1.
Effect Of Date Palm Pollen On The Plasma And Intra-Testicular Testosterone Levels Of Male Albino Rats
by Yasir Arfat | Dr. Muhammad Tayyab | Dr. Abu Saeed Hashmi | Dr. Ali Raza.
Material type: ; Format:
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Publisher: 2012Dissertation note: Considerable evidence exists for the efficacy and safety of short courses of low
dose testosterone therapy for treating infertility and delayed puberty. This treatment is
associated with high levels of patient satisfaction. There is not yet sufficient evidence for
the routine use of other therapies. Experimentally, date extract had been shown to
increase sperm count and increase stimulating concentration of testosterone count in
guinea pigs and to enhance spermatogenesis, follicle stimulating hormone (FSH) and
luteinizing hormone (LH) in rats. Intratesticular testosterone (ITT) is thought to play a
key role in the control of spermatogenesis but is rarely measured.
The present study is therefore designed to examine the effect of date palm pollen
(DPP) (Phoenix dactylifera) on the plasma and intra-testicular testosterone levels using
male albino rat as an experimental animal with the hope that the result of this study may
pave the way for treating male infertility and delayed puberty.
Adult male albino rats were divided into two groups (control and experimental).
Experimental group were given date palm pollen (DPP) suspension in a single oral dose
of 120 mg/kg of body weight for 35 days. Where as the control were given equal amount
of distilled water. Blood samples of control and experimental groups were taken for
measurement of serum testosterone levels at day 0, 12, 24 and finally at day 36.Aanimals
were sacrificed. Testes were removed for gross and biological studies. Intra-testicular
testosterone levels were measured at the end of experimental studies.
There were no statistically significant differences in the variable of control group.
Experimental group who received DPP suspension for 35 days showed statistically significant increase in body weight, weight of paired testes, serum and intra- testicular testosterone levels as compared to control group.
Availability: Items available for loan: UVAS Library [Call number: 1411,T] (1).
2.
Bioconversion Of Wheatbran To Glucose By Gluoamylase From Aspergillus Fumigatus
by Hassan Ali | Dr. Muhammad Tayyab | Dr. Abu Saeed Hashmi | Dr. Asif Nadeem.
Material type: ; Format:
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; Literary form:
drama
Publisher: 2012Dissertation note: Background:
Glucose is produced by hydrolysis of starch. Many crops like maize, rice and wheat can be used as the source of starch. Wheat bran is an agricultural waste byproduct which can be converted to glucose using glucoamylase. Wheat bran is very cheap source for carbohydrates. It is mainly composed of carbohydrates; hemicelluloses, cellulose and starch. Glucoamylase is an enzyme that yields glucose from the nonreducing chain of amylose and amylopectin by hydrolyzing ? -1,3, ?-1, 4 and ?-1,6 linkages of starch. Glucoamylases are produced by plants, animals and microorganism. Microbes, including bacteria, yeast and fungi are major source for the production of glucoamylases. Aspergillus fumigatus is found in soil and in decaying organic matter and it has an essential role in carbon and nitrogen recycling.
Hypothesis: A. fumgiatus might be a good source for the production of glucoamylase through submerged fermentation conditions.
Parameters/Methodlogy: Aspergillus fumigatus was identified macro and microscopically. Enzyme production was measured by DNS method. The effects of different sources of carbon, phosphorous and nitrogen on glucoamylase production were also examined. In order to get the optimum production of glucoamylase, the effect of temperature, pH and incubation period was analysed separately.
Methodology: Initially the A. fumigatus was isolated and conditions were optimized for the growth and production of glucoamylase. Production of enzyme was examined by DNS method. The effects of various carbon, nitrogen and phosphorous sources were examined on the production of glucoamylase. From the present study it was concluded that maximum production of glucoamylase can be obtained from A. fumigatus using wheat bran as the substrate at pH of 4.8, temperature of 40oC with an incubation time of three days.The use of wheat bran as substrate wheat bran for the production of glucoamylase will reduce the cost for the production of glucoamylase.
Availability: Items available for loan: UVAS Library [Call number: 1509,T] (1).
3.
Bioconversion of Agriculture Waste to Lysine with UV Mutated Strain of Brevibacterium Flavum and ItsBiological Evaluation in Broiler Chicks.
by Alia Tabassum | Ms. Faiza Masood | Dr. Asif Nadeem | Dr. Muhammad Tayyab.
Material type: ; Format:
print
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1530,T] (1).
4.
Biochemical Identification Of Various Causes Of Anemia In Females From District Pakpattan
by Hafiz. Muhammad Toqeer | Dr. Muhammad Tayyab | Dr. Abu Saeed Hashmi | Mr. Muhammad.
Material type: ; Format:
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Publisher: 2013Dissertation note: Anemia is estimated to be affecting almost 600 millions people all over the globe and is regarded as deficiency in Hemoglobin concentration. The decreased amount of hemoglobin in blood could not been able to fulfill the oxygen demand of tissues in body. Keeping in view the above situation, a study was planned to investigate the various types of anemia in dist. Pakpattan.
One hundred blood samples were collected from females randomly selected from various parts of district Pakpattan. The samples were divided into two groups on the basis of age. Group A contains the patients with age between 14 to 26 years where as Group B consist of patients with age 27 to 40 years. Samples were processed in-order to estimate Complete Blood Count, serum iron level, serum ferritin levels, vitamin B12 assay and HPLC based estimation of various variants of hemoglobin.
The results demonstrated that 62% of the total female population of dist. Pakpattan was found to be anemic. Among Group A, 66.66% were anemic due to iron deficiency and 33.33% were due to chronic disease. Group B contained 59.09% anemic, out of these patients, 57.69% were anemic due to iron deficiency, 38.46% due to chronic disease and 2.27% due to deficiency of Vitamin B12. Iron deficiency was found to be the major cause of anemia that is followed by anemia due to chronic disease and Vitamin B12 deficiency. The intensity of anemia was 5% higher in young age females (Group A) as compared to the elder age females (Group B).
This work provided the information about the prevalence of various types of anemia in the population of dist. Pakpattan. The data will be helpful for developing strategy for the control of anemia in future. Further study with a large number of samples, is required throughout the country for the establishment of a data base that will be a good step to control various types of anemia.
Availability: Items available for loan: UVAS Library [Call number: 1611,T] (1).
5.
Analysis Of Medulla In Human Head Halr In Different Castes
by Summaiya Aurangzeb | Dr. Wasim Shehzad | Dr. Muhammad | Dr. Muhammad Tayyab.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1633,T] (1).
6.
Evaluation Of The Detoxification Potential Of Lactic Acid Bacteria From Curd And Whey Against Ochratoxin A In Broiler
by Afshan shabbir | Ms Huma Mujahid | Dr. Asif Nadeem | Dr. Muhammad Tayyab.
Material type: ; Format:
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; Literary form:
drama
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1657,T] (1).
7.
Biochemical & Molecular Characterization Of Locally Isolated Extremophile
by Iram Murtaza | Dr. Muhammad Tayyab | Ms. Sehrish | Ms. Shagufta Saeed.
Material type: ; Format:
print
Publisher: 2013Dissertation note: Extremophiles are microorganisms with the ability to survive under extreme of conditions. Due to their extreme stability, these microorganisms produce unique biocatalysts that have been exploited in various industrial processes. These micro-organisms are unique factories for the production of enzymes that have great potential for agriculture, textile, pharmaceutical, poultry and detergent industries.
The present study was conducted for the isolation and characterization of alkaliphile. The sampling was done from spring located in Rawat, Pakistan. Optimization of growth conditions was done by growing the microorganism at various conditions including temperature, pH and salt concentration. The microorganism was identified on the basis of biochemical characteristics as well as on the basis of 16S rRNA gene sequence. Regarding the molecular characterization, the genomic DNA was isolated from the strain and was utilized for the amplification of 16S rRNA gene. The PCR product was ligated in pTZ57R/T. The ligation mixture was utilized for the transformation of E.coli DH5-? cells. The presence of insert in recombinant pTZ57R/T was confirmed by single and double restriction with EcoR1 and Hind III which resulted in the liberation of DNA fragment. The gene sequence was utilized for the phylogenetic analysis. The microorganism was found to be Gram positive rods involved in the production of catalase, amylase, protease, enzymes and gave positive results for Mannitol, Voges Proskauer Tests while negative for citrate utilization and nitrate reduction test. 16S rRNA gene sequence analysis demonstrated that the newly isolated strain showed maximum homology with various members of genus Exiguobacterium. The newly isolated strain was declared a new member of genus Exiguobacterium and was named as Exiguobacterium UVAS-01.
Availability: Items available for loan: UVAS Library [Call number: 1706,T] (1).
8.
Molecular Identification Of Bacterial Infections In Human Spontaneous Abortions
by Zarish Noreen | Dr. Muhammad Tayyab | Mr. Akhar Ali | Ms. Faiza Masood.
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drama
Publisher: 2013Dissertation note: A miscarriage medically known as spontaneous abortion is defined as a pregnancy that ends by itself spontaneously before the fetus has reached a viable gestational age of 20 to 24 weeks. Brucellosis, Q fever and Chlamydiosis are the zoonotic diseases that are widely distributed around the world and are caused by gram negative Brucella melitensis, Brucella abortus, Coxiella burnetii, Chlamydophila pecorum and Chlamydophila abortus. The current study was carried out for the molecular detection of five zoonotic bacteria in spontaneous human abortion cases. The complete blood analysis is helpful for the early diagnosis of infections in pregnancy. In this study complete blood count (CBC) and liver function test (LFT) of all patients was carried out and it was found that hemoglobin, total leukocyte count (TLC), serum bilirubin, serum alkaline phosphate, serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT) values were found to be increased as compared to normal values which indicated the fact that these parameters may fluctuate in human abortion cases.
Similarly in the present study DNA was isolated from blood samples by adopting the procedure of Genex kit. Five sets of primers were used as described earlier for identification of bacteria (Berri et al. 2009; Bally et al. 1992). In our local population of pregnant women the risk of different bacteria was evaluated and multiplex polymerase chain reaction (m-PCR) results were analyzed to determine the presence of different bacterial pathogens in all patients. The percentage prevalence of each bacterial pathogen was calculated. The prevalence of B. abortus was found to be maximum (11.6%) while B. melitensis was not detected in any patient. However, C. burnetii and C. pecorum was found to be 3.33% each and C. abortus was found to be 6.66% respectively. In healthy females no infection was observed. Quantitative data in this study was statistically analyzed using Statistical Package for Social Sciences (SPSS version 17.0). The m-PCR assay developed in current study provides a new tool for Brucellosis, Chlamydiosis and Q fever diagnosis. The application of this assay may be helpful to control animal and human infections. The study will result in the development of a diagnosis test that can be utilized for the identification of bacterial infections at early stage of pregnancy and will be helpful to reduce the number of abortions by treatment of specific bacterial infections.
Availability: Items available for loan: UVAS Library [Call number: 1712,T] (1).
9.
Nutritional Evaluation Of Jatropha Curcas Seed Meal Toxicity With Of Without Heat And Chemical Treatments
by Nadia Nawaz | Ms. Faiza Masood | Dr. Muhammad | Dr. Muhammad Tayyab.
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drama
Publisher: 2013Dissertation note: Materials and Methods: Defatted meal was mixed with Sodium hydroxide (NaOH) and methanol. 2nd sample was mixed with Sodium hydroxide NaOH and heat 3rd defatted sample was mixed with NaHCO3 solution to form a paste cover with aluminum foil and place in autoclave at 121°C for 30 minutes .The autoclave sample was dried at 250°C for 5 hours in an oven and prepared for the determination of Antinutritional factors and tried to check the best detoxification procedure and nutritional quality of Jatropha curcas seed meal. After that prepare feed and take a trail on rats, done gross pathology and biochemical analysis of blood.
Statistical analysis: Quantitative data obtained was analyzed using one way analysis of variance technique (ANOVA) under complete randomize design mean were compared using Duncan's new multiple range tests ( DMS) the statistical significance define as P ?0.05 (Nabil et al. 2011). Costat-2003, Co-Hort, version 6.303 software was used for analysis purpose.
Output: Treatment with NaOH and heat to the Jatropha meal was the best achieve method for detoxification of that seed which enhance its nutritional value.
Availability: Items available for loan: UVAS Library [Call number: 1715,T] (1).
10.
Blood Spatter Classification As A Function Of Blood Droplet Dynamics And Their Forensic Implications
by Shahid Iqbal | Mr. Akhtar Ali | Dr. Muhammad | Dr. Muhammad Tayyab.
Material type: ; Format:
print
Publisher: 2013Dissertation note: A thoroughconsideration of blood dynamics and stain creation is avital necessityto the clarification of distinct bloodstain and the spatter patterns at the crime scene. In addition the results of experimental work including studies of falling and impacting blood droplets have been presented. The magnitude of blood stains and the amount of droppers and needles round the stain fringe relays on droplet impact velocity and droplet diameter. It is not unusual to find bloodstain patterns in a violent encounter and through proper interpretation they can provide very critical details about such an event. Accurate calculations and digital photography may estimate the release height of passive droplets, the characteristics of release surface and the forces involved in bloodshed.The spattered blood pattern is used routinely in crime scene for investigators/death scene investigators to evaluate blood spattering and blood droplet impact velocity. Examination could determine the maximum resolution of bloody spots. Four different forms of stains were produced and human blood was used with heparin as an anti-coagulant (Kargeret al.1998). The blood volumes usedwere: 10 l, 5 l, 1 l, 0.5 l, 0.25 land0.1 l. Pipette (Eppendorf) and precision syringe were used for measuring and releasing the blood spattering. Blood droplets were allowed to fall freely by hand by pressing the needle of the syringe very slowly so that drops separated from the tip of a stainless steel hypodermic needle at their own mass.Respectively,all volumeswererepeated for consecutive five times to create four forms of contact stains and spatter stains on various surfaces used in the study.The resulting stains were examined with and at the end were photographed through digital camera. The results were interpreted by applying Regression coefficient relationone way Anova and two ways Anova which showed significance statistically.It will aid the crime investigation agencies to explore the importance of blood spatter analysis in crime/ death scene investigationand to estimate the creditability of reportsdelivered by the observer, prey or a doubtful.
Availability: Items available for loan: UVAS Library [Call number: 1764,T] (1).
11.
Genetic Effect Of Leptin Gene Polymorphisms On Silent Estrus Behavior In The Nili-Ravi Buffalo
by Fatima Muccee | Ms. Maryam Javed | Dr. Muhammad Tayyab | Mr. Akhtar Ali.
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; Literary form:
drama
Publisher: 2013Dissertation note: Buffalo is a high producing animal. But to exploit its full production potential is limited due to silent heat. Silent heat leads to improper diagnosis of estrus at the time of artificial insemination that causes low fertility in buffalo. Estrus is a quantitative polygenic trait controlled by environmental factors as well as polygenes. Among all the genes controlling estrus Leptin is the potential candidate gene for estrus trait and is positioned on chromosome 4q32. It stimulates production of GnRH and with FSH it controls production of estrogen thus affecting estrus behavior. The aim of the current study was to identify the single nucleotide polymorphisms in 5 flanking sequence of exon 1 and coding region of Leptin gene and to find their association with silent estrus trait. One hundred 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 and products were precipitated and sequenced for analysis. For the analysis of sequence and to identify the polymorphism bioinformatics software FinchTV software and Bioedit software were used. The 5 flanking sequence and total 3coding regions of Leptin gene were amplified with specially designed primers. The 15polymorphic sites were observed of which one SNP was found in intron 1,9 SNPs in exon 2, 4 SNPs in intron 3 and 1 SNP in exon 3 of Leptin gene. A Bioinformatics analysis was performed with the help of "POPGENE 32" software to find the association of identified polymorphisms with silent estrus. Four SNPs were found to have significant association with silent estrus with P<0.05. SNPs were analyzed for their effect and five SNPs in exon 2 were found to be synonymous, they changed the sequence of amino acids in the Leptin protein. Population genetic analysis and allelic distribution at all loci was analysed. Out of total fifteen polymorphisms, six haplotypes were constructed on the basis of DNA sequencing of individual samples. Statistical analysis of these haplotypes was done by using SHEsis software. SignalP software was used to predict the signal peptide of the Leptin protein. Phylogenetic analysis was performed and Parsimony trees were constructed by using Mega4 Software which showed sharing of cluster by Nili-Ravi buffalo breed and cattle. This genetic characterization of Leptin gene may serve as a powerful genetic source for the development of DNA markers that can be used in association studies and for selection of animals with good heat signs.
Availability: Items available for loan: UVAS Library [Call number: 1785,T] (1).
12.
Production Of Polyhydroxybutyrate From Azotobacter Vinelandii Using Molasses And Whey As Substrates
by Samia Saeed | Ms. Asma Waris | Dr. Muhammad Tayyab | Ms. Sehrish.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Polyhydroxybutyrate (PHB) is biodegradable polyester produced in nature by microbial fermentation and it is used as thermoplastic. Azotobacter vinelandii is a bacterium that accumulates PHB as intracellular granules in response to physiological stress such as excess of carbohydrate sources and limitation of nutrients e.g. nitrogen, oxygen and phosphorus etc. PHB produced in this work have great potential be used in various industries like pharmaceutics and food industry for packaging purposes and medical field. Recent research work was conducted to produce PHB form cheap agro industrial wastes like Molasses and Whey by fermentation. Different parameters such as substrate water ratio, incubation period, volume of inoculums and pH were optimized for maximum yield of PHB.
In this study fermentation media containing whey and molasses as substrates was used to check the production of PHB from the Azotobacter vinelandii. 0.5ml of inoculum media was taken in fermentation media and then kept for incubation for 24-72 hours. After incubation, culture media was centrifuged and then sediment was used for extraction, determination and identification of PHB.
It was found that Azotobacter vinelandii in molasses contained medium gives maximum yield of PHB (mg/100mL) at 4% substrate water ratio after 48 hours of incubation period (140 mg/100mL), at 2.5 mL of volume of inoculum (204 mg/100mL), at pH 8.0 (220 mg/100mL), at 0.2% of peptone (252 mg/100mL) and 0.25% (234 mg/100mL) of yeast extract. While 4% of substrate water ratio after 60 hours of incubation (128 mg/100mL), 2.0 mL of volume of inoculum (176 mg/100mL), pH 7.0 (192 mg/100mL), 0.25% of peptone (248 mg/100mL) and 0.25% of yeast extract (240 mg/100mL) were observed to be optimum parameters for maximum production of PHB from Azotobacter vinelandii in whey based medium. Data was analyzed by means of linear regression analysis to determine R (regression coefficient), which was used to find significant differences (P?0.05) in each experiment.
Conclusion: The results of present study show that molasses and whey are economically good substrates for production of polyhydroxybutyrate (biodegradable polymer) from Azotobacter vinelandii. The results also suggest that Azotobacter vinelandii is a good potential strain for production of PHB under optimized conditions.
Availability: Items available for loan: UVAS Library [Call number: 1810,T] (1).
13.
Production And Optimization Of Thermostable Recombinant A- Amylase Ffrom Geobacillus Sbs-4S
by Sabah mansoor | Dr. Muhammad Tayyab | Dr. Tanveer | Ms.Faiza masood.
Material type: ; Format:
print
Publisher: 20130Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1845,T] (1).
14.
Isolation ,Identification And Characterization Of Phytase Producing Bacteria
by Hafsa Raiaz | Dr Muhammad Tayyab | Miss Asma Waris | Miss Saeeda | IBBT.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1865,T] (1).
15.
Production Purification And Characterization Of Alkaline Proteasefrom Aspergillus Flavus Using Agricultural
by Naheed ishrat | Miss Asma waris | Dr Waseem | Dr. Muhammad tayyab.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1872,T] (1).
16.
Optimization For The Production Of Amylase By Geobacillus Sbs-4S
by Nasreen abdul jabbar | Dr. Muhammad Tayyab | Dr. Ali Raza awan | Ms. Asma waris.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1905,T] (1).
17.
Effect Of Medicinal Plant Extracts On Genes Expression In Human Cervical Carcinoma
by Atika saeed | Dr. Muhammad Tayyab | DR | Ms. Huma mujahid.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1999,T] (1).
18.
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).
19.
Refolding And Characterization Of Thermostable Recombinant Amylase From Geobacillus Sbs-4S
by Amna jawad | Dr. Muhammad Tayyab | Dr. Sehrish | Ms. Shagufta saeed.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2027,T] (1).
20.
Evaluation Of Different Methods Of Dna Extraction From Burnt Human Tissues And Generation Of Geneticprofiles For Identification
by Anum yousaf | Dr. Muhammad Wasim | Dr. Muhammad | Dr. Muhammad Tayyab.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2081,T] (1).
21.
Effect of Ginger and Turmeric Against Cadmium Induced Hepato-Renal Toxicity in Albino Rats
by Hafiza Sajda Ashraf (2012-VA-578) | Ms.Asma Waris | Dr. Muhammad Tayyab | Dr. Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Metal compounds and metal is natural elements of all ecosystems, moving between biosphere, hydrosphere, atmosphere and lithosphere. Metal complexes are increasingly introduced in the environment and could finally accumulate in a/biotic systems (Florea et al. 2005).
Contact to heavy metals is potentially damaging particularly for those metal compounds, which do not contain any physiological function in the metabolism of cells. A heavy metal is a part of an ill-defined subset of constituents that show metallic properties, which would mostly include the some metalliods, actinides, lanthanides and transition metals. Heavy metals have a high density and atomic weight much greater at least 5 times than water. Anthropogenic basis of heavy metals, i.e. contamination, have been introduced to the ecosystem waste-derived fuels are particularly prone to have heavy metals. More than 20 heavy metals, but inorganic arsenic, lead and cadmium are of particular concern (Gornal 1949).
Although, carcinogenic and toxic effects of metals have been observed in animals and humans, and that these metals form a key part in the normal functioning of biological cells. Some necessary transition metals like manganese, iron, zinc and copper contribute in controlling a variety of signaling and metabolic pathways. On the other hand their redox properties and coordination chemistry gave them an additional advantage that these metals might escape from the control mechanism such that homeostasis, partitioning, transport and binding to the designated cell elements and they interrelate with protein sites other than those which are tailor- made for them by displacing other metals from their natural binding sites. While, this process does not take place regularly, but the toxicity of metals can lead to impairment and dysfunctioning of cells (Leonard et al. 2004).
Oxidative stress is one of the main mechanisms of heavy metal toxicity. These metals are able to interact with DNA causing oxidative worsening of biological macromolecules and nuclear protein (Chen et al. 2001).
Metals like mercury, iron, cadmium, lead, copper and nickel, have the capability to produce reactive radicals, leading to cell damage like damage to lipid bilayer, depletion of enzyme activities and DNA (Stohs 1995). Moreover, these reactive radical species comprise a broad diversity of sulfur-, oxygen, nitrogen- and carbon radicals, initiating not only from lipid peroxides, hydrogen peroxide and superoxide radical but also in chelates of proteins complex peptide and amino acid, with the toxic metals. Metals produce reactive species, which in turn can cause nephrotoxicity, hepatotoxicity and neurotoxicity in humans and animals (Chen and Sthos 1995).
Cadmium is a natural metal located in the Periodic Table of the elements between mercury and zinc and the chemical behavior of cadmium is like a Zn. There is usually a divalent cation, complexd through other constituents (e.g CdCl2). Cadmium in the soil crust around 0.1ppm (Hans 1995) frequently being found as a contaminant in Pb or Zn deposits. In Zn or Pb smelting cadmium produced as a by product. Commercially, Cd is used in batteries, galvanizing steel, lasers, ink color, television screens, cosmetics and was used as an obstacle in nuclear fission and zinc to weld seals in water pipes made of lead before 1960. In the United States, approximately 600 metric tons are produced annually and about 150 tons are imported (US 2012).
Contact of Cd in human occurs mainly through ingestion or inhalation. Absorption through the skin contact is negligible. Intestinal absorption of cadmium is greater in individuals with zinc, calcium or iron deficiency (Nordberg et al. 2007).
The main source of cadmium exposure in human is considered to be the cigarette smoking (Friberg et al. 1983). Cd levels in blood and kidney are consistently elevated in smokers than nonsmokers. Inhalation exposure due to industry can be major occupational settings for example, soldering or welding and can cause a severe chemical pneumonitis (Nordberg et al 2007).
Exposure to cadmium from getting unhygienic food (eg, shellfish, leafy vegetables, rice regions of Japan and China and organ meats,) or water (either the old tap closed Zn / CD or a long-term industrial pollution) and can produce long-term effects on health (Abernethy et al. 2010).
After absorption, Cd is transported all over the body, often linked to a sulfhydryl group of protein such as metallothionein and about 30% deposits in the kidneys and 30% in the livers, and the rest scattered throughout the body (Argonne et al 2001). Half life of cadmium in the blood was estimated 75 to 128 days. (Jarup et al 1983). As a result urine, blood and hair Cd levels are poor substitutes for body burden and primarily reflect current contact; it is also true with the other heavy metals. Urine provocation test will require the estimation of cadmium in the body (Bernhoft et al. 2012).
The toxicity of cadmium has been shown in parts of body, cadmium induces tissue damage by creation of oxidative stress (Matovic et al. 2011; Patra et al. 2011; Cuypers et al. 2010) epigenetic changes in DNA expression (Wang et al. 2012; Martinez et al. 2011; Luparello 2012) mainly in the proximal segment of the renal tubule S1 (Vesay et al. 2010) inhibition or up regulation of transport routes (Therenod et al. 2012; Wan et al. 2012; Vankerkhove 2012).
Other pathologic mechanisms comprise competition disruption of the physiologic effects of Mg or Zn (Abdulla et al. 1989; Moulis et al. 2010; Shukla et al. 1984), destruction of mitochondrial function and inhibition of heme synthesis (Schauder et al. 2010), and potentially inducing apoptosis (Cannino et al. 2009). Glutathione reduction is observed, as structural deformation of proteins due sulfhydryl groups bind to the cadmium (Valko et al. 2005). Moreover, these effects are amplified by contact with other toxic metals such as As and Pb (Whittaker et al. 2011) and may be ameliorated by Se or Zn and by factors increasing levels of Nrf2 (Wang et al. 2012; Kcwill 2012).
Medicinal plants are plants having inherent active components used to treat disease or relieve pain (Okigboet et al. 2008). In most developing countries traditional medicines and medicinal plants are used as healing agents for the maintenance of good physical condition (UNESCO 1996) and in developing countries 80% of the peoples relies on traditional medicines, usually herbal remedies, for their prime health care needs (Schmincke et al. 2003). Plants extracts and their products are used in medicines as herbal remedies and they are being used to cure diverse infections (Arekemase et al. 2011). Moreover, there has been an increased concern in the beneficial potential of medicinal plants or plant products containing antioxidant properties in plummeting free radical induced tissue injury (Gupta & Flora 2005). Plants make a vital contribution to health care. The medicinal properties of plants could be based on the antimicrobial, antipyretic, antioxidant, effects of the phytochemicals in them (Cowman 1999; Adesokan et al. 2008).
Natural antioxidants also in the form of crude extracts or their chemical ingredient are very efficient in retarding the devastating processes create by oxidative stress (Zengin et al. 2011) and the toxicity analysis of the majority of the medicinal plants are not yet fully appreciated it is usually accepted that drugs which are derivative of plant products are safer than their imitative counterparts (Oluyemi 2007).
Ginger (Zingiber officinale), is a part of the Zingiberaceae family, is a eminent spice used in your daily diet (Demin et al. 2010) and also utilized for the traditional treatment of several infirmities (Afzal et al 2001). Major components of ginger like shogaol, gingerol, diarylheptanoids and volatile oil, work as antioxidant, anti-diabetic, analgesic, antipyretic, anti-inflammatory, anti-lipid and anti-tumor (Penna et al. 2003; Kadnur et al. 2005; Islamr et al. 2008; Shim et al. 2011; Kim 2008; Wangw et al. 2009). Latest scientific research has exposed that ginger has many therapeutic such as anti-oxidant effects, a capability to restrain the formation of inflammatory complexs and direct anti-inflammatory effects (Thomson et al. 2002). Ginger extract have antioxidative features, since it can scavenge hydroxyl radicals and superoxide anion. Z. officinale was found to slow down the activity of peroxidation and lipoxygenase (Topic et al. 2002).
Another, frequently used spice of Zingiberaceae: ‘curcuma longa’ (turmeric) has shown its strong intrinsic activity as a healing agent for several ailments. The active ingerdient of turmeric is the Curcumin that (Curcuma langalinn) shows antioxidant property. It is a yellow coloured phenolic pigment yield from the turmeric rhizomes (family Zingiberaceae).The most significant characteristic of curcumin is that it has no side consequences, regardless of the therapeutic agent in a number of useful purposes. It acts as a scavenger of free radicals (Khanna et al. 1999). Curcumin is considered to be an efficient antioxidant against oxidative tissue damage. It can considerably restrain the generation of reactive oxygen species (Joe et al. 1994) Moreover, curcumin is considered to be a powerful inhibitor tumour cells proliferation (Joe et al. 2004) a powerful cancer chemopreventive agent (Duvoix et al. 2005; Aggarwal et al. 2005) an dexhibits anti carcinogenic, anti-infective and anti viral properties (Araujo et al. 2001).
Availability: Items available for loan: UVAS Library [Call number: 2199,T] (1).
22.
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.
Material type: ; Literary form:
not fiction
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).
23.
Phylogenetic Analysis Of Major Fresh Water Carps Of Pakistan Through DNA Barcoding
by Madeeha Awan (2012-VA-650) | Dr. Ali Raza Awan | Dr.Sehrish Firyal | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Pakistan is bestowed with the land of geological and topographic diversity. The ecological variation is uniformly reflected in all water lands of the country. Pakistan has significantly huge natural inland water resources in the form of ocean, rivers, networks of canals and lakes (Mirza and Rafique 1994). The country is blessed with one of the largest freshwater resources in the world correspondingly large number of freshwater living vertebrates is available from which fishes are quite significant considering the ecological balance and its consumption as food. It is one of the food sources which solely provide all the essential nutrients, minerals and high quality protein which is not common from other food items (Muhammad Rafique 2007). Out of 33,100 fish species identified worldwide as per Fish Base organization report published in April 2015 (http://www.fishbase.org). Out of 233 (indigenous and exotic) freshwater fish species, 78 economically important indigenous fish species are available in the water bodies of the Pakistan. According to this report fishes are the largest vertebrate group, constituting about 50% of all vertebrate species. Systematically fishes are widely spread in nature, ranging from prehistoric jawless fishes to cartilaginous fishes and also from old to current day bony fishes. The taxonomic placement of these fishes shows their belonging to class Actinopterygii, sub-class Teleostei, 3 cohorts, 6 superorders, 13 orders, 30 families and 86 genera (Rafique 2007; Rafique and Khan 2012).
Demand of fish is increasing day by day not only being the naturally available source of food rather the health benefits associated with its consumption. This necessitates to develop a more efficient and sustainable system to increase their growth. DNA based technologies are being competently employed in aquaculture production fields for pedigree information.
Introduction
2
Moreover, tagging each fish individually is not an easy task so these DNA based methods help in avoiding intrusion of environmental factors which may result from raising fish families in separate reservoirs (Martinez 2007). Fish identification has been traditionally based on phenotypic features. However, due to high multiplicity and morphological similarity, in many cases, fish at its different developmental stages are hard to be identified by relying only on morphological characteristics (Victor et al. 2009).
For phylogenetic studies of the animals the use of mt-DNA is very common and reliable compared to nuclear DNA due to its high evaluation capabilities, which results in gathering of differences even between closely related species (Moore 1995; Mindell et al. 1997).“Bar-coding gap" is the name given to the property that is inter-specific variation in this region is markedly higher than intra-specific variability (Hebert et al. 2003).
Approximately each and every animal contain 13 protein-coding genes (PCGs) as an essential component of their mt-genome (mitochondrial genome), which helps in encoding of several proteins responsible for the oxidative phosphorylation machinery (Richly A et al. 2004, Song H et al. 2008). Being maternally inherited, mt-DNA is better as compared to genomic DNA such as quick evolution, less exposure to recombination, high copy number, high conservation, little duplications and negligible intergenic regions (Waugh J 2007, Xu J 2005). Clonal inheritance is the main property which makes it more worthy and suitable marker in comparison with other available molecular bio-diversity tools (Galtier et al. 2009).
DNA barcoding is one of the taxonomic tools. It is being used to distinguish animal species based on the small segment of their genome such as mitochondrial DNA, designated as an identification tag or barcode of particular species (Herbert et al. 2003). Identification of species using DNA barcoding is quite debatable. Still many researchers consider it as a reliable
Introduction
3
basic tool to ascertain the genetic characterization of diverse eukaryotic species, especially after establishment of the Consortium for the Barcode of Life (CBOL) in 2004 [http: //www. barcodeoflife.org/].
Ideally DNA barcoding should provide quick, reliable and cost effective species identification, even to those user who has little or negligible knowledge of taxonomy (Herbert et al. 2003, Hajibabaei M et al. 2005, Herbert et al. 2005). Identification of unknown source is possible by using distance based tree which can be created by comparing unidentified sequences against retrieved known sequences of different species (Hebert et al. 2003, 2004a, 2004b). DNA barcoding identification system has been recognized universally as standardized method to recognize species and unveil their genetic diversity (Herbert et al. 2003; Herbert et al. 2004). The ideal DNA barcoding is robust, with conserved, universal primer binding sites, reliable DNA amplification and sequencing.
From whole mitochondrial genome, Cytb (Cytochrome b) is considered as one of the most promising gene due to its function and structure, even it is composed of both conserved and rapidly evolving regions which are more related to evolutionary studies (Farias et al. 2001). To identify unknown or ambiguous species it is considered more reliable as it contain sequences which provide the specific information about particular species (Parson W et al. 2000a, 2000b). It is also one of the most useful genetic marker to identify the linkage within families and genera (Meyer 1994; Teletchea 2009). Cytb gene is involved in comparative studies which results in development of new classification schemes and been used to assign a genus to a newly described species as well as improve the understanding of evolutionary relationships of genra (Castresana 2001).
Introduction
4
One of the core objectives of this study is to identify and classify four freshwater indigenous fish species of Pakistan, which includes Labeo rohita (Rohu), Labeo calbasu (Calbans), Catla catla (Thalla) and Cirrhinus mrigala (Mori) using Cytb gene. Morphologically, Labeo rohita shows compressed body with convex dorsal profile while mouth bears a pair of barbells and fins are gray and orange in color. Catla catla shows compressed body with broad head. Mouth is wide with thick lower lip. Labeo Calbasu`s dorsal profile is more convex than that of abdomen and two pairs of barbells are present on fringed upper lip. Cirrhinus mrigala has elongated and streamlined body shape which is grayish and silver in color (Bhuiyan AL 1964; Rahman AK 1989). All of these species are found in freshwater bodies mostly lakes, rivers and ponds except Labeo calbasu which is a bottom dweller. These fishes are harvested by using rod and line or by using nets (Talwar PK and Jhingran AG 1991). These fishes are known as major carps and economically very important for the country due to their high consumption as food. These fishes are also used for fish farming due to their greater muscle mass thus also possess very high commercial value for fish farming business.
Another objective of this study is to resolve the taxonomic anomalies related to above mentioned species. Selling of fish meat in mislabeled packaging is a serious issue now days. Most commonly Hypophthalmichthys molitrix (silver carp) and Ctenopharyngodon idella (grass carp) are sold under the label of Labeo rohita. DNA barcoding is also helpful in detecting such fraudulent mislabeling.
It would be the first study in Pakistan to genetically characterize commercially important fish species. It would help scientists to know about their phylogenetic and taxonomic status and also assist fish fanciers to genuinely identify their species of interest. Identification of fish species is also important for conservation of biodiversity as it helps in preservation and
Introduction
5
identification of endangered species by generating their barcodes from even minimal evidence available. This study has paved the way for molecular biologists to study taxonomic ambiguities at sub species level using SNP (Single nucleotide polymorphism) based identifying marker. Availability: Items available for loan: UVAS Library [Call number: 2207-T] (1).
24.
Bioconversion Of Molasses To Glucose Oxidase Through Solid State Fermentation With Aspergillus Niger
by Wajeeha Zafar (2012-VA-574) | Dr.Abu Saeed Hashmi | Dr. Muhammad Tayyab | Dr. Muhammad Wasim.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Enzymes can be defined as soluble colloidal organic catalysts which are produced by living cells and are capableof acting independently of the cells. Glucose oxidase belongs to oxidoreductaseand is also called as glucose dehydrogenase. The glucose oxidase enzyme (GOX) oxidizes glucose to gluconic acid. In cells, it aids in breaking the sugar down into its metabolites. Glucose oxidasehas found several commercial applications including glucose removal from dried egg; improvement of color, flavor, and shelf life of food materials; oxygen removal from fruit juices, canned beverages. It has also been used in an automatic glucose assay kit in conjunction with catalase and chiefly in biosensorsfor the detection and estimation of glucose in industrial solutions and in body fluids such as blood and urine. It is often extracted from Aspergillusniger. GOX is a dimeric protein. The active site where glucose binds is in a deep pocket. This enzyme acts outside of cells, is covered with carbohydrate chains (Raba and Horacio 1995).
Aspergillus niger is the potential source for the production of glucose oxidase and is preferreddue to its high production ration of extracellular enzyme. The ability of Aspergillus niger toutilize a wide range of waste products as nutrition source makes it more economical source of the enzyme (Rajesh et al .2002).
The glucose oxidase fromA. nigerisalso an intracellularenzyme present in the mycelium of the organism. Aspergillus nigeris a filamentous fungus belonging to phylum Ascomycota. It Produces microscopic conidia on conidiophores that are produced asexually. Hyphae possess septa and are hyaline. They are supported at their base by foot cells from which conidiophores originate. It possesses long, double-walled, smooth and colorless to brown conidiophores.It is commonly foundin mesophilic environments such as soil, plants and enclosed air environments. It is capable of surviving in various environments, it is not only a xerophilic fungus, but is also a thermo tolerant organism. It is because of this property that it exhibits a high tolerance to freezing temperature(Schuster 2002).
Glucose oxidase was first isolated from mycelia ofA. nigerandPenicilliumglaucumby Müller.
A large number of microbes including bacteria and filamentous fungi have been used for the production of glucose oxidase. Glucose oxidase is produced at large scale using A. nigerand P. amagasakiense. Many bacteria are also involved in the production of this enzyme; some of these are Zymomonasmobilis, Micrococcus and Enterobacte(Yogananth et al. 2012).
Glucose oxidase (GOX) from Aspergillus niger is a well-characterised glycoprotein consisting of two identical 80-kDa subunits with two FAD co-enzymes bound. Both the DNA sequence and protein structure at 1.9 A have been determined that these identical subunits size vary from 70 to 80 KDa. It catalyzes the oxidation of D-glucose (C6H12O6) to D -gluconolactone (C6H10O6) and hydrogen peroxide. It is produced naturally in some fungi and insects where its catalytic product, hydrogen peroxide, acts as an anti-bacterial and anti-fungal agent (Ikram et al . 2014).
Glucose oxidase has a molecular weight of 160,000 a.m.u. (Tsugeet al .1975) and consists of two identical polypeptide chain subunits having nearly equal molecular weights linked by disulphide bonds (O'Malley and Weaver 1972) and it is highly specific for β-D-glucose (Bentley 1963). Each subunit of the glucose oxidase contains one mole of Fe and one mole of FAD (Flavin adenine dinucleotides) and it contains 74% protein, 16% natural sugar and 2% amino sugars (Tsugeet al. 1975). The Glucose oxidase enzyme in its purest form is pale-yellow powder. The molecular weight of GOX ranges from approximately 130 kDa to 175 KDa (Kalisz et al. 1997).
Gluconic acid, the oxidation product of glucose, is a mild neithercaustic nor corrosive, non-toxic and readily biodegradable organic acid of great interest for many applications. As a multifunctional carbonic acid belonging to the bulk chemicals and due to its physiological and chemical characteristics, gluconic acid itself, its salts (e.g. alkali metal salts, in especially sodium gluconate) and the gluconolactone form have found extensively versatile uses in the chemical, pharmaceutical, food, construction and other industries (Anastassiadis and Morgunov 2007 ).
This enzyme is present in all aerobic organisms and normally functions in conjunction with catalase (Coxon and Schaffer 1971). This enzyme is also used as an antioxidant (Berg et al. 1992). It is mainly available from microbial sources and is normally produced by aerobic fermentation of Aspergillus nigerand Penicillium species (Fiedurak1996; Lu et al. 1996; Plush et al .1996; Rando et al. 1997). It has high specificity for D-glucose (Kuly and Cenas 1983).
This enzymeis also widely used to produce gluconicacidthatGOXtogether with Horse Reddish peroxidase has a range of applications in the food industry for glucose determination. GOX is being used in the textile industry producing hydrogen peroxide for bleaching process. This enzyme is also used to determine capillary glucose in screening of gestational diabetes (Mesiggi et al. 1988). This enzyme is utilized to extend the shelf life of fish(Field et al.1986) andproduction of calcium gluconate, gluconic acid and its derivatives (Khurshid2009).
Solid state fermentation [SSF] has been recently considered as the most cheapest and more environmentally friendly relative to submergedliquid fermentation [SLF] in the production of value added industrial based products such as enzymes, bio fuels.Advantages of Solid State Fermentation over Submerged Fermentation isHigher volumetric productivity, usually simpler with lower energy requirements, Might be easier to meet aeration requirements, Resembles the natural habitat of some fungi and bacteria and Easier downstream processing (Mienda et al. 2011).
Molasses is a dark brown, almost black, moist granular sugar. Its distinctive molasses taste is due to its high content of minerals. Nutritively, it has high iron content (Draycott and Philip 2008).
Aspergillus niger is a fungus, one of the most common species of the genus Aspergillus.The genus Aspergillus isimportant economically, ecologically and medically (Nizamuddinet al.2008).
Glucose oxidase enzymewas producedthrough the microbial fermentation. For that purpose solid state fermentation wasdevelopedwithA.niger. Solid state fermentation was applied to utilize agricultural residue such as Molasses as substrate.
The current research work was focused on production ofextracellular Glucose Oxidase (GOX) fromAspergillusniger using industrial waste such as molasses as substrate by Solid state ( static ) fermentation.
Availability: Items available for loan: UVAS Library [Call number: 2219-T] (1).
25.
Prevelance Of Brucellosis In Aborted Women Visiting Tertiary Care Hospitals Of Lahore City
by Saba Yasmin (2009-VA-211) | Prof. Dr. Aftab Ahmad Anjum | Dr. Tayyaba Ijaz (Co Supervisor) | Prof. Dr. Khushi Muhammad | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Pakistan is an agriculture based country whose rural population depends upon livestock for livelihood. Contribution of livestock to agriculture sector is 55.9 percent while 11.8 percent to the national GDP during 2013-14 (GOP 2013-2014). A number of infectious diseases hamper the growth of livestock sector. Some of the livestock diseases are zoonotic in nature and threat to human health. Brucellosis is considered among major zoonotic diseases throughout the world. The Mediterranean Basin, south and Central America, Eastern Europe, Asia, Africa, the Caribbean and the Middle East are considered as high-risk countries (Memish 2001).
Brucellosis in human beings is a major concern of community health. It causes acute and chronic illness, physical incapacity and loss of health. Bacterial species involved include Brucella abortus, Brucella melitensis or Brucella suis. Brucellosis is acquired by human beings from infected animals by close contact with vaginal secretions, urine, feces, blood, aborted fetus, or consumption of unpasteurized milk or other raw milk products. Shepherds, milkmen, butchers, knackers, veterinary assistants and abattoir workers are at high risk (Agasthya et al. 2007). Prevalence of brucellosis recorded by Mukhtar and Kokab (2008) in abattoir workers of Lahore Pakistan was 21.7 percent. Higher prevalence of brucellosis was observed in females (37.06%) than males (24.2%) in patients admitted at Peshawar, Pakistan (Shahid et al. 2014).
Symptoms of disease vary among human patients, ranging from non–specific, flu-like symptoms (acute form) to undulant fever (chronic form). Some of the serious complications of skeletal system, cardiovascular and central nervous systems may develop. Other important signs observed include arthritis, orchitis, epididymitis, abortion, retained placenta and stillbirth (Baba et al. 2001; Grilló et al. 2006). In animals, brucellosis in most of the cases results in abortion, birth of weak calves, death of young stock, infertility in males and reduced milk yield in females (Maadi et al. 2011; Abubakar et al. 2012).
There is actual need for teamwork between public health officials and veterinary officers to reduce communication of brucellosis between animals and human in endemic areas (Jelastopulu et al. 2008; Makis et al. 2008). Clinical picture of brucellosis is nonspecific and may vary from patient to patient. Therefore, laboratory diagnosis by isolation and culture or recognition of specific anti–Brucella antibodies is essential for confirmation of brucellosis (Al-Attas et al. 2000).
Diagnosis of brucellosis by culture and phenotypic description is time-consuming. Furthermore, risk of infection to worker is always there. Serological tests are commonly preferred for brucellosis in cattle and small ruminants, especially at farm level screening. Chance of cross-reactions with other gram negative bacteria is a major problem. Rose Bengal Plate Agglutination Test (RBPT) and Slow Agglutination Test (SAT) are extensively used for detection of anti-Brucella antibodies (Halling et al. 2005). Enzyme Linked Immunosorbent Assays (ELISA) have been developed to resolve suspected samples by RBPT. ELISA is more sensitive, so it can detect Brucella carriers which are negative by RBT, SAT and CFT (Aert et al. 1984). Molecular techniques are more reliable and specific than serological tests. Final confirmation of brucellosis is carried out using polymerase chain reaction (PCR), a molecular technique. Real-time PCR offers enhanced sensitivity, specificity and rapidity of performance when compared to conventional PCR (Gwida et al. 2012). Availability: Items available for loan: UVAS Library [Call number: 2225-T] (1).
26.
Polymorphisms Of Bovine Tumor Necrosis Factor Alpha Gene And Its Association With Mastitis In Sahiwal Cows
by Huma Sattar (2013-VA-03) | Dr. Sehrish Firyal | Dr. Ali Raza Awan | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Mastitis is one of the shocking maladies of milch animals causing high production losses to livestock industry of Pakistan (Kenyanjui et al. 2011). It is an inflammatory condition of udder; represent a major problem in dairy cow management. It is one of the most common and frequent disease of dairy industry. Producers suffer a huge loss due to veterinary treatment costs and necessary culling of the infected animals. It negatively affects the milk production, quality of milk, and farm economics (Fourichon et al. 2005). Increasing the disease resistance among dairy cattle is therefore desirable because without controlling mastitis, the national goals of developing dairy farming on commercial and scientific lines and production of wholesome milk which conforms to the standards of WTO Accord would remain elusive.
Mastitis is inflammation of udder that caused by physiological and metabolical changes (Schalm and Noorlander 1957). There are two main types of mastitis; clinical mastitis (characterized by classical symptoms i.e., swelling of udder, redness, clumps and clots in milk etc) and sub-clinical mastitis (not show any symptoms, Milk appear normal, udder appear normal) (Schrick et al. 2001). Mastitis is ranked as a top disease of dairy herds (Rinaldi et al. 2010). This mammary gland infection caused by pathogenic micro organisms such as Staphylococcus aureus, Streptococcus uberis, and Esherichia coli in the mammary gland (Heringstad et al. 2000).
India, China and United States are the larger producer of milk and Pakistan is on forth number in milk yield. Pakistan almost produces 36.5 million tons of milk yeild per year (Cady et al. 1983).The Sahiwal breed is well known among for its superior dairy qualities (Barker et al. 1998). Both cross and pure breed Sahiwal cows have high milk production rate (Khan et al. 2013).
It is very difficult to comprehend this disease because numerous environmental and genetic factors are involved in the origin and development of mastitis (Bradley 2002; Carvajal et al. 2013). Susceptibility and resistance to mastitis is a complex trait influenced by genetic variation of animals. Among these variations, the polymorphisms in immunity genes are principal key factors in defensive mechanism of mammary gland (Ibeagha-Awemu et al. 2008).
The mammary gland tissue is protected by immune system by two defense system; innate and acquired immunity. Innate immunity response by the host is a quick response of bacterial defense system (Mesquita et al. 2012). Innate system is a rapid and effective mechanism that activated on recognition of antigen (Akira et al. 2006). Innate immune system is activated when specific pattern recognition receptors (PRR) that are present on the surfaces which are attach to the specific pathogen (Shuster et al. 1996). PRR are presnt on leucocytes in milk and on the epithelial cells lining of udder. It is reported that T- lymphocyte subset i.e., CD4+, CD8+ and ɤδT are present in infected bovine mammary glands. (Goldammer et al. 2004; Strandberg et al. 2005).
Innate defense (nonspecific) of the mammary gland is stimulated by the physical barrier such as teat end, natural killer (NK) cells, neutrophils, macrophages and certain other soluble factors. The teat cannals are considering the main line of defense. Microorganisms enter from teat canal in milk. The main roles of teat sphincter muscles are to remain orifice close so that bacteria cannot enter. This teat canal also lined with keratin, whose estrified and non estified fatty acid function as bacteriostatics that provide protection and play role to eliminate bacteria causing mastitis (Oviedo-Boyso et al. 2007).
If a pathogen is not eliminated by the physical barrier, the acquired immune system is triggered. In comparison, this system is much faster than other immune response. The memory response is significantly stronger, long durable and more efficient to kill the pathogen. The acquired immune system (memory response) have ability to differentiate self or nonself cells and produce antibodies only against antigens through membrane bound protein called major histocompatibility complex (MHC) molecules. Specific immune system activate only when antigens bind with an MHC that is present on the surface of certain cells, this process is referred as antigen presentation. Recognition of pathogenic factors for elimination is mediated by macrophages, several lymphoid, and immunoglobulins (Ig) or antibodies (Sordillo and Streicher 2002).
The most acute responding macrophages and T-cell cytokines are TNF-α, LTF, IL1, IL6, IL8, and IFN-ɤ present in intramammary infection in cows. These genes play important role in improvement of immunity to mastitis (Burton and Erskine 2003).
Tumor necrosis factor alpha is main pro-inflammatory adipokine that is part of systematic immune defense. The main function of TNF-α gene is responsible for proliferation, differentiation and activity of many immune system cells; B lymphocytes, NK (natural killer). It also induces the production and release of many other cytokines (Wojdak Maksymiec et al. 2013) and also enhances the chemotactic and phagocytic effects of immune response. TNF-α gene contains four exons and three introns that are present on chromosome BTA23q22 (Bannerman 2009; Moyes et al. 2009).
TNF-α is a member of a group of cytokines that stimulate the specific immune system. TNF consist of 212 amino acid arranged in stable homotrimers (Kriegler et al. 1988; Tang et al. 1996). The 17-kilodalton (kDa) TNF protomers are composed of two β-pleated sheets and β-strands, joined together antiparallel (Tang et al. 1996).
TNF-α is a component of natural protection systems of humans and animals. Milk gives nourishment and disease resistance to the new born. Various cellular and soluble immune components are important for protecting the mammary gland from infectious diseases like mastitis. Mastitis affects one third of all dairy cows and cost the dairy industry about 2 million dollars annually (National Mastitis Council (1996). Dairy cattle are especially susceptible to mastitis due to diminished mammary gland defense mechanisms (Sordillo and Streicher 2002).
TNF-α is not only produced by activation of macrophages, but also other cell types such as CD4+ lymphocytes, NK cells, neutrophils, mast cells, eosinophils, and neurons. Large amounts of TNF are released in response to lipopolysaccharide, other bacterial products, and Interleukin-1 (IL-1).TNF-α stimulates the proliferation, differentiation and activity of many immune system cells; B lymphocytes, NK (natural killer). TNF-α induces the release of many other cytokines (Wojdak-Maksymiec and Mikolajczyk 2012). TNF-α also enhance the chemotactic and phagocytic effects of immune response.
. The present study is designed to determine the genetic polymorphism in exon 4 of TNF-α gene of mastitic cows and its association resistance and susceptibility towards mastitis.
Availability: Items available for loan: UVAS Library [Call number: 2224-T] (1).
27.
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).
28.
DNA Based Characterization of Xylanase Gene From Hyperthermophilic Archeon
by Saima Zulfiqar (2012-VA-539) | Dr. Muhammad Tayyab | Dr. Faiza Masood | Dr.Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Blank CD Availability: Items available for loan: UVAS Library [Call number: 2233-T] (1).
29.
Sequence Analysis Of Mitochondrial Atpase 8/6 Gene Variants In Equine
by Kashif Hameed Anjum (2012-VA-905) | Dr. Asif Nadeem | Mr.Maryam Javed | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Human has been using horses for doing different jobs like transportation, hunts, carrying loads, warfare and sports (Zhang et al. 2012). In Pakistan, horses and donkeys are mostly used for transportation whilehorses are also used for racing and playing games like polo.There are two main types of horses:Equuscaballusare domesticated horses and Equusferus are the wild horses. There are more than 300 breeds of horses in the world today (Barbara and Dafydd, 2007). The horse population is estimated as 0.32 million and has been decreasing over the years in Pakistan. Main breeds of horses that are found all over the Pakistan are Kajlan, Kakka, Balochi, Morna, Shien, Anmol, Makra, Pak-thoroughbred,Heerzaiand Waziri (Khan, 2004).
Seventy percent of the population earns living from the land. Agriculture contributes nearly 21% to gross domestic product and generates 43% of all jobs. Over 30 million people in rural areas derive their livelihood from livestock production. The number of impoverished communities moving from the country to find work in Pakistan’s towns and cities is rising. Many of these people rely on working equine animals to earn a living.
Nuclear and mitochondrial genomes are frequently used in animal genetic research. Nuclear genomeis generally a huge and complicated molecule and is not well studied in many species. However mitochondrial DNA being small sized and having high mutation rate is used frequently for the purpose of genetic research (Stanley et al. 1994). Characteristic of having fast evolution rate as compared to nuclear DNA makes mitochondrial genes a good tool for genetic studies (Avise, 1994).
Several studies have investigated the genetic relationship among horse and donkey breeds using mitochondrial sequences as a marker for breed characterization and phylogenetic. Each mitochondrion contains its own circular DNA, replication, transcription and translation machinery and serves as semi-autonomous organelle. Mitochondria perform so many important functions in our body like metabolism(oxidative phosphorylation), apoptosis and aging(Weinberg, 2007).
The advent ofpolymerase chain reaction and direct sequencing techniques with the use of mtDNA as a phylogenetic marker has been extended to much greater levels of phylogenetic inclusiveness (Zardoya and Meyer,1996). The special features of mtDNAi-e,lack of introns, maternal inheritance, absence of recombination events and haploidy have made it the most common type of sequence information used to estimate phylogenies among both closely and distantly related texa(Meyer, 1993).
Four of the five mitochondrial respiratory chain complexes, namely C1, C3, C4 and C5 (ATP synthase) contain subunits encoded by mitochondrial DNA (Kadenbach, 2012). ATP synthase (Complex5) functions to make ATP that is used by the cell (Von et al. 2009). ATP synthasecomprisesan integral membrane cylindrical, the F0 particle and a peripheral matrix-facing F1 particle, the catalytic ATP synthase domain (Boyer, 1997). All aerobically respiring organisms possess ATP synthase enzymes and are located inthe cell membrane in prokaryotes, the mitochondrial inner membrane in eukaryotes and the chloroplast thylakoid membrane (Ackerman and Tzagoloff, 2005). This enzyme is responsible for the final step of oxidative phosphorylation. The protons move down their concentration gradient from inter membrane space to matrix through F0 particle while F1particleuses the energy provided by influx of these protons and converts ADP molecule into ATP. ATPase 6 and ATPase 8 proteins are components of F0 particle where they play direct role in maintaining the structure and function of ATP synthase (complex 5). All five subunits of F1 and most of the F0 subunits are nuclear encoded(Collinson et al. 1996). Only two proteins i-e, ATPase 6 and ATPase 8 are encoded by mtDNA (Boyer, 1993).
The present study is designed to investigate the diversity and phylogenetic analysis of Thoroughbred Pakistani horse and donkey breeds on the basis of ATPase 6 and ATPase 8 genes.
Availability: Items available for loan: UVAS Library [Call number: 2236-T] (1).
30.
DNA Based Characterization Of Arginase Gene From Geobacillus Sp. SBS-4s
by Raabia Bibi (2012-VA-537) | Dr. Muhammad Tayyab | Dr. Abu Saeed Hashmi | Dr. Muhammad Imran.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Geobacillus is a group gram-positive, rod-shaped, aerobic, endospore-forming and obligate thermophilic bacteria, isolated from the diverse habitats, hot springs, thermal environments, terrestrial soils, deep sea sediments (Zeigler, 2014), petroleum and soil of desserts (Claus and Berkeley 1986). It grows at a wide range of temperature from 45 to 75°C and pH ranging from 6.2 to 7.8 (Nazina et al. 2001). These bacteria survives at higher temperature where most of other living species fail to survive (Claus and Berkeley 1986). Geobacillus have achieved a significant population with a worldwide distribution, probably in large part due to adaptive features of their spores (Zeigler, 2014). These can be found singly or in short chains and motile by means of peritrichous flagella and is capable of secreting a wide variety of extracellular and intracellular enzymes i.e amylase, lipase, carboxypeptidase, cellulase, xylanase, protease and galactosidase (Fogarth et al. 1974; Obeidat et al. 2012).
Geobacillus sp. SBS-4S was isolated from hot spring located in Gilgit, Northern areas of Pakistan. It was found to be an aerobic, gram-positive and rod-shaped bacteria having ability to hydrolyze a variety of sugars, carboxylic acids and hydrocarbons at elevated temperatures from 45 to 75°C. SBS-4S was found to be involved in the production of various intra and extra cellular enzymes (Tayyab et al. 2011).
Arginase is the enzyme responsible for the degradation of arginine resulting in the production of urea and ornithine (Kaur et al. 2009). It is accomplished by the cleaving of the guanidinium group from arginine which yields urea (Turras et al. 2008). Arginase present in many mammals (Homo sapiens), Bacilli (cyanobacteria), protozoa (Entamoeba histolytica), yeast (Saccharomyces cerevisiae), fungi (Neurospora crassa) and plants (Lathyrus sativus) etc (Kaur et al. 2009). The crystal structure of arginases have been determined by X ray crystallographic studies. This is a manganese dependent enzyme. The enzyme shows its activity through the metal ion. Metal ion is actively responsible for the incorporation of water molecules essential for the activity of the enzyme. A second proposed mechanism, based on electron paramagnetic resonance (EPR) studies postulates direct coordination of the substrate to manganese and disruption of the aqua bridge. Arginases are homo-oligomers, with a typical subunit mass of 32 to 36 kDa (Bewley et al. 1999).
There are two types of arginases, arginase-I and arginase-II, located in the cytoplasm and mitochondria, respectively. The principal ureagenic enzyme activity arginase-I is most abundant in normal mammalian liver and acts in coordination with the other enzymes of the urea cycle to sequester and eliminate excess nitrogen from the body. The second form arginase-II can be found in many organs, with the highest levels found in kidney and prostate where as lower levels in macrophages and lactating mammary glands (Iyer et al. 2002).
Important role of arginase in controlling the cellular levels of arginine and ornithine, which are required for various critical metabolic processes, including protein synthesis and the production of creatine, polyamines, proline and nitric oxide (NO). Type II arginase is found in a variety of different tissues and have a key role in the regulation of urea cycle arginine metabolism by regulating levels of arginine in the cell (Bewley et al. 1999). The enzyme arginase plays key role in the pathogenesis of pulmonary disorders such as asthma through dysregulation of L-arginine metabolism and modulation of nitric oxide (NO) homeostasis and it also play role in the development of chronic airway remodeling through formation of ornithine with downstream production of polyamines and L-proline, which are involved in processes of cellular proliferation and collagen deposition (Benson et al. 2011). Arginase involved in tissue repair processes by the synthesis of L-ornithine, which is the precursor of polyamines and proline that are involved in cell proliferation and collagen synthesis (Maarsingh et al. 2009).
Genetically engineered arginase as fusion protein with prolonged half-life and increased efficacy are used to treat different tumor lines that inhibit cell proliferation and impaired cellular migration in vitro and in vivo (Li et al. 2013). This is a arginine-degrading and ornithine producing enzyme and is used to treat arginine-dependent cancers (Yu et al. 2013). Chemically modified arginase-II has been employed for the treatment of taper liver tumor and L5178Y murine leukemia (Kaur et al. 2009). The enzyme was cloned and expressed in E. coli and subsequently conjugated to polyethylene glycol to increase the circulating half-life and decrease the immunogenicity of the recombinant mycoplasma enzyme. The human hepatocellular carcinoma, melanoma cell lines and tissue samples do not express argininosuccinate synthetase (ASS), making them auxotrophic for arginine and thus reasonable candidates for arginine deprivation (Yang et al. 2010).
Arginase is induced in murine myeloid cells mainly by T-helper 2 cells cytokines and inflammatory agents and participates in a variety of inflammatory diseases by down-regulation of nitric oxide synthesis, induction of fibrosis and tissue regeneration. In humans, arginase I is constitutively expressed in polymorphonuclear neutrophils and is liberated during inflammation. Myeloid cell arginase-mediated L-arginine depletion profoundly suppresses T cell immune responses and this is a fundamental mechanism of inflammation-associated immunosuppression. Pharmacological interference with L-arginine metabolism is a novel promising strategy in the treatment of cancer, autoimmunity or unwanted immune deviation (Munder, 2009).
Arginase has very important role in nitrogen fixation and fruit ripening (Yu et al. 2013). Putrescine (1,4-butanediamine) is the product obtained from arginine with the highest market value and it is used as an intermediate in a large number of industries, including the pharmaceutical industry, agrochemical industry and textile industry (Turras et al. 2008).
Arginine is a semi-essential amino acid and is the precursor for the formation of nitric oxide (NO) by nitric oxide synthases (Getz and Reardon, 2006). One of the major functions of arginine within the body is as an intermediate in the urea cycle. In the cytosol of hepatocytes, arginase-I removes the guanidine group from arginine to produce urea and ornithine. Urea is then transported from the hepatocyte into the bloodstream and ornithine is used to regenerate arginine within the hepatocyte. Arginine deficiency causes several disorder like, hyper cholesterolemia, hypertension, diabetes mellitus, kidney failure, hyper homo-cysteinemia, smoking, and aging (Alvares et al. 2012). Arginine is used to modulate the cellular immune response during infection. The generation of nitric oxide from arginine is responsible for efficient immune response (Das et al. 2010).
Arginine is synthesised in humans and other mammals from citrulline in two steps through the urea cycle enzymes, argininosuccinate synthetase (ASS) and argininosuccinate lyase (ASL). ASS catalyses the conversion of citrulline and aspartic acid to argininosuccinate, which is then converted to arginine and fumaric acid by ASL (Yang et al. 2010).
Ararinase play important role in conversion of arginine to 1,4–butanediamine (a building block for nylon-4,6), through two main transformations: the hydrolysis of arginine to ornithine and urea; and the decarboxylation of ornithine to 1,4–butanediamine and carbon dioxide. Both steps can be catalyzed chemically or enzymatically (Turras et al. 2008).
The present study deals with the characterization of arginase gene.
Availability: Items available for loan: UVAS Library [Call number: 2244-T] (1).
31.
Application Of Microsatellite Markers For Genetic Diversity Analysis Of Endangered Punjab Urial (Ovis Orientalis Punjabiensis) In Pakistan
by Anam Aftab (2012-VA-534) | Dr.Tanveer Hussian | Dr. Wasim Shehzad | Dr. Muhammad Tayyab .
Material type: ; Literary form:
not fiction
Publisher: 2014Dissertation note: Biological diversity is now recognized as common concern of mankind and genetic diversity is the major driver of variation within and across breeds, which helps populations to adapt to environmental changes. There is very little importance is given to conserve wild sheep in previous years and its genetic diversity is decreasing day by day. Every now and then breeds are being haunted for crossing to some other imported breed without attempting to see if such efforts will be sustainable. For any breed development efforts thus, available genetic resources need to be characterized both at phenotypic and genetic levels (Khan et al. 2007).
Among the three levels of Biodiversity, one is Genetic variation which is suggested bythe International Union forNatureconservation (IUCN) for preservation (Mc Neely et al. 1990). The reason for it is that firstly; genetic diversity favors the changes as the environment changes and secondly; it prevents inbreeding depression (Reed and Frankham 2003). In this way genetic diversity increase the survival status and increase fitness of individuals.
Among many other wild animals present in Pakistan, there are 6 to 9 species of wild sheep (Ovis orientalis) are present which have different color and size of their winter neck ruff of males, saddle patches and horns color. Urial is a picture of Marco Polo in texture and hue. In Pakistan, ladakh urial, Blanford urialand Punjab urial are found in Gilget, Baluchistan and Punjab respectively. The discrepancy lies in the color of ruff among these 3 sub species (Roberts, 1977). Urial is among those precious wild animals that were hunted severely for trophy and other purposes in the past, that’s why included in red list of IUCN in vulnerable category (IUCN 2000).
Punjab Urial (A type of wild sheep – Ovis vignei punjabiensis) belongs to family bovidae which is the large family consisting of 140 species (Glazko et al. 2011) is facing serious threat of extinction in Pakistan is a medium-sized wild sheep which is included in IUCN red list of endangered animals (IUCN 2002). Urial is inhabitant in Western Central Asianregion stretching from northeast side of Iran and west side of Kazakhstan to Balochistan (Pakistan) and Ladakh regions of North India. The local name of Urial is Shapo, Arkar and Gad. Reddish-brown outstretched pelt that achromatizes during the winter is one of the distinct traits of urial (Aleem 1977; Schaller 1977). Urial is gregarious and sexually dimorphic as males are called rams and females as eves (Awan 2001). Male have weightof 40 kgand have large spiralled horns having height of 80 to 100 cm and females have comparatively less weight and height of 25 kg and 12 cm respectively and have uncurled horns.Females give birth to 1 or 2lambs in recent days of April (Awan, 2001). Males have a black ruff expanded from the neck to the trunk and notably longer horns. Table 1.1:Some physical features of the Punjab Urial (Awan et al. 2001)
Features Urial
Body weight 40 kg male; 25 kg female
Shoulder size 31-35 inches or 80-90 cm
Horn size 39 inches or 80-100 cm long male; 12 cm long female
Urial are found in moderate to very arid habitats, especially grasslands including agricultural fields and woodland areas (Valdez, 1982). Urial is herbivorous and eats grasses, shrubs and grains. The patch of salt range of Pakistan which fall in the area of Pind Dadan Khan, Choa Saidan Shah and Kallar Kahar is considered like a paradise on earth for the wild fauna. The fascinating hills of these areas are covered with thick trees and different wild plants are also the sanctuary of urial.
Table 1.2: The details of these sub species in IUCN list of endangered mammals (IUCN 2002).
Subspecies Citation in IUCN list
Ovis vegnei blanfordi VUC 1 Appendix ll
Ovis vegnei punjabienses ENA1cde,c1+2a Appendix ll
Ovis vegnei vegnei VUC 1 Appendix ll
In Pakistan, Punjab Urial dispersed throughout the Kala Chitta and Salt Range in a very little number(Hess et al, 1997). The Afghan Urial inhabits Baluchistan, Khyber Pakhtunkhwa, and Sindh Provinces. In Chitral District, little segregated populations of Ladakh urial are stillextensively distributed near the west bank of the Kunar river from Chitral southwards to Drosh. Ladakh Urial existence at the east bank of Kunar river and north of Chitral are not proved (Malik 1987). Total population estimate of urial is recorded by conducting several surveys. Reasonably 2,500 to 3,000 urial existsin Baluchistan (Hess et al. 1997). In 1993 the overall population assessment of Northern Areas was four hundred to five hundred Urial (Rasool 1999). In Pakistan, there were seemingly less than 600 Ladakh urial (Hess et al. 1997;NWFP 1992; Schaller 1971, 1977) but the number of urial decreased to 200 to 300 urial in all over northern areas (Rasool 1999). Based on the facts mentioned above there is dire need to conserve the Urial population in the country. The urial is one the precious fauna of Pakistan and provides us with wool and meat (fat, flesh or any eatable part). It is also important in economic way and in maintaining of ecosystem balance. In the beginning, sheep were reared for meat, milk and skin (Ensminger and parker, 1986). After 3500 B.C. men learnt to spin wool and so used wool in textile industry (Smith et al. 1997). So, because of increasing world population, there is great demand of these products and increasing day by day. That’s why we have to conserve is natural resource of Pakistan.Habitat fragmentationleads to the risk of exaggerated genetic drift and inbreeding in isolated population. So, there is need to save urial from these threats by enforcement of law and conserving it for future.
In all over the euchromatic genome Microsatellite markers are present. These markers are highly polymorphic (Ellegren 2000; Schlotterer 2000).A lots of polymorphic microsatellites have been analyzed in ruminants like domesticsheep, cattle etc. (de Gortari et al. 1997,1998 ;Hayes et al.1996; Jenkings et al. 1997) aiding the use of these in parentage testing.
Microsatellite markers are among the most reliable molecular markers for genetic characterization studies in animal species (Sunnucks, 2001) and are simple sequence repeats (SSRs) of 1-6 base pairs, repeated tandemly in coding as well as noncoding portion of DNA in prokaryotes and eukaryotes (Weber and May, 1989; Toth et al., 2000). Microsatellite markers have often used for genetic diversity studies because they areabundant, unbiased, widely distribution all over the DNA, highly polymorphic, easy in assessment like genotyping of these markers (Canon etal. 2001).Microsatellite markers aid in genetic differentiation and conservation studies (Peter et al. 2007;Rendo et al. 2004; Arranz et al. 2001).These are considered very useful markers for assessment of genetic diversity, parentage confirmation, genome mapping, disease research population genetic studies and conservation genetics. These are also reported to be efficient enough to identify within and among breed differentiation and population sub structuring in cattle (Glowatzki-Mullis et al. 1995; Ciampolini et al. 1995; Garcia-Moreno et al. 1996; Jarne and Lagoda, 1996; MacHugh et al. 1998).Therefore the conservation activities are very important to save Punjab Urial from extinction and the study is designed to explore its genetic diversity.
Availability: Items available for loan: UVAS Library [Call number: 2261-T] (1).
32.
Detoxification Of Aflatoxins Using Different Organic Acids
by Sana Ejaz (2013-VA-14) | Dr. Mateen Abbas | Dr. Muhammad Tayyab | Dr. Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: From global prospective of food safety and food security, mycotoxin contamination of foods has gained much attention as potential health hazards for humans and animals. Cereals and other crops are exposed to fungal attack in the field or during storage and this attack may result in mycotoxin contamination of crops. Animal feed is basic necessity for all the live stock, poultry and other animals. AF is the most important for human and animal health perspective and in developing countries such as Pakistan where climate conditions favor the formation of these toxic metabolites. Governments and private organizations of international level have established maximum residue levels (MRIs) which usually guide to control AF in feed. Therefore, the current study was planned to detoxify AF by using different organic acid treatments in animal feed collected from different dairy farms of Punjab.
The samples of cotton seed cake, maize oil cake and animal feed were collected and checked the presence of AFB1 qualitatively by TLC and quantitatively by HPLC. The samples which gave positive results were treated with different acidic treatments applied on it. Firstly checked the results of citric acid, acetic acid and lactic acid on feed sample qualitatively by TLC. TLC plates were checked under UV box and the samples which showed the detoxification of AF were quantitatively analyzed by HPLC in Toxicology Laboratory, QOL, UVAS, Lahore, Pakistan.
The average concentration of AFB1 found in the cotton seed cake, maize oil cake and mixed feed were 279.8 ppb, 34.2 ppb and 25.5 ppb, respectively much greater than permissible levels proposed by European Union. Treatments of varying concentration of citric acid, acetic acid and lactic acid were applied on positive samples (≥20 ppb) and checked their effect on rate of detoxification.
All the above mention treatments applied on the feed samples in order to obtained in vitro detoxification of AFB1. Sprayed different concentration of acetic acid, citric acid and lactic on positive samples by varying volumes and placed them over night then extracted and analyzed.
It has been observed that 1N concentration of citric acid, acetic acid and lactic acid showed complete detoxification. However, when these samples were treated with 0.5N solution of organic acids then variation was seen in rate of detoxification.
Statistically these results were analyzed by ANOVA which showed that effect of these treatments on rate of detoxification was highly significant (P<0.05). In vitro detoxification of AF by these organic acids was proved beneficial in order to reduce the animal and human health risks. However, in vivo detoxification of aflatoxin by using these organic acids should be studied in future.
Availability: Items available for loan: UVAS Library [Call number: 2283-T] (1).
33.
Genetic Association Study Of Apolipoprotein A-V (Apoa5) And Sortilin (Sort1) Genes With Risk Of Coronary Artery Disease
by Irfan Basharat (2012-VA-802) | Dr. Akhtar Ali | Dr. Wasim Shehzad | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: In developed countries cardiovascular disorders are prominent cause of death. One third deaths in the world are due to cardiovascular disorders. Among cardiovascular disorders coronary artery disease responsible for one in five deaths in USA. Its main reason is the lipids values particularly cholesterol and triglycerides in the blood. An estimation made by WHO indicated that 9 million people die per year due to hypercholesterolemia.
100 blood samples were collected from patients of coronary artery disease and from normal patients with no myocardial history. Allele specific primers for SORT1 gene and APOA5 genes were designed using Primer 3 software web facility. Genomic DNA will be amplified by PCR then genotyping will be carried out and DNA will also be sequenced.
Hardy-Weinberg principle and Fisher Exact test were used to assess the allele frequency and significant variations from results
When patient of MI and normal group were genotyped and sequenced we find out that there are 34 AA homozygous, 1 GG homozygous and 12 heterozygous persons in case of APOA5. The SORT1 person shows 24 GG homozygous and 3 AA homozygous and 13 heterozygous persons.
Our study shows a definite association between APOA5 and SORT1 with respect to MI disease persons. This study shows a significant association of single nucleotide polymorphism in APOA5 and SORT1 genes with coronary artery disease. Availability: Items available for loan: UVAS Library [Call number: 2326-T] (1).
34.
Variation Analysis Of Hepatitis C Virus Gene Encoding E2 Glycoprotein
by Saimoon Theeen (2009-VA-565) | Dr. Muhammad Imran | Dr. Sehrish Firyal | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Hepatitis C virus (HCV) major cause of liver infections was discovered in 1989. It is positive stranded RNA virus and belongs to Flaviviridae family. Its genome shows high rate of variations due to which, its rate of infection is high. As in Pakistan 3% to 6% population and 170 million people worldwide are affected by it. Due to variations in its genome it is classified into 6 major genotypes and these genotypes are further classified into many subtypes. Size of HCV is about 9500 bps which only encodes single polyprotein. This 3000 to 3300 amino acids polyprotein is processed by cellular and viral proteases to generate 10 polypeptides consisting of 4 structural (Core, E1, E2 and P7) and 6 non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A, NS5B). Whereas E2 is considered the most immunogenic gene from all the genes. It involves in the interaction with the host cell and easily escape from the immune system of host due to the presence of hypervariable regions in E2 gene.
To isolate the E2 gene RNA extraction was done using the kit method and then it was converted to the cDNA which is then followed by the PCR amplification. The amplification products were then purify and sent for the sequencing to CAMB. Then the bioinformatics tools were applied on the results. In which the protein structural analysis and epitope mapping was done. Then the conserved epitopes were predicted using the IEDB conservancy analysis tool. The conserved B-cell epitopes (TElAILPCSFTPMPAL and RGERCDIEDRSEQH) and T-cell epitope (TPMPALSTG) are now considered valuable to produce the antibodies against E2 protein.
For diagnosing HCV genotype 3a, these conserved epitopes may be highly useful and may also help in developing a successful vaccine that can target 3a genotypes. Availability: Items available for loan: UVAS Library [Call number: 2333-T] (1).
35.
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).
36.
Genetic Effect Of Cholesteryl Ester Transfer Protein (Cetp) Gene In Coronary Heart Disease Patients
by Zakiya Bano (2013-VA-554) | Dr. Akhtar Ali | Dr. Waseem Shehzad | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Cholesteryl ester transfer protein (CETP) gene takes part with certain reverse cholesterol
transport (RCT) pathway for the excess amount of accumulated lipid in peripheral tissues. The
variations in this gene due to missense mutations on different exonic, intronic or on promoter
regions alter CETP activity as well as impair the RCT pathway. By which, lipid metabolism also
effects and causes atherosclerosis in vessels which trigger the blockage of blood flow and induces
the imbalance for the supply of oxygen to the heart. So this atherosclerosis directly involves in
addition of risk factor for coronary heart disease. Preferable study was made to highlight effect of
CETP gene at molecular level by comparing control group with the selected patients having
coronary heart disease. This study was appreciably made possible by targeting two reported
polymorphisms, one in the intron 1 region Taq IB (rs708272) and on exon 14 region I405V
(rs5882) of this CETP gene. The study was relatively speculated by the extraction of genomic
DNA from all selected blood samples. By selecting two primers, certain segments were amplified
for both rs708272 and rs5882 polymorphisms. Analysis of allelic frequencies distribution was
calculated by Hardy Weinberg Equilibrium which showed no significance among control and
CHD group and there was no association was analyzed in our population by using Fisher’s Exact
Test. This is because of small number of samples studied in our population. But maximize
concentrations of lipid parameters such as TC, LDL and TG with minimum variation in HDL-C
concentration in CHD group as compared to control group that showed the effect of these
polymorphisms on the activity of CETP gene with coronary heart disease. These determined
missense mutations in CETP gene was helpful molecular tool for the screening purpose in coronary
heart disease patients. Availability: Items available for loan: UVAS Library [Call number: 2345-T] (1).
37.
Production Of Single Cell Protein By Using Banana Peels As Substrate And Its Biological Evaluation In Broiler Chicks
by Muhammad Sheraz Yasin (2012-VA-603) | Miss Shagufta Saeed | Dr. Muhammad Tayyab | Prof. Dr. Aftab Ahmed Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: The term single cell protein (SCP) refers to dead, dry microbial cells or total proteins extracted from pure microbial cell culture and is produced using a number of different microorganisms including bacterium, fungus and algae. It can also be called biomass, bioprotein or microbial protein.
Besides high protein content (about 60-82% of dry cell weight), SCP also contains fats, carbohydrates, nucleic acids, vitamins and minerals.
Fermentation media containing grinded banana peel as substrate was used to check the production of single cell protein for the selected Arachniotus sp. Different parameters were optimized for higher production of SCP e.g: Incubation period, pH, volume of inoculum, carbohydrate source, concentration of corn steep liquor and ionic salts concentration.
The biomass yield was estimated for total protein content by Lowrymethod. Biomass produced from fermentation was used for biological evaluation in feed trials of broiler chicks.
It is found that Arachniotus sp gave maximum single cell protein 7.49 g/L using 10 g banana peels at 72 hours incubation period. And protein concentration increased 7.58 g/L by optimizing volume of inoculum 2ml. It is observed in present study carbohydrate source also increases the protein concentration 8.41 g/L when carbohydrate source was optimized (glucose 3%).
Later on it was found that nitrogen source also enhance the protein production upto 12.61 g/L by using 2% corn steep liquor. Results also revealed that ionic salt concentration also play important role in the production of biomass protein, addition of 0.075% CaCl2.H2O produced 14.45 g/L single cell protein using above mentioned optimized conditions. 0.050 %
K2HPO4 produced 15.06 g/L. Addition of 0.050% MgSO4.7H2O produced maximum protein 15.86 g/L.
Biological evaluation in broiler chicks of this biomass protein shown there is no deleterious effects on weight gain, feed conversion ratio, protein efficiency ratio and net protein utilization. Maximum weight gain observed 215.6 grams in the group (C) in which 50% sunflower meal was replaced with biomass protein.
Feed conversion ratio in group (C) was 2.64 in which 50% sunflower meal was replaced by biomass protein and in group (B) was 2.51 in which 25% sunflower meal was replaced. And in control group (A) feed conversion ratio was 2.41.
Protein efficiency ratio was observed with non-significant value. And same results were shown by Chaves et al (1988) who reported non-significant differences among the standard and test diet when Chaetominumcellulolyticum biomass was fed to chicks. Net protein utilization observed in present study gave significant P value among the groups.
So it is concluded that single cell protein produced by this method is cheap and can be used in the food industry as food supplements and can also be included in poultry feed. The study findings suggested that microbial biomass produced by Arachniotus sp using banana peels as substrate can be replaced upto 50% of the protein supply by sunflower meal without any deleterious effects on growing broiler chicks. Moreover, it will also help in the reduction of pollution by using waste i.e. banana peel for useful purpose.
Availability: Items available for loan: UVAS Library [Call number: 2347-T] (1).
38.
Assessment Of Evolutionary Rate In Different Serotypes Of Foot & Mouth Disease Virus
by Muhammad Farooq (2011-va-823) | Dr. Ali Raza Awan | Prof. Dr. Thair Yaqub | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: FMDV belongs to the family Picornaviridae with seven serotypes around the world. Nevertheless, serotypes prevalent in Asia includes A, O and Asia-1. Because of evolution in genomic sequence of FMDV, it is becoming difficult to control the problem through conventional methods. Changes in the genome can be detected using software through sequence analysis. In the software, evolutionary models are used to measure the evolutionary change for the identification of new sub lineages.
In current study genomes sequence data (1998 - 2011) of bovine FMD serotypes (A, O and Asia 1) was collected through NCBI in FASTA format. This data was converted into PHYLIP format. On Dell workstation, with Microsoft Windows 8.1 operating system, BioEdit, TipDate V.1.2 was deployed. Sequence data was aligned through CLUSTAL W algorithm of Multiple Sequence Alignment using BioEdit. Using TipDate, genome sequence data was analyzed using three evolutionary models (F84, HKY and REV) and phylogenetic trees were produced showing evolutionary rate and likelihood ratio of FMDV serotypes O, A and Asia-1..
Results of the current study showed higher values of evolutionary rate in bovine FMD virus which was estimated 7.49 x 10-4 with likelihood value -1429.507680 in serotype A, 2.418 x 10-3 with likelihood -3707.168484 in serotype O and2.16 x 10-3 with likelihood value -3723.344884 in serotype Asia-1, respectively. Markove Reversible Model showed higher rates of evolution in all three serotype with best likelihood values. Phylogenetic results showed higher rate of evolution or substitution in viruses. Furthermore serotypes A, O and Asia-1are mutating with passage of time and new variants are being observed. It was also observed that this evolutionary process is continued in these three serotypes during 1998-2011.
This study confirmed the evolutionary changes in FMDV serotypes prevalent in Pakistan during the period 1998 – 2011. This study showed that isolate are evolving with increasing rate. High rate of mutation in Asia-1 was observed than serotype A and serotype O. F84 and HKY85 models produced close results
but these models are not identical works on equal and unequal base frequencies. Markove model estimated average base substitution with mutation and depicts good phylogenetic trees of sequence data. Availability: Items available for loan: UVAS Library [Call number: 2372-T] (1).
39.
Production Oflaccase From White Rot Fungususing Rice Bran As A Substrate By Solidstate Fermentation
by Muhammad Tanweer Muneer (2013-VA-06) | Mr.Shahid Abbas | Dr. Muhammad Tayyab | Prof. Dr.TahirYaqub.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Laccase are copper oxidases and are found in large quantities in several white rot fungi that are involved in lignin metabolism. Fungal laccases have boundless biotechnological functions across the globe like the decolouration and detoxification of industrial effluent, bleaching of pulp, phenolic elimination from wines, in preparation of biosensors in detergents blocking dye transfer- functions. Laccase showed vast variety of substrates due to this ability they can enhance different types of industrial mechanism such as methylation, demethylation, polymerization, mineralization of pollutants like hydrocarbons. White rot fungus is efficient for the production of laccase using agro-waste as substrate.
In this research white rot fungus was isolated from stock cultures of Department of Microbiology, University of Veterinary and Animal Sciences, Lahore, Pakistan and the organism was maintained on Tien& Kirk media slants and petri plates. Solid state fermentation technique was used using basal fermentation medium and agro waste rice bran was used as substrate for the production of laccase.Proximate analysis was performed of the substrate rice bran to analyse crude protein, fat, ash, moisture and fibre content. The fermentation was performed at room temperature and flasks were placed on orbital shaker at 100 rpm and 30˚C.
Enzyme activity was checked using (ABTS) as substrate at 420nm, for every 24 hour to observe the maximum enzyme production. Fermentation parameters like substrate concentration, incubation period, pH, temperature and nitrogen source (corn steep liquor and ammonium sulphate) were optimized. The concentration of substrate optimized for rice bran was 7.5g/100ml and optimum production of 6.11 IU/ml of enzyme was observed. The optimum day for the production of enzyme was day 7 and the amount of enzyme produced was 6.91 IU/ml. The optimum pH and temperature were 4 and 40˚C respectively, and the amounts of enzyme produced were 7.48 IU/ml and 7.96 IU/ml respectively. Two nitrogen sources optimized were maize steep liquor 1ml and ammonium sulphate 0.2 g, and the enzyme produced was recorded 7.67 IU/ml and 9.41 IU/ml respectively. Large scale fermentation batch of one litter was carried out under the optimized conditions and the enzyme produced was 9730 IU/L. Triplicates of each parameter were prepared.
The enzyme was purified using the purification techniques like ammonium sulphate precipitation, then by dialysis excess salt was removed, and then gel filtration was performed to collect different fractions on the basis of size of molecules and molecular mass of the laccase was analysed by SDS-PAGE. The size of the protein was found to be 70kDa. Characterization of laccase was performed in terms of optimum pH, temperature and in response to inducers and inhibitors. The optimum pH and temperature of the purified enzyme was 6 and 40˚C respectively. The inducer copper sulphate enhanced the activity of enzyme up to 9.7 U/ml and then inhibitors EDTA and 2-merceptoethanol reduced the activity level up to 4.17IU/ml and 3.98 IU/ml respectively. To study and analyse the effects of optimization parameters Pearson correlation, descriptive statistics and one way Anova were used.
The optimum production of laccase was achieved using agro waste rice bran. The enzyme produced was economical and it can provide effective solutions for bioremediation of hazardous compounds and pollutants.
Availability: Items available for loan: UVAS Library [Call number: 2377-T] (1).
40.
Genetic Identification And Molecular Classification Of Sub-Family Phasianinae Of Pakistani Bird Species Through Dna Barcoding
by Maryem Hussain (2008-VA-349) | Dr. Ali Raza Awan | Dr. Sehrish Firyal | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: DNA barcoding is a precise technique that uses molecular genetics tools for accurate identification, categorizing, relating and separating the phylogenies of species. Being a small sized genome and agile enough to show rapid mutation, mtDNA has been used as a pertinent marker of molecular biodiversity.The aim of this study was to develop DNA barcode for genetic characterization and classification of Sub-family Phasianinaeof Pakistani bird species. Theyhave not been genetically identified yet in Pakistan. It includes birds like domestic chicken(Gallus gallusdomesticus), aseel chicken(Gallus gallusdomesticus strain),blue peafowl(Pavo cristatus), green peafowl (Pavo muticus), white peafowl (Pavo cristatus leuticus), Kalij pheasant (Lophura leucomelanos),monal pheasant (Lophophorus impejanus),koklass pheasant(Purcrasia macrolopha), ring necked pheasant (Phasianus colchicus), Tragopan (Tragopan melanocepals) andred junglefowl (Gallus gallus). These birds are considered an important part of an ecosystembecause they play a significant role in seed dissemination, pollination of plants and disease spread which are the basic constituents of an ecosystem. They are used for food, hunting and entertainment purposes.
Mitochondrial geneCytochrome c oxidase subunit 1 (CO1)of 500bps was used as a marker for identification at specie level.Genomic DNA was extracted by each blood and tissue sample of eleven bird species (33 samples). Amplification of CO1 gene was a done by using a universal set of primers (BIRDF1 and BIRDR1)containing region of almost 750 bps (Hebert et al. 2003).Amplicons were purified and sequenced Sanger sequencing method (Sanger et al. 1977). Forward and reverse sequences were analyzed using softwaresEMBOSS merger,ClustalW, BioEdit and nBLAST. Phylogenetic analysis of selected bird species was done. Each sequence was aligned
with its reference sequences of CO1 gene present on NCBI. Every nucleotide position which did not align with the reference sequence was studied to identify SNPs. Fixation index (FST) were used to measure species diversity within a same sub population relative to that found in the entire population. Consensus sequences (500bps) generated was used to construct their phylogenetic tree to see their evolutionary relationship with other bird species. All species showed their closest linkage with their respective species. Pakistani population of peafowl and chicken species showed the close relation with same sequences generated in China. Tranopans showed its closest linkage with T. temminckii.
In conclusion, seven species ofPhasianinaesub-family of Pakistani bird species was genetically characterized first time in Pakistan by using CO1 as a barcode. It proves that DNA barcoding is an efficient and accurate molecular tool for species identifica¬tion and phylogenetic implication. This study leads to establish a DNA Data Bank that helped scientists to investigate the biodiversity, taxonomic classification, specie identification, in forensic purposes and to study the genetic and phenotypic evolution of these species. DNA barcoding through CO1 gene works as a functional tool for detectingmeat mislabeling and preventing illegitimate trade. This study has established foundations for molecular biologists to study taxonomic uncertainties at sub species level using SNP based identifying marker. 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: 2376-T] (1).
41.
Molecular Characterization of Pakistani Common Leopard
by Muhammad Usman Ijaz (2012-VA-908) | Dr. Sehrish Firyal | Dr. Ali Raza Awan | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: CD not available. Availability: Items available for loan: UVAS Library [Call number: 2379-T] (1).
42.
Genetic Effect Of Interferon Gamma On Bovine Resistance Against Mycobecterium Bovis
by Syed Ahmed Raza Rizvi (2012-VA-819) | Dr. Maryam Javed | Dr. Tanveer Hussain | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Bovine tuberculosis is a disease caused by the species included in the Mycobacterium tuberculosis complex. IFN-GAMMA are a family of conserved innate immune recognition receptors that trigger adaptive immune responses. IFN-GAMMA 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 IFN-GAMMA 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 IFN-GAMMA 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 IFNG mentioned in table. The Eight SNPs identified in the coding region of INTERFERON GAMMA 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, and P8 C >T. The one SNP found in the current research is in compliance with the (Sun et al. 2012) research on INTERFERON GAMMA hence Nine SNPs found in the current research are novel in Nili Ravi buffalo. Research on IFN-GAMMA hence Seven 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, among them four were transitions and four were transversion .
Population genetic analysis and allelic distribution at all loci was analyzed using
Summary
57
POPGENE 32 software indicated that at [P3=0.354539>0.05] , [P5=0.365524>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= 0.000032< 0.05], [P2=0.038766< 0.05] and [P7=000394< 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: 2419-T] (1).
43.
Lactoferrin Gene Polymorphism in Dairy Cattle
by Syeda Iqra Aiman Bukhari (2009-VA-556) | Dr. Sehrish Firyal | Dr. Ali Raza Awan | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Several factors militate against realizing the milk production potential of cows. Mastitis is the most costly and the prevalent production-limiting disease of dairy animals in Pakistan and elsewhere in the world. It is accompanied by elevated Somatic cell count (SCC) in the milk and estimated genetic correlation between SCC and mastitis ranges between 0.53-0.77. Susceptibility and resistance to mastitis is a complex trait and influenced by genetic variation of animals. Among these variations, the polymorphism in Lactoferrin gene (LTF) plays an important role in the immune response to mastitis.
Polymorphism in intron 6 of LTF gene is associated with mastitis susceptibility and resistance. It is a potential candidate gene for imparting resistance mastitis in dairy cows.
The present study was designed for the identification of polymorphism in LTF gene associated with mastitis. Milk and blood samples were collected from 20 Sahiwal cows having clinical and subclinical mastitis. SCC of milk samples was performed using serial dilutions. 10 normal Sahiwal cows as control were included in present study. DNA was extracted from blood using organic extraction and kit method followed by DNA quantification. Amplification of LTF gene was designed by using already reported primers obtained from NCBI.
LTF gene was amplified and sequenced to get the full length sequence of this gene. Comparative analysis of the resulted sequences using NCBI BLAST was done.
Outcomes:
The results obtained from polymorphisms in LTF gene can play an important role for selection of mastitis resistant and susceptible dairy cows. This can be useful in selective breeding of cattle for enhanced immune response, as a tool to improve inherent animal health, which ultimately can lay the foundations to contain the magnitude of economic loss due to mastitis.
Develop a biological response modifier that will promote a sustained immunity of the mammary teat and protect the gland from invading pathogens. Availability: Items available for loan: UVAS Library [Call number: 2416-T] (1).
44.
Physical, Chemical and Biological Treatment of Rice Husk to Improve Its Nutrative Value
by Rahat Naseer (2003-VA-196) | Dr. Abu Saeed Hashmi | Dr. Muhammad Tayyab | Prof. Dr. Habib ur Rehman.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Thesis submitted without CD. Availability: Items available for loan: UVAS Library [Call number: 2450-T] (1).
45.
Molecular Phylogeny And Diversity Analysis Of Bovidae (Boselaphus Tragocamelus, Antilope Cervicapra) And Cervidae (Axis Axis, Axis Porcinus) In Pakistan
by Ghulam Abbas (2011-VA-748) | Dr. Asif Nadeem | Prof. Dr. Mansoor Ellahi Babar | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Many species of mammals have declined within the past two centuries due to human
caused disturbances and the unsustainable use of natural resources. Molecular methods have an
important role in phylogeny and diversity analysis. The present study was designed for diversity
analysis of Boselaphus tragocamelus & Antelope cervicapra (Bovidae) and Axis axis & Axis
porcinus (Cervidae) family in Pakistan. A total of 25 samples from each of the four species were
collected from different parks, zoos and natural habitats. DNA was extracted, PCR primers were
designed and cytochrome-b, cytochrome-c gene and d-loop regions were amplified by PCR.
PCR products were sequenced bi-directionally by Big DyeTM Terminator. Bioinformatics tools,
Blast 2 sequences, Clustal-W, MEGA-6, Bioconductor in “R” were applied for analysis. The
clustering of the samples indicates that each species contains less within-population genetic
variability. Same pattern was observed when sequence of three genes was combined and MDS
plot was constructed. Phylogenetic analysis of the gene sequences revealed that each species
comprised a clade that is clearly distinct from the clade comprised of other species of deer
selected for this study. Finding of this study indicated that these species of deer have significant
genetic variations among-species that differentiate them from each other. This is the first report
from our region. The information of selected species of deer is prerequisite for designing
effective strategy in future conservation practices. However further genomic investigations
should be carried out at larger scale. Availability: Items available for loan: UVAS Library [Call number: 2560-T] (1).
46.
Identification Of Genetic Variants In The Low Density Lipoprotein Receptor Gene Causing Familial Hypercholesterolemia And Its Sequence Homology With Mus Musculus
by Ameer Hassan (2014-VA-504) | Dr. Wasim Shehzad | Dr. Muhammad Yasir Zahoor | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Familial hypercholesterolemia (FH) is caused by mutations in the genes coding for the low-density lipoprotein receptor (LDLR), apolipoprotein B-100, or proprotein convertase subtilisin/kexin type 9 (PCSK9). In this study, a molecular analysis of LDLR gene was performed in 20 unrelated patients from Pakistan. All patients were clinically diagnosed with definite or possible hypercholesterolemia according to a uniform protocol and internationally accepted WHO criteria. Preferable study was made to highlight the Genetic variation in Exon 4 of LDLR gene associated with defective catabolism of cholesterol effecting lipid metabolism which results in Familial Hypercholesterolemia. The extraction of genomic DNA was done from all selected blood samples. By selecting primers they were synthesized and optimized on extracted DNA samples. PCR product was sequenced and aligned. Mutations in the LDLR gene and its sequenced homology with Mus musculus were analyzed. We didn’t found any polymorphisms in the LDLR gene exon 4. So we concluded that there is no association between SNPs and increased levels of cholesterol in Pakistani population. More research should be carried out in Pakistan by increasing the sample size and considering the other regions of LDLR gene. This study will help the early detection and treatment of such cases and may ultimately reduce the incidence of mortality due to myocardial infarction. Apart from diagnosis, we also suggest it will be a potential therapeutic strategy to manage FH. Availability: Items available for loan: UVAS Library [Call number: 2538-T] (1).
47.
Polymorphism Analysis Within Tata-Box Of Bovine Lactoferrin Gene And Its Association With Mastitis In Sahiwal Cows
by Kashmala Haroon (2014-VA-04) | Dr. Sehrish Firyal | Dr. Immad Rashid | Dr. Muhammad Tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Mastitis is one of the most important diseases in dairy cows throughout the world, and is responsible for significant economic losses to the dairy industry especially in Pakistan. Several factors are responsible for this disease and about 20% bovines are suffering with this disease. Mastitis susceptibility and resistance is influenced by genetic variation of animals. Variations to polymorphisms in LF gene assume critical part of the immune response to mastitis. Polymorphism within LF gene may influence immune response to the mastitis in bovines. Recent study shows that promoter region of LF gene is highly polymorphic among bovines.
Present study was planned to identify polymorphism analysis within TATA-box of bovine LF gene and its association with mastitis. Multiple blood samples were collected from Sahiwal cows having clinical and sub-clinical mastitis. 10 samples were collected as a control. DNA extraction was done by organic extraction method and then quantification was done by Nanodrop. Amplification and sequencing was performed to get desire sequence of the gene. Comparative study of obtained sequence results were analyzed by using NCBI blast. Bioinformatics analysis was done with the help CLUSTAL W and BioEdit softwares.
Two novels and one reported SNPs were discovered within TATA-box of LF gene that might be having strong genetic association with mastitis in Sahiwal cows. This gene is strong candidate gene to differentiate between mastitis susceptible and resistant Sahiwal cows.
Availability: Items available for loan: UVAS Library [Call number: 2584-T] (1).
48.
Snp Genotyping Of Cacna1a Gene Implicated In Childhood Absence Epilepsy (Cae)
by Wajeeha Tariq (2010-VA-487) | Dr. Muhammad Wasim | Dr. Ali Raza Awan | Dr. Muhammad tayyab.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Childhood absence epilepsy (CAE) is more pediatric epileptic syndrome. It is about 5 to 15% of all childhood epilepsies. CAE is polygenic and multifactorial syndrome. Many different genes other than CACNA1A gene are involved to cause the CAE collectively. Mutation in P/Q type alpha 1 A subunit channel (Cav2.1) gene CACNA1A, leading to the reduction of Cav2.1 activity in both neurons and in expression system. Reduction in Cav2.1 channel activity altered the neurotransmitter release at neocortical synapses. Molecular genetics techniques have identified various mutation in the genes of ion channels such (CACNA1A, CACNA1G, CACNA1H, CACNB4), sodium channel genes (SCN1A, SCN1B and SCN2A) and GABA receptor genes (GABRD and GABRG2). CACNA1A ion channels are the standard mediator of neurotransmission in Central nervous system (CNS) and mutations in this gene play significant role in the generation of absence seizures. Pore forming alpha 1 a (Cav2.1) channels encoded by CACNA1A gene and are usually located in presynaptic neuron.
Present study was aimed to examine coding regions of CACNA1A gene for analyzing the mutations involve in epilepsy.
Blood samples (n = 40) of true CAE representatives were collected from Children hospital Lahore. DNA was isolated from all blood samples through standard organic method. Amplification of CACNA1A gene exon 36 regions was done with specially designed primers.
Later on, results were analyzed through sequencing of target region. Sequenced samples were analyzed through BioEdit software and alignment was done through Clustal Omega software.
It has been identified that absence epileptic patients of Pakistan showed Mutation in exon 36 of CACNA1A gene at position 281258bp and 281285bp which alter the protein sequence. Due to frame shift mutation a stop codon was detected at position 1813 in protein sequence. So a truncated and loss of function Cav2.1 channel might be formed. In epileptic patients, mutation is responsible for the absence seizures.
In the conclusion, we can say that additional study with large number sample is required to amend the effects of these mutations and their associated factors are precisely and perfectly identified. Further, there is need to investigate the other gene variation causing epilepsy in the local population of Punjab Pakistan. This study will ultimately help to develop genetic counseling strategies, gene therapies and parental diagnostic procedures for the Pakistani population.
Availability: Items available for loan: UVAS Library [Call number: 2746-T] (1).
49.
Production Of Polyhydroxybutyrate By Submerged Fermentation Using Agricultural By-Products
by Zainab Bibi (2015-VA-802) | Dr. Muhammad Tayyab | Dr. Shagufta Saeed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Increasing non-degradable waste on planet is the major environmental concern these days. Hence, there is an absolute need of “eco-friendly” plastics. Polyhydroxybutyrate (PHB) is the most popular biodegradable as well as eco-friendly polymer. However, high production cost of PHB is still a major problem in the commercialization of biodegradable plastics. Process economics revealed that the use of cheap and renewable carbon substrates such as agro-industrial wastes can account for 40-50% reduction in overall production cost.
In present study wheat bran, gram bran, rice bran, wheat straw and sesame oil cake were used to check PHB production using Azotobacter vinelandii NRRL-146641. For this purpose, 0.5ml inoculum was added in fermentation media and kept for incubation at 24-48 hrs. After incubation, both physical and chemical parameters such as (substrate water ratio, incubation time, inoculum volume, pH, agitation rate and nitrogen sources) were optimized. Optimized culture medium was centrifuged and obtained sediment was then used for analysis.
It was found that Azotobacter vinelandii in Rice bran contained medium gives maximum yield of PHB (248mg/100mL) at 8% substrate water ratio after 96 hours of incubation period (292mg/100mL), at 1.5 mL of volume of inoculum (304 mg/100mL), at pH 6.0 (316 mg/100mL), at 160 rpm agitation rate (416mg/100ml) at 0.3 % of yeast extract (446 mg/100mL) and 0.25% (436mg/100mL) of peptone. Obtained data was then analyzed by means of ONE-WAY ANOVA and through LSD test. Availability: Items available for loan: UVAS Library [Call number: 2855-T] (1).
50.
Evaluation Of Bioactive Peptides/ Proteins/ Alkaloids From Extracts Of Croton Tiglium, Lawsonia Inermis And Eruca Sativa Against Mastitis Causing Bacterial Strains
by Rubia Saeed (2011-VA-377) | Dr. Muhammad Tayyab | Dr. Abu Saeed Hashmi | Dr. Muhammad Nawaz.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Mastitis is considered as one of the most prevalent disease in dairy animals of Pakistan. Bacteria which are found in most mastitis cases are S. aureus, S. agalactiae and E. coli. Infections caused by these bacteria are being treated by various antibiotics but due to their development of resistance towards these drugs, there is need to explore some alternatives like medicinal plant extracts for the treatment of mastitis. Croton tiglium, Eruca sativa and Lawsonia inermis have been reported to have antimicrobial activity, thus the extracts of these medicinal plants will be explored to their antimicrobial activity against mastitis causing bacterial strains.
Present study purpose was to evaluate the bioactive proteins/alkaloids/peptides from extract of C. tiglium, E. sativa and L. inermis against mastitis causing bacterial strains. For this purpose, the leaves and seeds samples of selected medicinal plants (C. tiglium, E. sativa and L. inermis) were collected from Bagh-e-Jinnah and were identified from Department of Botany, University of the Punjab, Lahore. The ethanolic and aqueous extracts were prepared to evaluate the antimicrobial activity against mastitis causing bacterial strains. For this purpose, dust free leaves and seeds of selected plants were cut into small pieces, homogenized in ethanol/buffer and centrifuged. The resulting supernatant was then collected to check its antimicrobial activity against S. agalactiae, S. aureus and E.coli. Antimicrobial activity was analyzed by well diffusion method. Regarding the antimicrobial activity assay, the overnight grown cultures of the selected microbial strains was spread on the LB agar plates and the extracts was applied to wells incubated was done at 37°C for overnight. Inhibition zone was measured. Then the extracts having maximum activity were purified by GC.MS and the nature of extract was examined. All experiments were performed in triplicates so mean and average of the values was taken. Availability: Items available for loan: UVAS Library [Call number: 2853-T] (1).
