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Item Antimicrobial activity of nigella sativa plant leaves(UMT Lahore, 2018) Ayesha ChaudhryIn recent century, chemicals that used as anti-agents in medicines have become less active and more toxic. Due to this reason herbs have become vital source of clinical medicines because they have more resistant against organisms and are less toxic. Nigella sativa is a medicinal plant belong to family Ranunuclease. It contain many certain phytochemicals that exhibit activities i.e. antibacterial, antidiabetic, antifungal etc. This plant have been widely used to cure diseases. In this research, antimicrobial activity of N. sativa plant leaves Ethanolic extract was identified against bacteria (Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa & Aspergillus niger). Three antimicrobial techniques were applied: agar disk diffusion method, Agar well diffusion and Microdilution assay. Extract showed inhibition against gram negative bacteria but no inhibition seen in case of B. subtilis & A. niger. The extract did not inhibit the growth of gram positive bacteria. E. coli and P. aeruginosa has shown growth on media in presence of the extract. This study narrated that phytochemicals in leaves of Nigella sativa have potential to resist growth of gram negative bacteria. After further in vitro and in vivo trials, such extract can be used to treat ailments in animals and humans.Item Extraction purification and characterization of xylanase from bacillus subtilis(UMT Lahore, 2018) MASOOMA IQBALEnzymes play spearhead role in many industrial processes and lessen the cost that is why used for many years. Interest in xylanase has increased because of its applications and degrading abilities. It is an enzyme which is commonly found in all organisms such as bacteria, fungi and protozoans etc, whereas the biological system of mammals is unable to produce it. Xylanase hydrolyzes the molecules of the xylan into xylose monomers that are used by many bacteria and fungi as a source of energy. In the present study Bacillus subtilis producing xylanase enzyme were taken for production, purification and characterization. In this study xylanase was isolated from Bacillus subtilis strain and partial purification was done using Ammonium sulphate precipitation. DNS method was used to estimate the activity of the enzyme. Xylan was used as a substrate and xylose standard curve was constructed to check the activity of the xylanase. Cell free culture was saturated with 70% ammonium sulphate, centrifuged at 10,000 rpm and supernatant and precipitates were separated. Cell free sample, ammonium sulphate supernatant, ammonium sulphate precipitates and dialysis samples showed different activities which were 314.6, 245, 5.54 and 2.48 U/mg protein with enzyme activity 5.6, 3.6, 7.5 and 3.96 U/ml. Protein estimation of all the samples showed 178, 140, 21.6 and 14.52 mg/ml of proteins in respective samples. The enzyme was purified to 0.007 folds with 2.14 % recovery. Molecular weight was determined by SDS-PAGE which was 32kDa.Item Analysis of HBV and HCV prevalence in the general population of chaghi, balochistan(UMT Lahore, 2018) Jahangir AlamHepatitis viruses are threat for public health and considered as a depressing health issue globally. Hepatitis B virus (HBV) and hepatitis C virus (HCV) is main as well as leading reason for the liver cirrhosis and hepatocellular carcinoma development and they are also considered as one of the major reasons for TTIs (Transfusion Transmitted Infections) with considerable infection burden. Due to similar transmission mode the co-infection of both viruses are frequent. According to the reports of World Health Organization (WHO), there are 2 billion of HBV and 185 million HCV victims, which is 3% of the world’s population and three to four million adds to the list every year. The patients with co-infection of HBV and HCV have greater chances to develop further disease of liver. The medical assessment for the control of HBV and HCV viral diseases extremely needed because they both are affecting massive residents worldwide. The vaccine for HBV is approved when the techniques of suppression and elimination significantly improved. The current study consequences showed the prevalence of HBV and HCV in chaghi from 2001 to 2016. The samples are verified and analyzed by both test ICT and ELISA. Among the general population who visit the OPD, blood samples were collected from Sheikh Zaid Bin Sultan Hospital Chaghi, Balochistan, Pakistan. All the patients were the resident of District Chaghi and the studied time period of these patients are from June 2001 to July 2016. The prevalence of HBV in Chaghi within 17 years is 9.07% and prevalence of HCV is 6.9% out of 3288 individuals. There are many treatment options are available for the cure of HBV and HCV but still its number of victims per year is very higher and even deaths, because of hepatitis, are of great number. It can be prevented by taking simple precautionary steps, like never let anyone use already used syringe around you, avoid unsafe sex activities, keep the healthcare settings around you according to the international standards. The government should take serious steps for spreading the awareness of HBV/HCV virus and how they can be avoided. Along with Government, it is a moral responsibility of every citizen, who is aware of HBV/HCV virus, to play it’s role in preventing HBV/HCV around him/her and aware the people about it and the chain will keep growing and one day we will be able to beat this virus.Item Production, purification and characterization of asparaginase from bacteria E. Coli (Escherichia coli)(UMT Lahore, 2018) FATIMA IMANAsparaginase is an enzyme having (E.C. 3.5.1.1.), belongs to amidase group. This enzyme plays an important role in the treatment of tumor and non-Hodgkin’s lymphoma; it can act as anti-tumor agent, also necessary agent of chemotherapeutic agent against acute lymphoblastic leukemia. From last 50 years it has been used for the treatment of Acute Lymphoblastic Leukaemia (ALL), in the form of drugs. This was first identified from guinea pig serum in 1963. L-Asparaginase performs catalytic reaction, and involve in the conversion of asparagine to aspartic acid and ammonia. Both normal and tumor cells needs Asparagine for nutritional condition. Currently the clinical sources of ASNase are Erwinia chrysanthemi and Escherichia coli. The benefit in the use of this enzyme is that, it causes L-asparagine reduction; subsequently these amino acids are not synthesized by tumor cells. In a meanwhile there long term usage can causes allergic reactions, hypersensitivity and anaphylaxis. L-Asparaginase (L-AsnA) is broadly present in many microorganisms. This enzyme is also useful in different clinical research field like food industry, medicine and pharmacologic. The main objective of this research was isolation, production, purification and characterization of an extracellular L-Asparaginase from Escherichia coli. Production media were performed for asparaginase synthesis. By using the enzyme assay it has been characterized, having the principle of hydrolyzing l-asparagine into l-aspartic acid and ammonia. Enzyme assay proceed by nesslerization method. Crude enzyme of l-asparaginase was moderately purified by ammonium sulfate precipitation (70%) and dialysis. In the purification step, the specific activity of L-Asparaginase from E. coli was increases from 42.2 U/mg to 6.73 U/mg. In ammonium sulfate precipitation step, the total protein increased from 0.38 mg to14.54 mg. After that the last step is characterization where the SDS-PAGE was determined the molecular weight of the purified enzyme. Our thesis work exposed that only a single distinctive protein band of purified L-Asparaginase from E. coli was found the molecular weight is 32 kDa.Item An estimate of HBV and HCV seroprevalence in Muzaffarabad, AJK, Pakistan(UMT Lahore, 2018) Iffat RashidViral hepatitis is an issue affecting people all over the world. Around the globe hepatitis B virus (HBV) and hepatitis C virus (HCV) are the most important health issue. HBV and HCV are the most important cause of death and morbidity throughout the globe, despite the accessibility of vaccines and different treatment methods. These diseases (HBV and HCV) are major reason of chronic hepatitis, cirrhosis, and hepatocellular carcinoma worldwide. The co-infection of these viruses is frequent because they shared same modes of transmission. HBV and HCV co-infected patients have higher chances of movement, rapid fibrosis, and other liver disease. Patients that have dual infection of HBV and HCV, have higher chances of developing hepatocellular carcinoma (HCC), when compared to those who are mono-infected. The control of both of these viral diseases is tremendously important as they affect the enormous number of population globally. The procedures of eradication and elimination have improved considerably after the approval of vaccine against HBV and HCV. The testes were performed to verify the prevalence of HBV and HCV in Muzaffarabad Azad Jammu & Kashmir. Because of this reason male and female patients were registered in this study. The blood samples of 3,000 individuals who visited Sheikh Khalifa Bin Zyad and Combined Military Hospital, Muzaffarabad, Azad Kashmir from June 2016 to July 2017 were screened for HBV and HCV infection. Total 3,000 samples were collected and tested. Out of which 1,000 were male and 2,000 were female. ICT was used for antibodies findings. Among the 3,000 samples, 40 (1.33%) were shown HBV ICT positive result and 76 (2.53%) were shown HCV ICT positive result. To overcome these infections it is necessary to create awareness among the medical staff, subordinates and paramedics in the private and government areas. Physicians should be educated by giving short time trainings of active screening, initial treatment and outreach. Apart from awareness from the infectious disease there is a dire need of mass vaccination to reduce the current prevalence of HBV and HCV infection. The prevalence of HBV and HCV is expected to decrease by improved treatment strategy. Government should take different steps for giving reasonable treatment for these viral diseases, to avoid more infections. Early treatment to prevents chronic and liver cirrhosis and allows preventing liver transplantationItem Isolation and characterization of Begomovirus (ES) infecting ornamental plants(UMT Lahore, 2018) Syeda Kissa Ali GardeziBegomoviruses is a group of the viruses that affect plant production in different countries including Pakistan. Begomovirus is an emerging threat to dicot crops specifically in Punjab and Sindh. Cotton leaf curl disease (ClCuD) causes losses in billions to the economy of Pakistan thereby affecting 80-85% crop yield per year. Begomovirus genome consists of single stranded circular DNA (Deoxyribonucleic acid), transmitted exclusively by whitefly (Bemisia Tabaci). Identification of begomoviruses on weeds and fruit plants is important because they can serve as an alternative host and provide shelter, mixing vessel and inoculum source for next season field crops. Two Rhynchosia plant-leaf samples were collected from Johar town Lahore showing typical begomovirus symptoms. Total DNA was extracted using Cetyl Trimethyl Ammonium Bromide method. The DNA was subjected to polymerase chain reaction. No amplification was obtained using Beta01/02 and SON-A-F/R primer set. A 2.8 kb band size was obtained by using KTB-F/R primers. The sequence (identified as 26CG) was confirmed with sequencing. Sequence analysis showed that 26CG is a member of Rhynchosia yellow mosaic virus DNA-B (RYMV DNA-B). As predicted, Open Reading Frame (ORF) finder identified two orfs: NSP (Nuclear Shuttle Protein; 28.3 kDa) and MP (Movement Protein; 33.68 kDa). Sequence Demarcation Tool showed 96% identities of 26CG with RYMV DNA-B. Furthermore, Recombination Detection Program showed that 26CG may not be a recombinant due to its lower recombination score. Lastly, phylogenetic analysis performed with MEGA7 showed that it is a close relative of RYMV DNAB.Item Isolation and characterization of thermophilic bacteria(UMT Lahore, 2018) Ulfat Ali ShahIn assessing the bacterial diversity present in hot springs, extremotolerent and extremophiles bacteria (need tough environment for growth) were isolated. Several approaches were employed to identify bacteria and not survive in room temperature environment but exist in stressed environment. Samples were collected from Darkot and Hunza valley of Northern area of Pakistan. Temperature up to 4°C to 80°C and pH of neutral medium prior to selecting for extremophile bacteria. Hot springs are the best example of naturally existing extremophelic environment. In our research Bacillus Lichniformis_umt bacterial strain was isolated from the hot spring Hunza Murtazabad and Darkot where bacterial flora is not as much of explored. The thermophilic isolate was screened on the basis of various temperature of 37°C, 50°C and 60°C, and pH of 7 and 9 respectively. The biochemical characterization like sugar fermentation and chemical reactions studied compressively and phylogenetic tree was constructed and Bacillus Lichniformis_umt have 91% homology with neighbor specie strains. The isolate have potential applications in medicine, industry and biotechnology. This grows at optimum temperature of 50°C. It is sensitive to antibiotics of market include Leofloxacin, Erythromycin, HCL monohydrate, Azithromycin and Linofloxacin. This microbe can be a high good source to get industrially important thermostable enzymesItem Identification of anti-microbial activity of seed extract of nigella sativa(UMT Lahore, 2019-03-07) MARYAM ZAHRANigella sativa is known to be a medicinal remedy for several ailments hence its seeds’ extract and oil has been used in many experiments up till now. Decades have passed ever since the scientists have been working on it and experiments proved that there is almost no ailment against which it has not been proved effective. Nigella sativa seed oil is a cancer fighter, it protects brain damage due to lead poisoning, it is effective against liver problems, it protects kidney tissues, cures hypertension, prevents renal dysfunction and many more. Holy Prophet (PBUH) said that Nigella sativa (black seed) is an effective herbal remedy for every disease except death. In this experiment, seeds were used to prepare extract in ethanol and this extract was used as an antibiotic to identify antimicrobial activity against different strains of bacteria and fungi. Using different techniques and strains, like agar dilution method, wells in agar plates and disk diffusion method, experiments were done and results were evaluated. Certain quantities of antibiotic and strains were used. Strains could be bacterial as well as fungal but for the experimental techniques, would be different as same method would not show results for both types of strains. Results showed that the extract was effective against the bacterial strains and showed positive response in each technique that was used. However it was not entirely effective against the viral strains such as Aspergillus niger.Item Extraction and purification of lipase from aspergillus niger(UMT Lahore, 2018) NIMRA BABARLipase belongs to the serine class and catalyzes the hydrolysis of triglycerides to fatty acid and glycerol. Lipase is water soluble enzyme. Lipases have great importance in industry. Lipase used in different industries such as food processing, pharmaceutical, detergent, leather, cosmetics. Several plants, animals, fungi and bacteria are recognized to produce lipases. One of the major applications of enzymes is in durable detergents for household laundry. Lipase show different positional specificities and can catalyze different reactions like esterification, interesterification, and hydrolysis. Isolate the lipase from Aspergillusniger. Purification step was performed by ammonium sulfate precipitation and dialysis. Titrimetric method was used to estimate the specific activity of the enzyme. Bradford assay method was used to estimate the concentration of protein. SDS-PAGE was used to check the molecular weight of enzyme. The maximum extracellular lipase was purified 2.74 folds with an overall yield of 0.65% through purification procedure of ammonium sulfate precipitation. Maximum lipase activity of 0.23 U/ml and specific activity 21.9 U/ml was observed using olive oil as substrate. Protein estimation showed the concentration of enzyme present in the sample that was 420 mg/ml. Molecular weight determination was done by SDS-PAGE bands were not visualized on the gel, due to low concentration of protein,Item Isolation and characterization of culturable extremophile from potential areas of pakistan.(UMT Lahore, 2018) Aisha AshrafThe aim of this study was the isolation and characterization of extremophilic bacteria from the hot springs of Hunza from Gilgit-Baltistan, Pakistan. One microbe was isolated which showed optimum growth at 50°C at pH 7.0± 0.2. Antibiotic sensitivity of the microbes was tested against various antibiotics which included azithromycin, levofloxacin, lincomycin, ciprofloxacin and erythromycin. The microorganism showed sensitivity against all the above mentioned antibiotics. Furthermore, growth curve was checked using the UV spectrophotometer at absorbance of 600nm against time. The bacterial isolates were also tested for extracellular enzyme plate assay for amylase and protease, which exhibited positive and weak activity, respectively. Different biochemical tests including catalase, oxidase, nitrate reduction, gelatinase etc. were performed along with fermentation of different sugars and amino acids. When the biochemical characterization of the sample was performed, it revealed that it was Bacillus stearothermophilus. The study confirmed that the bacteria was a true thermophile which can provide a good source for biotechnological purposes. This microbe can be a potential source to get industrially important thermostable enzymes. The indigenous product of such enzymes can contribute in national development of the country.Item An estimation of hepatitis B virus prevalence in Skardu, Gilgit Baltistan(UMT Lahore, 2018) Ali RazaHepatitis B virus (HBV) is a main health issue all around the world. Regardless the availability of vaccines and various treatment strategies HBV is the main cause of mortality and morbidity throughout the World. The control of HBV is extremely important as it affects the huge number of individuals worldwide. The process of eradication and elimination of infection of HBV have improved significantly after the approval of vaccine against HBV. The pilot study was designed to check the seroprevalence of HBV in the capital of Baltistan Division. For this purpose male volunteer blood donors were enrolled in this study. In total 3377 individuals (aged between 20-30 years) who visited District Headquarter (DHQ) Hospital Skardu Gilgit Baltistan Pakistan from January to June 2016 were screened for HBV infection. The results of current study show the prevalence of HBV in Skardu from January to June 2016. In total 3377 individuals (aged between 20-30 years) who visited District Headquarter (DHQ) Hospital Skardu Gilgit Baltistan Pakistan from January to June 2016 were screened for HBV infection. The prevalence of HBV in the month of January was 1.7% out of 645 blood donors, February 3.36% out of 625, March 1.7% out of 648, April 4% out of 480, May 3.2% out of 220 and June was 5% out of 759. The overall prevalence of HBV in Skardu within six months is 3.16% out of 3377 blood donors. Awareness can be increased regarding HBV infection by educating the medical staff, medical subordinates and paramedics in the government and private sector. Primary physicians should be trained by giving short time trainings with respect to active screening, initial treatment and outreach. Apart from awareness from the infectious disease there is a dire need of mass vaccination to reduce the current prevalence of the disease. The prevalence of HBV is expected to decrease by improved treatment strategy. Early treatment of HBV prevents chronic HBV and liver cirrhosis and allows preventing liver transplantation. Government must take steps for providing reasonable treatment for HBV, which avoids more infections.Item Style guidelines for final year project reports isolation and characterization of extremophilic microorganisms from industrial effluents(UMT Lahore, 2018) Muneeba Waseem GhauriThe aim of this study was to isolate and identify extremophiles from the industrial effluent of Pakistan. Characterization of identified extremophile using biochemical and molecular approaches, their biotechnological potential in the production of extracellular enzymes and studying the phylogenetic affiliation of the extremophilic microbes in comparison with others. The range of temperature for the growth of the isolated strain ranged from 35°C to 37°C with its optimum growth at 37°C. Physiological analysis of strain were colonies in light yellowish cream color and spherical in shape. The range of pH for the isolated strain to grow ranged from pH 9 to pH 12 with its optimum growth at 37°C. Optimum NaCl and pH concentration for the strain to grow were 5% and 9.0, respectively. The full-length 16S rRNA sequence was 92.50% homologous to that of Alishewanella jeotgali strain MS1 and Alishewanella agri strain BAB6492.