZBED6 binding locations in the genome, and the derived accord theme gave an immaculate match with the set up tying site in IGF2. Silencing of ZBED6 in myoblast cells influences IGF2 expression, wound healing, cell proliferation and myotube arrangement. Genes connected with ZBED6 binding sites demonstrated a very huge advancement for certain Gene Ontology groupings, including improvement and transcriptional regulation.
Forty two blood samples were collected. DNA extraction was done by using organic extraction method. Primers for PCR amplification designed using Primer3 software. PCR products were sequenced and then analyzed by using BioEdit software. Expasy translational tool for translation and POPGENE 32 software for analysis of population genetics at all the loci were used. Using this software the overall allele frequency, heterozygosity, probability using Chi-square test and Likelihood ratio test and Hardy-Weinberg equilibrium, genotype distribution at all SNP position, summary of genetic variation statistics for all loci and association were calculated. After this, for the association one way ANOVA was performed. Single nucleotide polymorphism within ZBED6 could be potential candidate gene to be serving as genetic marker for the selection of animals with higher tendencies towards weight gain. Availability: Items available for loan: UVAS Library [Call number: 2539-T] (1).
41.
Molecular Investigation Of Low Density Lipoprotein Receptor Gene Causing Familial Hypercholesterolemia And Its Evolutionary Relationship With Pan Troglodytes
by Rida Zainab (2014-VA-808) | Dr. Maryam Javed | Dr. Asif Nadeem | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Familial Hypercholesterolemia (FH) phenotype is related to improper metabolism of low density lipoproteins due to mutations in Low-density lipoprotein receptor (LDLR) gene with increased risk of ischemic heart disease. Genetic variants in LDLR gene are associated with defective catabolism of cholesterol effecting lipid metabolism which results in familial hypercholesterolemia. It occurs in both forms: Homozygous Familial Hypercholesterolemia and Heterozygous Familial Hypercholesterolemia.
Patients having high cholesterol were identified by observing the values of their serum lipid profile test reports. Their detailed history was taken and blood samples from the identified patients of familial hypercholesterolemia were collected. DNA extraction was done by Organic method. Primers were synthesized and PCR was conducted using optimized recipe and conditions. PCR products were sequenced.
Sequenced data was analyzed using Chromas or BioEdit software. BLAST was performed and sequences were aligned individually by comparing it to the reference sequence. This showed difference in any specific position of a mutated sequence against the reference sequence. CLUSTALW aligned all the sequences together in one time. Sequences were compared with reference sequence to detect the presence of any mutation or SNPs.
SNPs were identified manually and the peaks were observed in order to determine if the genotype is heterozygous or homozygous. Statistical Analysis was done and any amino acid change due to the observed SNPs was determined by using Expasy Translate Tool. It was found that both the SNPs showed amino acid changes. In the end, homology analysis was done which showed that Homo sapiens had their LDLR gene closest to that of Gorilla gorilla gorilla. Availability: Items available for loan: UVAS Library [Call number: 2551-T] (1).
42.
Prevalence Of H9n2 In Biotic And Abiotic Factors Post Avian Infleunza Outbreak In Different Districts Of Punjab
by Iqra Mahfooz (2010-VA-301) | Prof. Dr. Tahir Yaqub | Dr. Aamir Ghafoor | Dr. Maryam Javed.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Avian influenza H9N2 is not only a potential threat to the poultry industry but it is also a disease of zoonotic importance. In the past it caused very high rate of mortality in the poultry and cause huge economic loses. Present investigations were aimed to find out the uncommon resting points of avian influenza H9N2 virus in the environment. This virus is very dangerous for the poultry industry of the country so it is important to find out the hiding places of virus so that we can stop or control the future outbreaks of virus in the poultry and minimize the economic loss of the country. To rule out the above condition a total of 150 biotic and 200 abiotic samples were collected. Refrigerated samples were processed in the Influenza laboratory. Virus isolation and propagation was done through egg inoculation technique. Presence of virus was confirmed by using Polymerase chain reaction (PCR). The biotic samples (11/150) 7.0 percent reacted positive to HA, HI and also confirmed by PCR. All the abiotic samples were found negative for any evidence of presence of avian influenza virus. This study helped us in understanding the natural reservoirs of avian influenza virus. This study design revealed the hibernation of H9N2 virus in the apparently healthy flock production of broilers. Availability: Items available for loan: UVAS Library [Call number: 2563-T] (1).
43.
Pathological Investigation And Molecular Detection Of Avian Pathogenic E.Coli Serogroups In Broiler Birds
by Muhammad Azeem Riaz (2008-VA-132) | Prof. Dr. Asim Aslam | Dr. Muti Ur Rehman Khan | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The present study was designed to identify the serogroups present in field and to study their pathological effects in experimentally infected broiler chicks. The present study was attempted to scan the rfb gene clusters in APEC predominant serotypes O1, O2 and O78 strains and to develop Multiplex PCR method for serotyping of the O-antigens. The Multiplex PCR method was used for the identification of serotypes of APEC. The second part of the study was to study the pathological lesions caused by most prevalent serogroup in experimentally infected broiler chicks.
A total of 100 tissue samples (lungs and livers) were collected from colibacillosis suspected broiler birds. Streaking was done from these samples on three different media and it was found that 80% isolates were positive on MacConkey media, 60% were positive on EMB media and 40% were found pathogenic for E.coli on Congo red media.
The colonies which were of pink color on congo red media were considered as pathogenic. DNA was extracted from these colonies by boiling method by picking single colony from each petri plate. Extracted DNA was further used for PCR to confirm the three serogroups i.e O1, O2 and O78.
The PCR results showed that 8% isolated samples were found as pathogenic as O2 strain was found dominant among all. Only two genomic DNA samples were found of O1 serogroup After confirmation of serogroups inoculum of Avian pathogenic E.coli O2 strain was prepared to experimentally infect the broiler birds. Birds were infected at the age of day 7 via intratracheal route.
Following the experimental infection of birds, they were monitored for any pathological lesions which were not present significantly while some birds were off feed, reluctant to move, head down posture and were keeping themselves in isolation.
Summary
46
Postmortem of dead birds was performed and pathological lesions were noted. Livers were found to be congested, enlarged and white fibrinous layer over liver was present. Lungs were also affected with the disease and white layer was present on lungs too. Lungs were consolidated and congested.
Histopathology of lungs and livers was performed. It was noted that there was mononuclear cells infiltration and thin fibrinous layer over liver. Thickening of the liver capsule was noted due to infiltration of mononuclear cells and there was marked congestion in hepatic portal areas and the central vein. There was atrophy of adjoining hepatic cords due to greatly distended and congested sinusoids. Besides these changes, hepatocytes in various stages of degeneration along with hemorrhages, areas of congestion and fatty changes in a few places could be seen.
There was infiltration of heterophils, severe congestion, lymphocytes and macrophages in the wall of the bronchus as well as in the peribronchial alveoli. There was marked presence of granuloma in lungs. Some birds displayed thickening of the pleura and consolidated areas covered with yellowish fibrin in lungs.
The experimental infection of avian pathogenic E.coli confirmed the hypothesis that it causes pronounced histopathological lesions in broiler birds. Availability: Items available for loan: UVAS Library [Call number: 2591-T] (1).
44.
Isolation And Molecular Characterization Of Rotavirus From Calf Diarrhea And Preparation Of Vaccine
by Nadia Mukhtar (2008-VA-718) | Prof. Dr. Tahir Yaqub | Dr. Jawad Nazir | Prof. Dr. Asim Aslam.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The main contribution of the thesis “Title” is threefold. First, rotavirus was isolated and identified from calf diarrhea samples from 10 districts in Punjab. Second, optimization of molecular diagnostics and genome sequencing was done of the positive bovine rotavirus isolates from Pakistan. And thirdly, the preparation as well as evaluation of killed vaccine against bovine rotavirus isolates was performed.
The above three objectives of this study were created due to the distribution of rotavirus all over the world as an enteric pathogen in both human as well as animal species. In developing countries where cases of malnutrition are very common in young children and animals, this virus has a special importance as an etiologic agent. It causes severe diarrhea, when accompanied with severe dehydration, leads to high rate of mortality. Among the rest of the infectious diseases present in calves, neonatal diarrhea is a dire threat as it has a major impact on economic viability. Calf diarrhea is the most important problem in dairy calves that causes more financial losses to the calf producers than any other. Although numerous etiological agents may be implicated, Rotaviral diarrhea is one of the main infections causing calves to scour between five to fourteen days of age.
The cattle and buffalo calves’ population in Pakistan is devastatingly affected by the neonatal calf diarrhea due to rotavirus outbreaks. Neonatal calf mortality varies from 8.7 to 64 per cent throughout the world accounting for 84 per cent of the total mortality in the first month of age and is particularly high in the third week. While vaccination is available for the disease, it is being imported in Pakistan from other countries. The importation of the said vaccine thus, leads
SUMMARY
117
to extra expenses for the farm managers. As mentioned above one of the aims of this study is to develop an effective vaccine against bovine rotavirus and cut down expenses for farm managers.
To fulfill the objectives proposed in this thesis, rectal swabs and fecal samples were collected from public/private sector buffalo and cattle farms from 10 districts of the Punjab: Lahore, Faisalabad, Okara, Sahiwal, Sargodha, Chakwal, Bhakkar, Bahawalnagar, Multan and Bahawalpur. The samples were selected on the basis of agro-ecological zones of the province. As sampling based on agro-ecological zones allow for better data collection for recording incidence rate of the disease. Samples (n=10) from each diarrheic and apparently healthy cattle and buffalo calves from all of the districts were collected. In this way a total of 200 samples from buffalo calves and 200 samples from cattle calves were collected for this study.
Antigen of bovine rotavirus was screened from calf feces through Direct Sandwich ELISA. Bovine rotavirus samples were further confirmed through the amplification of the VP4 and VP6 genes through Rt-PCR. Homology and phylogenetic analysis of the sequenced samples was also performed. The data gathered through this analysis was helpful in collecting important data regarding the similarities as well as differences of the bovine rotavirus strain present in Pakistani isolates when compared to local regions as well as international ones. The data is also valuable when it comes to production of effective vaccines again rotavirus. RNA viruses are known to mutate unpredictably and it is safe to assume that a particular vaccine might not work effectively against all strains of a particular virus. That’s why analysis of data pertaining to all possible BRV strains is important for creation of an effective vaccine of import quality in order to help the economy of Pakistan.
Rotavirus isolate, after adaptation on MDBK cell line, was further propagated to determine TCID50 for vaccine preparation purposes. Final dose of the vaccine was adjusted to
SUMMARY
118
approximately 3ml, containing 40% culture and 60% adjuvant. Final vaccine contained 1ml of inactivated bovine rotavirus harvested culture, 1.8ml of Montanide ISA 70, 0.2ml of PBS and 0.05% of Thiomersal sodium. Efficacy of the vaccine was checked in rabbits.
For vaccine efficacy testing twenty one month old rabbits were procured. Rabbits were reared in individual isolator units in the shed facility of Quality Operations Laboratory, UVAS, Lahore. The collected rabbits were divided into two groups, vaccinated and unvaccinated rabbit groups. Each group had 10 rabbits. One ml of rotavirus vaccine was administered intramuscularly in vaccinated rabbits group. In unvaccinated rabbits group 1ml of normal saline was injected intramuscularly. The second dose of vaccine was administered at 24 days post-vaccination of first dose. The rabbits from both groups were bled at 0, 14, 28 and 42 days post-vaccination. The antibody response of rabbits to rotavirus vaccine was determined through using Antibody detection kit. The rabbits were challenged on day 42 post-vaccination using live field strain of rotavirus having TCID50 1 × 108.5. The rabbits were observed daily up to 14 days post-vaccination for appearance of diarrheic signs. The stool samples of ELISA positive were further confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR) at least 14 days post-vaccination.
The field trials were conducted at Livestock Production Research Institute, (LPRI) Bahadurnagar, Okara. The field study was done to evaluate the prepared rotavirus vaccine for prevention of neonatal calf diarrhea. For this trial, 100 dams were selected. The dams were divided into two groups and each group consisted of 25 pregnant cows and 25 pregnant buffalos. A total of 50 dams (25 cattle and 25 buffalo) were vaccinated intramuscularly with 3ml of prepared inactivated rotavirus vaccine. The 50 remaining dams (25 cattle and 25 buffalo) were kept unvaccinated.
SUMMARY
119
The blood samples were collected for serum separation after 0, 14, 28 and 42 days post vaccination in dams. The antibody titers were measured using antibody detection ELSIA kit.
After calving, newborn calves were fed with the colostrum obtained from the vaccinated dams daily for 5 consecutive days. Similarly, the calves from unvaccinated dams were fed on colostrum from their unvaccinated dams.
The 5 calves from vaccinated and 5 from the unvaccinated dams were isolated in individual isolators. These calves were challenged orally with 1ml of live field strain of rotavirus having 1 × 108.5 TCID50 and the animals were observed for diarrheic signs for 7 days.
All of the collected data was subjected to statistical analysis of (one way) ANOVA and t-test using SPSS. The <0.05 p-value determined the significance of the results through this study.
The data collected through this study allowed for the creation of valuable inferences. According to the current results of this study, the prevalence of bovine rotavirus was shown to be 6% in Punjab. This 6% included 40% and 20% from the districts of Lahore and Faisalabad respectively. Keeping these results in mind, it is to be noted that the recorded prevalence percentage from this study is higher than the prevalence of 2% in Lahore according to a previous study done in the country. It is to be noted that while the 6% prevalence of rotavirus in Punjab detected through ELISA is lower than the prevalence of 16.83% which was detected by ELISA in diarrheic calves from pervious researches, the 12% prevalence detected by ELISA in this research is higher than the prevalence of 7.25% detected by ELISA in diarrheic calves from past data.
In the present study of this thesis it was observed that the use of killed vaccine for bovines produced more efficient immune response in calves. It also enhanced the clostral rotavirus antibody titers as compared to previous studies where the use of the same strain of modified-live virus in a commercial vaccine administered IM with or without adjuvant did not significantly
SUMMARY
120
elevate colostrum antibody titers. The results collected from the present research showed that the average antibody titers in the 25 cattle dams at 0, 14, 28 and 42 days post vaccination were 0%, 57%, 68% and 78% respectively. In a similar manner the average antibody titers in the 25 buffalo dams at 0, 14, 28 and 42 days post vaccination were 0%, 55%, 70% and 82% respectively. These results indicated
the protective maternal antibody level against the rotavirus which will be transferred passively to calves. The results indicate that vaccinated dams were able to provide passive immunity to both buffalo and cattle calves in order to provide protection against the deadly virus. Availability: Items available for loan: UVAS Library [Call number: 2570-T] (1).
45.
Molecular Epidemiology Of Mycobacterium At The Animal Human Interface And Its Co-Morbidity With Diabetes Mellitus
by Zarfishan Tahir (2011-VA-624) | Prof. Dr. Mansur-ud-Din Ahmad | Dr. Abdul Majeed Akhtar | Dr. Muhammad Hassan Mushtaq | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Tuberculosis (TB) is a common and fatal infectious disease which has afflicted mankind for several millennia. At the moment, TB is positioned at number five when it comes to the most common causes of fatality worldwide. TB is curable if it is properly diagnosed and treated. In 2015, it was estimated that 1.5 million deaths (an equivalent of 4,000 deaths per day) and 9 million new TB cases have been reported. Diabetes Mellitus is also widely distributed and estimated to affect 366 million people by 2030. The co-morbidity of DM and TB is re-emerging because of the progressive epidemiology of both diseases especially in the developing countries. Endemicity of TB and DM is growing in developing countries because of low socio-economic status and poor living conditions.
In this study, a total of 500 tuberculosis positive patients were selected under TB DOTS program from five tertiary care hospitals of Lahore. Sputum samples were collected from all the enrolled patients and smear microscopy was performed for TB confirmation. Blood samples were collected from the same patients for screening of diabetes mellitus. Sputum samples were also processed for culture and drug sensitivity on LJ medium. Molecular identification by PCR technique was carried out on all positive cultured strains and results were compared with reference strain H37RV. For DNA sequencing, PCR products were sent to Singapore where sequencing was performed by Sanger method.
Data was compiled and variables including gender, age, drug resistance and treatment history and correlation among different variables was analyzed using chi-square test and Fischer’s exact test method at P-value of ≤0.05. SPSS (Statistical Package for Social Sciences, Version 20.0) was used for statistical analysis. The count data was statistically analyzed using
SUMMARY
124
descriptive statistical tools. On screening for fasting blood sugar level, 74 (14.8%) patients were recorded as diabetics as well i.e. blood sugar level ≥ 126 mg/dl. Out of these 74 patients, 22 patients had previous history of diabetes whereas remaining 52 patients were newly diagnosed at the time of screening. The maximum distribution of TB-DM patients was found in age group > 57 years. Mean age of the group without DM was 39 years and with DM was 48 years. Coexistence of DM in TB patients was higher in males (62.2%) as compared to female study subjects. However, the gender difference is statistically non-significant (p value 0.243).
The distribution of education level revealed that out of the total participants, maximum number of patients (n=220) were illiterate and similar trend was observed in diabetic patients with 54 (73%) individuals belonging to the illiterate group of the subjects. There is statistically significant difference between existence of DM and literacy level in tuberculosis patients. Among social and behavioral risk factors in tuberculosis patients, majority of the patients were unemployed (24%) in TB-DM group. Significant correlation p value ≤ 0.05 was found between coexistence of TB-DM and tobacco use. TB cases with diabetes were known to have history of smoking with 73% (n=54) while non-smokers were 27% (n=20).
On sputum smear microscopy frequency of 3+ results showing high bacterial load, was profoundly higher i.e. 67.6% in diabetic tuberculosis patients as compared to non-diabetics which was 4.9% only. Total culture yield was 363 out of 500 sputum samples. There were 193 samples that were sensitive to all drugs, 9.4% were MDR strains (resistant to Isoniazid and Rifampicin). MDR-TB is significantly higher in TB-DM patients i.e. 13.5% as compared to 8.7% in TB only patients.
In our study, DNA sequence data for drug resistance was studied by the sequence of rpoB gene of the wild type MTB strain. Sequencing results showed mutations at various spots of rpoB gene.
SUMMARY
125
Most common mutational sites identified were at codon 531, 526 and 516 with frequency of 70%, 15% and 7.5%, respectively. Moreover, mutation sites at 512 and 574 codon had also been reported. In this study, predominantly two phylogenetic variants were identified. Majority of the isolated strains were Central Asia Strain (CAS) with a prevalence of 88.2% and rest were Beijing strain. However, attempts to find zoonosis could not be established. A total of 900 raw milk samples were also screened for M. bovis and no positive sample could be detected.
The present study emphasizes the importance of screening for DM in TB patients, which had not been done in routine. This practice may prove to be helpful in reducing the disease burden of TB patients as well as DM patients. Thus it is recommended that the screening for DM should be implemented in TB/DOTS clinics.
Emergence of Multi drug resistant Mycobacterium tuberculosis is also a serious challenge for clinicians. A very large financial implication in terms of treatment, duration of chemotherapy and spread of MDR TB strains is being faced. Treating MDR TB is more complicated than treating drug sensitive TB. Patients with MDR TB require longer, much more costly treatment and experience higher mortality rates. Such a long time to initiate the treatment is not affordable, thus there is a dire need for some rapid technique like molecular based diagnostics for MDR detection, which can provide quick results and making it possible to start treatment at earlier to minimize transmission, morbidity and mortality. Availability: Items available for loan: UVAS Library [Call number: 2710-T] (1).
46.
Comparative Genomic Study of Motor Neuron Disease in Horses and Human
by Shakeela Daud (2011-VA-534) | Dr. Muhammad Wasim | Prof. Dr. Tahir Yaqub | Dr. Abu Saeed Hashmi.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: cd not submitted Availability: Items available for loan: UVAS Library [Call number: 2810-T] (1).
47.
PREVALENCE OF MAJOR BACTERIAL AND VIRAL POULTRY DISEASES IN LAHORE DIVISION OF PAKISTAN
by Wasiq mehmood (2009-VA-420) | Prof. Dr. Tahir Yaqub | Dr. Aamir Ghafoor | Dr. Yasin Tipu.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Poultry is a huge industry as an emerging agribusiness in Pakistan that contributes a lot in GDP of country but infectious diseases contribute as a major obstacle in profitable production of poultry. There is need to study the most current scenario of major diseases together with all the parameters involved. This can help in making effective preventive measures to minimize the losses. In this study we investigated 1008 cases of poultry diseases received at GP Lab, Lahore. Sick and dead birds were received from different locations of Lahore division during November, 2015 to November, 2016. Whole year was divided in to five seasons as winter (15-Nov to 15-Feb), spring (16-Feb to 14-April), hot summer (15-April to June), hot humid summer (July to 15-Sept) and autumn (16-Sept to 14-Nov). Disease was diagnosed on the basis of flock’s history, postmortem findings, isolation and identification of pathogens using various techniques of bacteriology, virology and molecular biology along with different serological techniques. The result of this study revealed that both bacterial and viral health risks are prevailing in Lahore division of Punjab however bacterial problems are greater in number in comparison of viral infections. Prevalence of E. coli infection (Colibacillosis) was greater than any other disease which could be due to poor disinfection and cleaning of control sheds along with poor management of flocks. Avian influenza and ND shared more than 90% of viral problems throughout the year. Mean prevalence of IBD was found to be 1.46% in recent year whereas CAV and adenoviral infection remained up to negligible extent. Very few cases of CRD and necrotic enteritis were reported. Prevalence of diseases has a strong correlation with seasons with incidence highest in winter and hot summer which could be due to challenging management of flocks because of severe climate conditions. Incidence in spring was found out to be 19.46. Hot humid summer and autumn were found to be least harmful seasons with prevalence of 11.23%
Summary
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and 7.11% respectively. In order to cope up with health issues, up to date studies on prevailing poultry diseases needs to be done in upcoming years as well. Availability: Items available for loan: UVAS Library [Call number: 2822-T] (1).
48.
Study Of Tyrosine Hydroxylase (Th) Gene Sequence Variations In Association With Δ9-Tetrahydrocannabinol (Thc) Dependence
by Ali Raza (2015-VA-446) | Dr. Maryam Javed | Dr. Asif Nadeem | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Anti-Social Personality Disorder (ASPD) is ability of an individual to adopt social norms. These ASPDs are driving force in majority of criminal activities. Dopamine being important neurotranmiter of nervous system controls major behavioral traits. Low level of dopamine can be causative factor for an individual to start drug abuse to restore it because majority of drug are proven to enhance dopamine production. In this study genetic exploration of genes coding for dopamine producing enzymes. Tyrosine Hydroxylase (TH) gene was selected and its two regions (Intron 1 and exon 3) were amplified and analyzed through Sanger’s sequencing method followed by statistical analysis.
Total five SNP were recorded at locus TH1, TH2, TH3, TH4, and TH5. One insertion, three transversion and only one mutation was transition. No exonic mutation was recorded hence no change in protein structure was found. Mutations at TH1 and TH4 were found to be highly associated with addiction. Mutant “B” allele were also present but still wild “A” was most common allele in our population. TH1, TH2, TH4 have positive correlation with addiction, TH3 is correlated with Nicotine and TH5 shown protective role against nicotine. There are few genetic changes in our population that can be associated with drug addiction statistically but still there prevalence in our gene pool is very low. We can conclude on the basis of these findings that drug addiction in our population is more likely a social issue rather than genetical.
Limitations of our research was sample size. There are further possibilities for this project to investigate on mRNA level. Advanced neurobiological techniques can also be applied on subjects to analyze dopamine level in different brain regions. For genetic studies epistatic role of few other genes can also be considered for validation. Availability: Items available for loan: UVAS Library [Call number: 2858-T] (1).
49.
Genetic Variation In The Promoter Region Of Pro Inflammatory Cytokine Tnf-Α Among Hiv Infected People In Lahore
by Shahid Nawaz (2015-VA-437) | Prof. Dr. Tahir Yaqub | Dr. Arfan Ahmed | Dr. Asif Nadeem.
Material type: Publisher: 2017Dissertation note: Despite of the fact that HIV is one of the most studied virus of the present time, so far there is no legitimate cure for the treatment of HIV AIDS, and in most cases AIDS is often fatal. Immune response has always been vital to cope with any disease. In context of immune response TNF-α is mainly released by the macrophages as a pro inflammatory cytokine. Studies have shown that HIV infected persons have higher concentration of TNF-α released. A particular single nucleotide polymorphism (SNP) is observed at -308 position of the promoter region of TNF- α gene due to which TNF is categorized into TNF1 and TNF2 allele. TNF2 allele is associated with higher concentration of TNF- α which in turn is associated with HIV infection. In order to know the particular association in the population of Lahore we designed this study. The hypothesis was that SNP at -308 position of the TNF- α may be associated with HIV infection. Methodology was designed as follows: 15 HIV positive samples and 15 HIV negative samples were taken and categorized into group A and B respectively. Whole Blood samples were taken from the patients and subjected to DNA extraction using commercially available kit (Favorgen blood DNA mini extraction kit). Specific primers were used to amplify the particular region (-308) of TNF- α through PCR. A small amount of PCR products were subjected to restriction enzyme Ncol. Sequencing was done and the results were analyzed using specific bioinformatics tools such as NCBI. -308 region of the TNF-alpha was analyzed for the SNPs (TNF1 and TNF2). Collected data was analyzed through SPSS for association studies. Outcome could be as follows: The study is designed to identify SNPs at -308 position of the promoter region of TNF-α gene. It will enable us to understand the possible association between TNF- α and HIV AIDS. It will be the first ever study regarding TNF- α and AIDS in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2833-T] (1).
50.
Comparative Efficacy Of Pre And Post Exposure Prophylaxis Using Indigenous Rabies Vaccine By Im Route
by Kaneez Fatima (2010-VA-202) | Prof. Dr. Tahir Yaqub | Dr. Aamir Ghafoor Bajwa | Dr. Maryam Javed.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2017Dissertation note: Rabies is a viral zoonosis that is known to be present in more than 150 countries, including Pakistan. It is a very serious health problem especially in countries with limited resources and poor awareness. It has significant economic impact and almost 100 % mortality if not properly managed.Dogs are responsible for up to 99% of all rabies transmissions to humans. Rabies is vaccine preventable viral disease. The vaccine is very expensive and a significant factor in patient’s compliance in Pakistan especially in rural areas where the main problem exist i.e. more stray dogs and increased probability of being bitten. Availability of a cheap indigenously produced effective vaccine can be very helpful in reducing the cost and overall improvement of the rabies problem in Pakistan.
We randomly selected a total of 50 patients visiting IPH. Among them 10 of pre-exposure prophylaxis and 40 for post-exposure prophylaxis. Twenty patients of the post-exposure group were children and twenty were adults. The NIH-anti rabies vaccine was administered intramuscularly to the persons visiting the IPH for pre and post exposure prophylaxis using Essen regimen. For pre exposure patients three doses on day 0,7,28 was given and for post exposure patients five doses on day 0, 3, 7, 14, and 28 was administered. The 3ml blood was collected on day 0, 28, 60 and 90 following vaccination. Serum was examined by ELISA Kit (Bio Rad Platelia rabies II Kit) for protective antibody titer.
Pakistan is importing anti-rabies vaccine which is much costly, and sometimes unavailability is a serious concern for patients. In the present study we concluded that indigenous rabies vaccine was very effective and protective levels of rabies antibody titers was detected following vaccination in all patients of the study. By widespread utilization of this vaccine we can reduce demand of imported vaccine, thus lessen the economic burden.
Availability: Items available for loan: UVAS Library [Call number: 2875-T] (1).
