1.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Zingiber Officinale Rhizome Against Common Poultry Pathogens
by Ghalia Qayyum (2013-VA-779) | Dr. Aqeel Javeed | Dr. Qamar Niaz | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Antimicrobial compounds having plant origin inhibit bacteria through different mechanisms and can be used for the treatment of infections against resistant microbes. Majority of antibacterial drugs in clinical use are derived from natural origin. Hence, the present study is designed for antibacterial and cytotoxic evaluation of different extracts of Zingiber officinale rhizome against common poultry pathogens.
The four sequential i.e. hexane, chloroform, ethanol and aqueous extracts of Zingiber officinale were prepared by soxhlet extraction. Antibacterial activity of these extracts was determined by agar well diffusion method against Staphylococcus aureus, Clostridium perfringens type A, Escherichia coli, Salmonella enterica and Haemophilus paragallinarum. Zone of inhibitions were determined by well diffusion method. MICs of plant extracts were determined by micro broth dilution method. Cytotoxic activity was evaluated by applying MTT assay on Vero cell lines
The zone of inhibitions showed by hexane, chloroform, ethanol and aqueous extracts of Zingiber officinale against Staphylococcus. aureus were 12.33mm, 13.67mm, 16.33mm and 14mm; against Clostridium perfringens type A were 12mm, 16.33mm, 14mm and 8.33mm; against Escherichia coli were 14.33mm, 13.33mm, 14.33mm and 12mm; against Salmonella enterica were 17mm, 17.33mm and 12mm; against Haemophillus paragallinarum were 12.67mm, 13mm and 14mm respectively. Hexane extract showed no zone of inhibition against Salmonella enterica and aqueous extract was ineffective against Haemophillus paragallinarum.
MICs values of hexane, chloroform, ethanol and aqueous extracts of Zingiber officinale rhizome against Staphylococcus. aureus were 2500µg/ml, 625µg/ml, 2500µg/ml and 2083.33µg/ml; against Clostridium perfringens type A were 2500µg/ml, 312.5µg/ml, 1250µg/ml and 5000µg/ml; against Escherichia coli were 5000µg/ml, 1250µg/ml, 5000µg/ml and 5000µg/ml; against Salmonella enterica were 312.5µg/ml, 5000µg/ml, 5000µg/ml; against Haemophillus paragallinarum were 2500µg/ml, 1458.33µg/ml and 2500µg/ml respectively. MIC was not performed against hexane extract of Salmonella enterica and aqueous extract of Haemophillus paragallinarum as no zone of inhibition observed against them. Hexane extract of Zingiber officinale rhizome was cytotoxic at concentration ≥ 750µg/ml, chloroform extract at concentration ≥ 1500µg/ml and aqueous extract at concentration ≥5000µg/ml. Ethanol extract at concentration ranging from 1500µg/ml to 2.92µg/ml was not cytotoxic to cell.
The indigenous plant Zingiber officinale have antibacterial activity against common poultry pathogens and helpful to develop new drug from plant origin.
Availability: Items available for loan: UVAS Library [Call number: 2324-T] (2).
2.
Evaluation Of Comparative Antiviral Activity Of Indomethacin, Naproxen & Mefenamic Acid Against Avian Influenza H9 Virus
by Shahida Jamil Ahmed (2013-VA-850) | Dr. Aqeel Javeed | Dr. Muhammad Ovais Omer | Dr. Arfan Ahmad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Non-steroidal anti-inflammatory drugs play a vital role due to their multi therapeutic approach. In this study, the antiviral activity of indomethacin, naproxen, and mefenamic acid against avian influenza H9 virus was evaluated In ovo. The stock solutions of each drug were prepared in their perspective solvent and preserved. From the stock, three different dilutions (10µg/ml, 20µg/ml, 40µg/ml of indomethacin, 25µg/ml, 50µg/ml, 100µg/ml of naproxen and 20µg/ml, 40µg/ml, 80µg/ml of mefenamic acid) of each drug were prepared. For each of drug to be tested, 25 embryonated chicken eggs were assigned to 5 groups having 5 eggs each, to evaluate both antiviral activity and embryonic toxicity parameters.
For evaluating antiviral activity, the groups of embryonated chicken eggs were inoculated with 4HA virus, antibiotics and different concentrations of indomethacin, naproxen and mefenamic acid. For evaluation of embryonic toxicity, embryos of each group were injected with normal saline, antibiotics and different concentrations of indomethacin, naproxen and mefenamic acid. Two controls i.e. positive control of virus (received 4HA Virus only) and negative control (received normal saline) were also included to validate the test results.
With avian influenza H9 virus the different concentrations of each drug were mixed and 0.2 ml of this suspension was inoculated to 9th to 10th day embryonated eggs along with positive and negative controls having only virus and normal saline respectively. Amantadine, standard drug, was inoculated by following the mentioned manner. These inoculated embryonated chicken eggs were incubated at 37oC and were checked after 12 – 72 hours. After 72 hours of post inoculation, chilling was done by placing all the eggs at 4oC in fridge for overnight section of time and the allantoic fluid was collected.
The embryo survival percentage, positive or negative spot haemagglutination activity and determination of virus titre by haemagglutination test confirmed the antiviral activity. The embryonic toxicity effects of indomethacin, naproxen, mefenamic acid and amantadine were assessed by only inoculating the drug of respective concentrations as used for antiviral activity in embryonated chicken eggs and incubating for 72 hours. Among the three non-steroidal anti-inflammatory drugs (NSAIDs), indomethacin showed significant antiviral activity against influenza H9 virus as compared to naproxen and mefenamic acid. Naproxen showed antiviral activity against influenza H9 virus greater than that of mefenamic acid. However, antiviral activity of mefenamic acid as compared to naproxen and indomethacin is negligible against influenza H9 virus when confirmed by Spot Hemagglutination test while reduction in viral titre was observed by Hemagglutination test.
Availability: Items available for loan: UVAS Library [Call number: 2432-T] (1).
3.
Antibacterial And Cytotoxic Evaluation Of Sequential Extracts Of Astragalus Membranaceus Roots
by Sadia Alvi (2013-VA-595) | Dr. Aqeel Javeed | Dr. Muhammad Ovais Omer | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: The present study was designed to evaluate antibacterial and cytotoxic evaluation of different extracts of Astragalus membranaceus root against common poultry pathogens. Sequential extraction with hexane, ethanol, chloroform and aqueous solvents was prepared and antibacterial activity was evaluated by using agar well diffusion. Minimum inhibitory concentration (MIC) of plant extracts was evaluated by micro broth dilution test. The extracts exhibiting antimicrobial activity were further evaluated for cytotoxicity by using MTT assay on Vero cell line. Cell culture media was prepared and cell lines were propagated, monolayer was formed. This monolayer was exposed to plant extract dilutions. After 24-48 hours, MTT dye was introduced and cell survival percentage was calculated. Statistical analysis was conducted with Statistic Package for Social Sciences (SPSS for windows version 16, SPSS inc, Chicago, IL, USA). Results of antibacterial activity and MTT assay were compared using DMR posthoc test. Growth of Clostridium perfringens, Escherichia coli, Haemophilus species, Salmonella enterica and Staphylococcus aureus inhibited by all extracts of Astragalus except aqueous extract which shows no zones of inhibition against C. perfringes. MIC values were higher for aqueous extract against all selected bacteria and lowest for chloroform against E. coli, S. enterica and Staph. aureus (208.3ug/ml, 156.25ug/ml, 78.125ug/ml respectively) for hexane against Haemophilus species (833.3ug/ml) and for all three extracts against C.perfringes (1250ug/ml). Hexane, chloroform and ethanol extracts were appeared to be safe at all concentrations except ≥ 2000μg/ml, ≥1000μg/ml and ≥3000μg/ml respectively while aqueous extracts showed cytotoxicity at concentrations ≥625μg/ml. Astragalus membranaceus
SUMMARY
104
showed antibacterial activity against all selected pathogens. Chloroform and hexane extracts showed greater antibacterial activity than ethanol and aqueous. Cytotoxicity values for chloroform extract are safer than rest of three extracts. Astragalus membranaceus may be used to design traditional medicines for the development of therapeutic agent which will be more safe, effective and economical. Availability: Items available for loan: UVAS Library [Call number: 2444-T] (1).
4.
Effect Of Colchicine On Cellular And Humoral Immune Responses In Mice
by Shahzada Khurram Syed (2007-VA-444) | Dr. Aqeel Javeed | Prof. Dr. Muhammad Ashraf | Dr. Jawad Nazir | Dr. Shahbaz Yousaf.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Colchicine is a medication that treats gout. It is a natural product and secondary metabolite, originally extracted from plants Colchicum autumnale .It causes modulation of chemokine and prostanoid production and inhibition of neutrophil and endothelial cell adhesion molecules by which it interferes with the initiation and amplification of the joint inflammation.
The present study is designed to evaluate the effects of colchicine on cellular and humoral immunity in mice. There were five groups for each assay i.e. group I (negative control), positive control and three colchicine treated group II (40μg/kg), group III (80μg/kg) and group IV (160μg/kg). The number of mice in each group was five to eight. All these groups were administered doses intraperitoneally. To determine the effect of colchicine on cell mediated immunity , delayed type hypersensitivity (DTH) assay, macrophage engulfment assay, cyclophosphamide induced neutropenic test and nitric oxide production was performed .DTH was performed by measuring skin thickness. DTH showed significant difference (P<0.001) of negative control to colchicine treated groups 40μg/kg, 80μg/kg and 160μg/kg. With increasing dose, there was decrease in skin thickness of the mice. Highest reduction of skin was found at 160μg/kg. Macrophage engulfment assay was performed to evaluate the effect of macrophage induced phagocytosis. There was significant ( P <0.001) difference of engulfment of SRBCs by macrophages with negative control to colchicine treated group II (40μg/kg), group III(80μg/kg) and group IV(160μg/kg) groups. There was significant difference of engulfment of macrophages at 45 and 90 minutes.
Cyclophosphamide induced neutropenic test was performed to assess the effect of colchicine on total leukocyte count (TLC) and differential leukocyte count (DLC). There was
SUMMARY
77
reduction of TLC to about 45.3% in control to 48.3%, 54.68% and 65.42% in group II (40μg/kg), group III (80 μg/ kg) and group IV (160μg/kg) respectively when these were compared with primary values of TLC. There was significant difference of reduction in the neutrophil count of negative control 1057 (±120) to 902 (±67) in group II (40μg/kg), 734(±69) in group III (80 μg/ kg) and 609 (±71) in group IV (160μg/kg) of doses of colchicine. This test showed that with the increasing dose of colchicine, there was significant (P<0.001) difference of TLC count and neutrophil count.
Nitric oxide (NO) production by macrophages was performed for measuring different concentrations of nitric oxide produced. There was significant difference (P<0.001) in NO production by macrophages alone and LPS stimulated between negative control to group II (40 μg /kg), group III (80μg/kg), group IV (160μg/kg) of colchicine. With increasing dose, there was significant reduction in production of NO. There was significant P<0.0001 reduction in body weight andspleen weight difference of mice in different groups of colchicine treated 40μg/kg, 80μg/kg and 160μg/kg from negative control after treatment. There was difference of weight of Thymus of group II (40 μg/kg), group III (80μg/kg) and group IV (160μg/kg) but difference was statistically not significant. There were no histopathological changes observed in spleen and Thymus at 40μg/kg and 80μg/kg doses of colchicine. At 160μg/kg dose, increase in thickness of trabecular was seen .due to edema in the spleen. For evaluation of colchicine effect on humoral immunity, haemagglutination assay, mice lethality test and Jerne hemolytic plaque formation were performed. Haemagglutination assay (HA) was performed by using red blood cells injected intraperitoneally in mice to measure antibody titer. There was significant difference of (P >0.001) to colchicine treated group II (40μg/kg), group III (80μg/kg) and group IV (160μg/kg)with group I (negative control).With the increasing dose, there was reduction in the
SUMMARY
78
HA titer. Mice lethality test was performed by testing immune response of the mice to the challenge infection of P.multocida. It was performed by comparing mortality ratio of mice after administration of drug. There was no death of mice in the negative control group in which there was administration of PBS and vaccine. At 40μg/kg dose of colchicine, there was 50% mortality ratio. At 80μg/kg dose of colchicine 75% mortality ratio was observed. Maximum mortality ratio was observed at the 160μg/kg colchicine dose i.e. 100%.
Jerne plaque formation test was performed and plaques formed was enumerated and recorded as the number of plaque forming cells (PFCs) per million cells. There was significant difference (P<0.001) of reduction in number of plaques from negative control to all doses of colchicine 40 μg/kg, 80 μg/kg and 160μg/kg. Antibody formation was decreased with increasing the dose of colchicine. Therefore, it is concluded that colchicine suppresses the cellular and humoral responses in mice. Availability: Items available for loan: UVAS Library [Call number: 2650-T] (1).
5.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Zingiber Officinale Rhizome Against Common Poultry Pathogens
by Shumaila Nawaz (2013-VA-442) | Dr. Muhammad Adil Rasheed | Dr. Aqeel Javeed | Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Plants produce a diverse range of bioactive molecules, making them rich source of
different types of medicines. Calotropis procera, a giant milk weed, is known for its
pharmacological importance for centuries. This shrub has been known to possess analgesic,
antitumor,
antihelmintic,
antioxidant,
hepatoprotective,
anti-diarrhoeal,
anticonvulsant,
antimicrobial, oestrogenic, anti-nociceptive and anti-malarial activity. A very little information is
available regarding the antibacterial and cytotoxic activity of Calotropis procera so the present
study is designed to evaluate the antibacterial and cytotoxic activity of this plant. This study was
conducted to access antibacterial and cytotoxic activity of Calotropis procera.
Hexane, chloroform and ethanol, aqueous extracts were prepared by sequential extraction
method and antibacterial activity was evaluated against Staphylococcus aureus, Escherichia coli,
Salmonella enterica, Clostridium perfringens type A and Haemophilus paragallinarum by agar
well diffusion method in which inhibitory zones were measured. The extracts which showed the
antimicrobial activity were evaluated for cytotoxicity by using MTT assay on Vero cell line. Cell
culture media was prepared and cell lines were propagated, monolayer formed. Monolayer was
exposed to plant extract dilutions. After 24-48 hours, MTT dye was introduced and cell survival
percentage was calculated.
Statistical analysis was conducted with Statistic Package for Social Sciences (SPSS for
windows version 16, SPSS inc, Chicago, IL, USA). Results will be compared using one way
ANOVA analysis.
102
SUMMARY
Chloroform and ethanol extracts of Calotropis procera leaves have antibacterial activity.
It may help to design traditional medicines for the development of therapeutic agent which will
be more safe, effective and economical.
Availability: Items available for loan: UVAS Library [Call number: 2330-T] (1).
6.
Evaluation Of Antiviral Activity And Embryonic Toxicity Of Doxycycline, Ciprofloxacin Alone And In Combination With Ibuprofen Against Avian Influenza H9
by Aisha Nazir (2013-VA-851) | Dr. Muhammad Ovais Omer | Dr. Aqeel Javeed | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: This project was designed to analyze the antiviral and embryotoxicity of doxycycline,
ciprofloxacin alone and incombination with ibuprofen against H9 virus by using embryonated chicken eggs of 10 days old. The different concentrations of these agents were taken and two fold dilutions were made. Dilutions were mixed with avian influenza H9 virus and inoculated in embryonated eggs. Eggs viability was checked during incubation at 37°c temperature. After overnight chilling, haemagglutinition test was performed for evaluation of antiviral activity. Antiviral activity of these dilutions was calculated as embryo survival percentage and positive and negative hemagglutination activity. For embryotoxicity, dilutions were made in normal saline without virus and checked the results by mortality ratio after 48 hours of incubation.
The study provided information regarding antiviral activity and embryotoxicity of doxycycline, ciprofloxacin alone and incombination with ibuprofen at different concentrations. The present study showed that antiviral activity increased when used doxycycline and ibuprofen incombination. After using incombination it’s antiviral activity was high at these concentrations.
Results of antiviral analysis showed that doxycycline, ciprofloxacin and ibuprofen had mild antiviral activity alone and after using combination of doxycycline and ibuprofen the antiviral activity was increased. So these agents can be used as alternative therapy against avian influenza H9 virus. The outcomes were statistically analyzed by one-way ANOVA and Post-hoc Test was used to compare difference of means.
Comparative analysis of antiviral activity of doxycycline, ciprofloxacin and ibuprofen alone and in combination showed that doxycycline and ibuprofen when used incombination had
comparatively strong antiviral activity. It’s antiviral activity was stronger as compare when these agents used alone. In term of embryotoxicity these agents are not toxic. Availability: Items available for loan: UVAS Library [Call number: 2437-T] (1).
7.
Chemical, Microbiological And Toxicological Evaluation Of Textile Dyeing Industry Wastewater
by Muhammad Furqan Akhtar (2011-VA-265) | Prof. Dr. Muhammad Ashraf | Dr. Aqeel Javeed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Exposure to complex mixtures like textile effluent poses risks to animal and human health such as mutations, genotoxicity, pathological lesions and oxidative damage. The aim of the present study was to quantify metals and identify organic pollutants in untreated textile dyeing industry wastewater, to determine the bacterial load of wastewater, isolate and identify heavy metals tolerant bacteria and to determine its mutagenic, genotoxic and cytotoxic potential, influence on normal physiology and effects on oxidative stress biomarkers in effluent exposed rats.
Metal analysis through AAS revealed presence of high amounts of zinc, copper, chromium, iron, arsenic and mercury in industrial effluent. Various organic pollutants such as chlorpyrifos, cucurbitacin-b and phthalates were identified by screening through GC-MS.
Microbiological evaluation of textile dyeing industry wastewater revealed a high bacterial load. Different bacteria isolated from wastewater such as Staphylococcus aureus, Pseudomonas aeruginosa, Corynebacterium xerosis, Bacillus megaterium, Staphyoloccus epidermidis and Micrococcus varians exhibited resistance to Cr and Cu salts and antibiotics to varying degree.
Ames test with/without enzyme activation and MTT assay showed strong association of industrial effluent with mutagenicity and cytotoxicity respectively. Bacterial reverse mutation assay revealed that the mutagenicity of textile dyeing industry wastewater decreased with increase in dilution of wastewater.
In-vitro comet assay revealed the evidence of high oxidative DNA damage induced by textile wastewater. Wastewater exhibited concentration dependent genotoxicity in sheep
SUMMARY
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peripheral lymphocytes. When Wistar rats were exposed to industrial effluent in different dilutions for 60 days, then activities of total superoxide dismutase and catalase and hydrogen peroxide concentration were found to be significantly lower in kidney, liver and blood/ plasma of effluent exposed rats than control. Vitamin C at a dose of 50mg/Kg/day significantly reduced oxidative effects of effluent in rats. Industrial effluents may decrease activities of T-SOD and CAT and concentration of H2O2 in liver, kidney and blood/plasma of Wistar rats. Vitamin C may have a possible ameliorating effect on industrial effluent induced oxidative stress in Wistar rats.
Wastewater exposed rats exhibited necrosis of epithelial cells of nephron, pulmonary emphysema, and inflammation of the lungs, degradation and infiltration of cardiac myocytes, fibrosis of the liver, damage to the intestinal mucosa and sloughing off epithelial cells from the intestinal lumen.
This study concludes that untreated textile dyeing wastewater being a complex mixture of inorganic and organic pollutants may be highly eco-toxic and may contaminate of the environment via continuous release of various organic and inorganic pollutants. Availability: Items available for loan: UVAS Library [Call number: 2580-T] (1).
8.
Chemical Microbiological And Toxicological Evaluation Of Pharmaceutical Effluent Wastewater
by Ali Sharif (2011-VA-266) | Prof. Dr. Muhammad Ashraf | Dr. Aqeel Javeed | Prof. Dr. Aftab Ahmad Anjum .
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Pharmaceutical effluent being a complex mixture of drugs and heavy metals may affect human health exhibiting a strong potential of mutagenicity, carcinogenicity, cytotoxicity and oxidative stress induction along with pathological changes in various organs of the body. The current study was focused to quantify the presence of heavy metals, detection of various drugs, determining the bacterial load along with isolation and identification of different bacteria and assessment of the mutagenic and genotoxic, cytotoxic and oxidative stress induction of pharmaceutical effluent wastewater when exposed to sheep lymphocytes, Salmonella typhimurium strains, cell lines and rats respectively.
Atomic absorption spectrophotometer was used to quantify heavy metals and showed the presence of arsenic, chromium, lead and iron in concentrations above the normal limits recommended by WHO and EPA. Gas Chromatograph mass spectrophotometer analysis shown the presence of digitoxin, lignocaine, caffeine and trimethoprim and various other organic pollutants.
Microbiological evaluation showed a high bacterial load in the pharmaceutical waste water. Several bacteria were also found in PEW in the presence of different drugs and heavy metals. Aeromonas sobria, Micrococcus varians, Staphyoloccus epidermidis, Staphylococcus aureus, Bacillus megaterium showed tolerance to potassium di chromate and copper sulphate and resistance to various antibiotic discs.
Ames assay revealed a strong mutagenic potential with and without the presence of metabolic activation mixtures. A concentration dependent effect was observed when samples were tested with increasing dilution factor.
MTT assay and comet assay also showed a concentration dependent effect. The BHK-21 cell line was used to evaluate cytotoxicity and cell viability decreased with increasing concentration of PEW. Sheep lymphocytes used in comet assay exhibited a concentration dependent DNA damage.
Different antioxidant enzymes were also evaluated. Rats were exposed to PEW at different concentrations and following 60 days oral exposure, rats were evaluated for the presence of total superoxide dismutase, catalase and hydrogen peroxide in kidney, liver and plasma. Exposure to Pharmaceutical waste water significantly decreased the (TSOD), (CAT) and (H2O2) levels in plasma, liver and kidney. Treatment with Vitamin E significantly ameliorated the levels of enzymes.
Exposed rats were also evaluated for any pathological changes. Coagulative necrosis of renal epithelial cells were observed along with severe degeneration and cellular swelling in hepatocytes of hepatic cord.
Availability: Items available for loan: UVAS Library [Call number: 2600-T] (1).
