1.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Opuntia Dillenii (Ker-Gawl) Haw. Leaves Against Common Poultry Pathogens
by Sadaf Raana | Dr. Muhammad Ovais Omer | Dr. Muhammad Adil Rasheed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: This project was designed to evaluate the antibacterial efficacy of hexane, chloroform, ethanol and aqueous extracts of Opuntia dillenii Haw. stems against common poultry pathogens. Pathogens used were Staphylococcus aureus, Escherichia coli, Salmonella, Clostridium perfringens type A and Haemophilus species.
This study was conducted to assess antibacterial and cytotoxic activity of O. dillenii Hexane, chloroform, ethanol and water extracts were prepared and antibacterial activity was evaluated by agar well diffusion method in which zones of inhibition were measured. Minimum inhibitory concentration (MIC) of plant extracts was evaluated by micro broth dilution method. 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 was formed. Monolayer was exposed to plant extract dilutions. After 24-48 hours, MTT dye was introduced and cell survival percentage was calculated.
O. dillenii stems extracts inhibited the growth of both Gram negative and Gram positive bacteria. Chloroform and ethanol extracts of O. dillenii showed significant antibacterial activity against all the pathogens studied as compared to hexane and aqueous extracts. Hexane extract showed maximum zone of inhibition against Haemophilus species (13mm), for chloroform extract maximum zone of inhibition was obtained for C. perfringens (25.6mm), for ethanol extract maximum zone of inhibition was obtained for C. perfringens (23.0mm) and for aqueous extract maximum zone of inhibition was obtained for C. perfringens (23.0mm). Minimum inhibitory concentration for chloroform extract was lowest for all the tested strains. For S. aureus, C. perfringens type A and S. enterica MIC was 1250μg/mL. For E. coli and
CHAPTER 6
SUMMARY
Summary
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Haemophilus species MIC was 2083.3 and 2916μg/mL, respectively. The extracts were further investigated to test cytotoxic effect on Vero cell line using MTT assay. Only ethanol extract was observed to be cytotoxic.
Statistical analysis was conducted with Statistic Package for Social Sciences (SPSS for windows version 16, SPSS inc, Chicago, IL, USA). The results of antibacterial activity and MTT assay were evaluated for significance of difference using analysis of variance (ANOV). The homogeneity of groups was verified by Duncan’s test at an alpha level equal to 5%.
Chloroform extract of O. dillenii stems possess antibacterial activity and can 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: 2281-T] (1).
2.
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).
3.
Antibacterial And Cytotoxic Evaluation Of Sequential Extracts Of Eucalyptus Globulus Leaves Against Common Poultry Pathogens
by Asma Iqbal (2013-VA-563) | Dr. Muhammad Adil Rasheed | Dr. Qamar Niaz | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Phytomedicines mark the major component of health care as natural medicines have always provided the strong foothold for the discovery and manufacturing of synthetic drugs. So plants are a rich source of bioactive compounds having many therapeutic activities and majority of them are still untapped. Eucalyptus globulus is a medicinal plant known for its value to cure asthma, respiratory infections, cough and allergic reactions. The antimicrobial activity, insecticidal and hypoglycemic activity have also been credited to the plant. Most of the studies have been conducted on the essential oils of Eucalyptus globulus and little work has been reported on extracts. Whereas, sequential extracts has not been employed yet.
Hexane, chloroform and ethanol, aqueous extracts were prepared by the sequential extraction on Soxhlet apparatus and antibacterial activity was evaluated against Staphylococcus aureus, Escherichia coli, Salmonella enterica, Clostridium perfringens type A and Haemophilus paragallinarum by agar well diffusion and micro broth dilution method. The zones of inhibition and minimum inhibitory concentration were determined. The extracts showing antimicrobial activity were further evaluated for cytotoxicity by using MTT assay on Vero cell line. The cell culture media was prepared and cell lines were propagated to form monolayer then monolayer was exposed to plant extract dilutions. After 24-48 hours, MTT dye was introduced and cell survival percentage was calculated.
The statistical analysis was conducted with help of Statistic Package for Social Sciences (SPSS for windows version 16, SPSS inc, Chicago, IL, USA) and results were compared using one way ANOVA.
Summary
89
The zones of inhibitions showed by hexane, chloroform, ethanol and aqueous leaf extracts of Eucalyptus globulus against Staphylococcus aureus were 0.0, 19.3, 20.3 and 23.3mm; against Clostridium perfringens type A were 14, 22.3, 14.0 and 15.3mm; against Escherichia coli were 0.0, 12.6, 13.3 and 15.6mm; against Salmonella enterica were 10, 12.3, 18.6 and 21mm; against Haemophilus paragallinarum were 0.0, 8.6, 14 and 18mm respectively. Hexane extract showed no zone of inhibition against Staphylococcus aureus, Escherichia coli and Haemophilus paragallinarum.
The MICs values of hexane, chloroform, ethanol and aqueous leaf extracts of Eucalyptus globulus against Staphylococcus aureus were 0.00, 104.1, 32.55 and 312.5 μg/ml; against Clostridium perfringens type A were 52.08, 39.06, 16.27 and 312.5 μg/ml; against Escherichia coli were 0.00, 78.12, 260.4 and 625.0 μg/ml; against Salmonella enterica were 13.02, 104.1, 130.2 and 416.6 μg/ml; against Haemophillus paragallinarum were 0.00, 104.1, 260.4 and 416.6 μg/ml respectively. MIC was not performed against Staphylococcus aureus, Escherichia coli and Haemophilus paragallinarum for hexane extract as no zone of inhibition was observed against them.
Hexane extract of Eucalyptus globulus was cytotoxic at concentration ≥ 312.5μg/ml, chloroform extract at concentration ≥ 375μg/ml, ethanol extract at concentration ≥ 625μg/ml and aqueous extract was cytotoxic at concentration ≥312.5 μg/ml.
The indigenous plant Eucalyptus globulus has antibacterial activity against common poultry pathogens and can be helpful for development of new drugs of plant origin. Availability: Items available for loan: UVAS Library [Call number: 2429-T] (1).
4.
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
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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).
5.
Antibacterial And Cytotoxic Evaluation Of Sequential Extracts Of Ocimum Basilicum Leaves Against Common Poultry Pathogens
by Shomaila Naz (2013-VA-1001) | Dr. Muhammad Ovais Omer | Dr. Muhammad Adil Rasheed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation 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 Ocimum basilicum seeds against common poultry pathogens.
The four sequential i.e. hexane, chloroform, ethanol and aqueous extracts of Ocimum basilicum leaves and seeds 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. All the results were statistically analyzed by one way ANOVA and compared means by Duncan’s multiple range of posthoc test at significance level of P≤0.05.
The results of zone of inhibitions showed by Ocimum basilicum leaves and seeds extracts ranging from 11.33-20.0 mm values of MIC results ranging from 4.889 μg/ml-2500 μg/ml of hexane, chloroform and ethanol. The aqueous extract of Ocimum basilicum have no activity against any bacterial pathogen. Ethanol extract of Ocimum basilicum leaves was cytotoxic at 500 μg/ml. Hexane extract of Ocimum basilicum seeds was cytotoxic at concentration ≥625 μg/ml, chloroform at concentration ≥19.53 μg/ml and ethanol extract at concentration ≥750 μg/ml.
The indigenous plant Ocimum basilicum 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: 2443-T] (1).
6.
Antibacterial And Cytotoxic Evaluation Of Different Extracts Of Glycyrrhiza Glabra (Liquorice) Roots Against Common Poultry Pathogens
by Javaria Arooj (2013-VA-596) | Dr. Muhammad Ovais Omer | Dr. Muhammad Adil Rasheed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: 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
Glycyrrhiza glabra Linn. roots against common poultry pathogens.
The four sequential i.e. hexane, chloroform, ethanol and aqueous extracts of Glycyrrhiza
glabra Linn. roots 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 and ethanolic extracts of
Glycyrrhiza glabra Linn. roots against Staphylococcus. aureus were10.3mm, 13.0mm, 11.6mm;
against Clostridium perfringens type A were20.0mm, 17.3mm, 17.3mm; against Escherichia coli
were11.6mm, 19.3mm, 16.0mm; against Salmonella enterica were13.6mm, 14.0mm,14.0mm;
against Haemophillus paragallinarum were13.0mm, 15.0mm, 17.0mm respectively. Aqueous
extract showed no zone of inhibition against any test bacteria.
MICs values of hexane, chloroform and ethanolic extracts of Glycyrrhiza glabra Linn.
roots against Staphylococcu aureus were 13.0μg/ml, 312.5μg/ml and 104.1μg/ml; against
Clostridium perfringens type A were 9.766μg/ml, 71.61μg/ml and 520.8μg/ml; against
Escherichia coli were 65.1μg/ml, 52.8μg/ml and 156.25μg/ml; against Salmonella enterica were
Summary
86
19.5μg/ml, 130.2μg/ml and 78.12μg/ml; against Haemophillus paragallinarum were 91.1μg/ml,
29.2μg/ml and 130.2μg/ml respectively. Aqueous extract showed no MIC value as no zone of
inhibitions wereobserved against them. Hexane extract of Glycyrrhiza glabra Linn. roots was
cytotoxic at concentration ≥ 650μg/ml, chloroform extract at concentration ≥ 2500μg/ml and
ethanolic extract was not cytotoxic to cell.
The indigenous plant Glycyrrhiza glabra Linn. roots 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: 2442-T] (1).
7.
Evaluation Of Antibacterial Effect Of Gymnema Sylvestre Species Cultivated In Pakistan
by Muhammad Tahir (2011-VA-339) | Dr. Muhammad Adil Rasheed | Dr. Qamar Niaz | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: This study was conducted to determine the use of medicinal plants as an antibacterial agent and its potential to as an alternative medicine against bacterial infections. For this purpose Sequential extracts (i.e. Hexane, Chloroform, Ethanol and Aqueous) of Gymnema sylvestre R.Br. were tested against S. aureus, E. coli, S. enterica, C. perfringens type-A, H. paragallinarum. Of each bacterium 3 isolates were tested by using well diffusion method. The results were obtained by determining the ZOI by well diffusion method and MIC by using 96 well ELISA plate.
The mean ZOI and mean MIC values of G. sylvestre leaves extracts showed that chloroform and ethanolic extracts have more antibacterial activity against all five microorganisms. Only chloroform and ethanolic extracts showed antibacterial activity against all 5 microorganisms while hexane extract showed antibacterial activity against S. enterica, S. aureus, H. paragallinarum, C. perfringens type- A but no activity was observed against E. coli. On the other hand aqueous extract have showed antibacterial activity only against C. perfringens type-A but no antibacterial activity against remaining four bacteria under study. While analyzing results based upon MIC, the chloroform extract has more antibacterial effect when compared with hexane. Hexane extract was more potent than aqueous extract whereas ethanolic extract was the least potent.
When overall antibacterial effect of all the extracts was evaluated against all bacterial strains, it was observed that C. perfringens type-A was the bacterium most vulnerable to antibacterial activity of sequential extracts of dried leaves of G. sylvestre as it responded
Summary
94
to all four sequential extracts and gave maximum zone of inhibition (10-22mm range) while no other bacteria showed such bigger zone of inhibition.
On the basis of MIC, it can be assumed that chloroform extracts have more antibacterial components as compared to hexane extract. Hexane extracts have more antibacterial components as compared to ethanolic extracts. The activity of aqueous extracts is negligible as it showed response against only one bacterium.
MTT assay was performed on supersaturated solutions of sequential extracts of dried leaves of G. sylvestre. Results revealed that small concentrations of these extracts are not toxic. Cell survival percentage (CSP) values below 50% were given at concentrations of 5800μg/ml (38.76%), 7225μg/ml (43.71%), 8150μg/ml (44.90%) and 3125μg/ml (41.84%) by hexane, chloroform, ethanolic and aqueous extracts respectively. Finally, on the basis of MIC and CSP for all of four sequential extracts, it is concluded that chloroformic extract is the most active and safe extract against all of 5 experimental bacteria, while hexane extract is safe against only C. perfringens type-A and ethanolic and aqueous extracts are cytotoxic on their MIC values for all the experimental bacteria. Statistical analysis showed that ZOI and MIC values were significantly different between the groups while within the same group they were non-significant.
Finally it can be concluded that the leaves of plant Gymnema sylvestre R.Br. cultivated in Pakistan has considerable antibacterial activity and considerable safety profile so it must be further studied, characterized, purified and chemically isolated so that may be converted to proper dosage form and this miracle plant may be used therapeutically to cure various ailments including bacterial infections especially poultry infections. Availability: Items available for loan: UVAS Library [Call number: 2503-T] (1).
