1.
Evaluation Of Antiviral And Cytotoxic Activity Of Medicinal Plants Extracts Against Infectious Bursal Disease Virus
by Waqas Ahmad | Prof.Dr.Muhammad Ashraf | Dr. Aqeel Javeed | Dr. Imran Altaf.
Material type: ; Format:
print
Publisher: 2011Dissertation note: The antiviral activity of plants Glyceriza glabra Linn. (roots), Phyllanthus emblicus Linn. (Fruit), Eugenia jambolana Lam. (Leaves), and Moringa oleifera Lam. (Leaves) were evaluated against Infectious bursal disease virus (IBDV) in this study. Ethanolic extraction of these plants was carried out by using Soxhlet apparatus and extracts was dried by using rotary evaporator. Four dilutions of each extracts viz 100, 50, 25 and 12.5?g/ml were made in distilled water. Vero cells were infected by mild strain of IBDV. Dilutions of these extracts were applied in triplicate manner on Vero cells that are confluent in 96 well cell culture plates. Positive control and negative control for antiviral assay were media plus cells and virus plus media respectively in antiviral assay. A cell culture plate was incubated for four days. After this incubation, viability of cells was determined by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] colorimetric assay. The cytotoxic activity of mentioned plant extracts was carried out by treating the cells with mentioned dilutions used in antiviral assay and incubating the 96 well cell culture plate for 4 days. Viability of cells was determined by MTT colorimetric assay. Positive and negative control for cytotoxic evaluation was cells plus media and cells plus media plus DMSO (10 %) respectively. Endpoint of this assay was measured in terms of cell survival percentage. Results were compared for qualitative variables using Chi-square technique and quantitative variables by linear regression analysis. 100 ug/ml and 50 ug/ml concentrations of Moringa oleifera Lam. showed cell survival percentages of 80% and 75% respectively and all four test dilutions of same plant showed no cytotoxicity for Vero cells. Two concentrations of Glycyrrhiza glabra Linn. 25ug/ml and 12.5ug/ml showed prominent cell survival of 75% and 80% respectively and other two concentrations 100ug/ml and 50ug/ml were found cytotoxic. Only 100ug/ml of Phyllanthus emblicus Linn. has shown cytotoxicity and 50ug/ml and 25ug/ml shown prominent antiviral activity. All concentrations of Eugenia jambolana Lam. were found non cytotoxic and 100ug/ml showed some antiviral potential against Infectious Bursal Disease virus.
Availability: Items available for loan: UVAS Library [Call number: 1319,T] (1).
2.
A Comparative Study Of Antiviral And Cytotoxic Activity Of Acacia Nilotica Against Peste Des Petits Ruminants
by Rizwana Raheel | Prof. Dr. Muhammad Ashraf | Dr. Aqeel Javeed | Dr. Imran Altaf.
Material type: ; Format:
print
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1449,T] (1).
3.
In Vitro Evaluation Of Antiviral And Cytotoxic Activity Of Ginseng Root, Leaves Of Tulsi And Aloe Vera Against Peste Des Petits Ruminants Virus
by Misbah Afzal | Prof. Dr. Muhammad Ashraf | Dr. Aqeel Javeed | Dr. Imran Altaf.
Material type: ; Format:
print
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1457,T] (1).
4.
Evaluation Of Mutagenicity And Cytotocicity Of Ferst Line Anti Tuberculosis Drugs
by Riffat Fatima | Dr. Muhammad Ashraf | Dr. Imran Altaf | Dr. Muhammad Adil Rasheed.
Material type: ; Format:
print
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1458,T] (1).
5.
Comparative Evaluation Of Mutagenicity And Cyhalothrin, Of Endosulfan, Lambda-Cyhalothrin,
by Umber Saleem | Prof. Dr. Muhammad Ashraf | Dr. Imran Altaf | Dr. Muhammad Ovais Omer.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1460,T] (1).
6.
In Vitro Antiviral Activity Of Leaves Extracts Of Azadirachta Indica, Moringa Oleifera And Morus Alba Against Foot
by Ishrat Younus | Prof. Dr. Muhammad Ashraf | Dr. Aqeel Javeed | Dr. Imran Altaf.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2012Dissertation note: The project was designed to assess in vitro antiviral and cytotoxic activity of leaves extracts of Azadirachta indica (AI), Moringa oleifera (MO) and Morus alba (MA) against Foot and Mouth disease virus (FMDV). Ethanolic, chloroformic and aqueous extracts of each plant were obtained by soxhlet apparatus. Chloroformic extracts were dissolved in cell culture media with the help of Dimethyl sulfoxide (DMSO). Eight concentrations 1 µg/ml, 6 µg/ml, 12 µg/ml, 25 µg/ml, 50 µg/ml, 100 µg/ml, 200 µg/ml and 400 µg/ml of each plant were used for both assays. Confluent BHK - 21 cells were grown in 96 well cell culture plates. Cells were treated by each concentration of extracts and extracts containing FMDV for cytotoxic and antiviral assay respectively in triplicate manner. Positive control (BHK-21 cells & cell culture media) and negative control (BHK-21 cells, FMDV & cell culture media) were kept for antiviral assay. For cytotoxic assay, positive and negative controls were kept as BHK-21 cells plus media and BHK-21 cells, media plus DMSO (20%) respectively. Cells viability and cytotoxic activity were determined by MTT assay for antiviral and cytotoxic assay respectively. Each extract was analyzed as cell survival percentage and expressed as means ± S.D. Statistical analysis was carried out by ANOVA. Seven plants extracts out of nine, exhibited antiviral activity against FMDV at a concentration non toxic to BHK-21 cell line. Ethanolic AI extract showed strongest anti-FMDV activity. Chloroformic MO leaves extracts showed significant antiviral activity. Chloroformic and aqueous MA leaves extract had no remarkable antiviral activity. At higher concentrations most of the plant extracts were cytotoxic
Availability: Items available for loan: UVAS Library [Call number: 1478,T] (1).
7.
Cytocenetic Effects Of Anti-Breast Cancer Drugs, Cyclophosphamide, Doxorubicin, Cisplatin And
by Zainab Batool | Prof. Dr. Muhammad Ashraf | Dr. Imran Altaf | Dr. Muhammad Ovais Omer.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2012Dissertation note: In this study mutagenicity and cytotoxicity of the chemotherapeutic agents used in breast cancer were evaluated. The drugs included in this study were Cyclophosphamide, Doxorubicin, Cisplatin and 5-Flourouracil. They were tested alone as well as in combination for their cytogenetic effects. The mutagenicity of these drugs was tested by Ames test using two strains of Salmonella i.e. TA100 and TA98 with and without S-9 at different concentrations. While for cytotoxicity evaluation MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) colorimetric assay was selected. 96 well plate and BHK-21 cell lines were used to perform this assay.
This study indicated that cyclophosphamide was mutagenic ( 62.5 µg/plate) to TA 100 with S-9 but non mutagenic to TA 98 with and without S-9, while the concentration of 250µg/ml and above was found cytotoxic. Doxorubicin was mutagenic to TA 100 and TA 98 with and without S-9 at 1 µg/plate and above, while cytotoxic dose was 10µg/ml and above. 5-FU was found non mutagenic in this assay to both test strains with and without S-9 at all test concentrations, however it was found cytotoxic above 5µg/ml in MTT assay. Cisplatin showed mutagenicity to both test strains at 2µg/plate and above , while at 5µg/ml and above it was found cytotoxic.
When the combinations of these drugs were tested for cytogentic effects , it was found that the concentrations which were non mutagenic individually became mutagenic and cytotoxic when combined together.
Availability: Items available for loan: UVAS Library [Call number: 1481,T] (1).
8.
Cytotoxic And Antiviral Evaluation Of Different Opuntia Species Against Peste Des Petits Ruminants Virus In Vitro Cell
by Faryal Ashraf | Prof. Dr. Muhammad Ashraf | Dr. Aqeel Javeed | Dr. Imran Altaf.
Material type: ; Format:
print
Publisher: 2012Dissertation note: The antiviral activities of Opuntia delinii, Opuntia manocantha, and Opuntia stricta were evaluated against Peste des petits ruminants virus (PPRV) in this study, as these plants are associated with a lot of antiviral activity as shown by literature review. Ethanolic and aqueous extracts of all the three species of Opuntia were obtained by using soxhlet apparatus (Davey et al. 2010) but first crushed them into small pieces with a sharp knife to have better extraction results. The resultant extracts were dried in rotary evaporator using standard operating procedures until semisolid extract was obtained. Different dilutions were made by dissolving in double distilled water. Vero cells were made mildly affected by mild strains of Peste des petits ruminants virus. Dilutions of these extracts were applied on Vero cell line in triplicate manner that was first made confluent up to 90% in 96 well cell culture plates. For performing anti viral assay, Positive control and negative controls used were media plus cells and virus plus media respectively. These plates were incubated for a period of four days. After this incubation period, viability of cells was determined by MTT colorimetric assay i.e. number of living and dead cells. The cytotoxic activity of above mentioned three plant species was performed by treating the Vero cells with different dilutions as used in antiviral assay and incubating the 96 well plates for 4 days. Viability of cells was determined by MTT assay. The positive and negative control for cytotoxic evaluation was cells plus media and cells plus media plus DMSO (Dimethyl sulfoxide) 5% respectively. Results were calculated in terms of cell survival percentage (CSP) for anti viral
and death rate (%) for cytotoxic assay. At highest concentrations, i.e.500 to 1000 µg/ml, all the ethanolic and aqueous extracts obtained from all the plant species showed cytotoxicity but at the lower concentrations ranging from 7.81µg/ml to 125µg/ml, there was no cytotoxicity. Antiviral and cytotoxic activity of the plant extracts was evaluated by applying Analysis Of Variance (ANOVA) and comparison between two extracts was performed by applying T-Test for statistical analysis. Statistically when these results were interpreted, they were insignificant because P value is more than 0.05. This research project has a lot of positive outcomes and future prospects. The extract of plants having good antiviral activity and with no cytotoxic activity will be good baseline for further evaluation.
CHAPTER 6
SUMMARY
The antiviral activities of Opuntia delinii, Opuntia manocantha, and Opuntia stricta were evaluated against Peste des petits ruminants virus (PPRV) in this study, as these plants are associated with a lot of antiviral activity as shown by literature review. Ethanolic and aqueous extracts of all the three species of Opuntia were obtained by using soxhlet apparatus (Davey et al. 2010) but first crushed them into small pieces with a sharp knife to have better extraction results. The resultant extracts were dried in rotary evaporator using standard operating procedures until semisolid extract was obtained. Different dilutions were made by dissolving in double distilled water. Vero cells were made mildly affected by mild strains of Peste des petits ruminants virus. Dilutions of these extracts were applied on Vero cell line in triplicate manner that was first made confluent up to 90% in 96 well cell culture plates. For performing anti viral assay, Positive control and negative controls used were media plus cells and virus plus media respectively. These plates were incubated for a period of four days. After this incubation period, viability of cells was determined by MTT colorimetric assay i.e. number of living and dead cells. The cytotoxic activity of above mentioned three plant species was performed by treating the Vero cells with different dilutions as used in antiviral assay and incubating the 96 well plates for 4 days. Viability of cells was determined by MTT assay. The positive and negative control for cytotoxic evaluation was cells plus media and cells plus media plus DMSO (Dimethyl sulfoxide) 5% respectively. Results were calculated in terms of cell survival percentage (CSP) for anti viral
and death rate (%) for cytotoxic assay. At highest concentrations, i.e.500 to 1000 µg/ml, all the ethanolic and aqueous extracts obtained from all the plant species showed cytotoxicity but at the lower concentrations ranging from 7.81µg/ml to 125µg/ml, there was no cytotoxicity. Antiviral and cytotoxic activity of the plant extracts was evaluated by applying Analysis Of Variance (ANOVA) and comparison between two extracts was performed by applying T-Test for statistical analysis. Statistically when these results were interpreted, they were insignificant because P value is more than 0.05. This research project has a lot of positive outcomes and future prospects. The extract of plants having good antiviral activity and with no cytotoxic activity will be good baseline for further evaluation.
Availability: Items available for loan: UVAS Library [Call number: 1493,T] (1).
9.
Genotoxicity And Mutagenicity Of Metformin And Aspartame Alone And In Combination
by Amna Nazar | Prof. Dr. Muhammad Ashraf | Dr. Aqeel Javeed | Dr. Imran Altaf.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1690,T] (1).
10.
Cytotoxic, Mutagenic And Genotoxic Evaluation Of Different Aesthetic Colorants
by Wardah Naeem | Prof. Dr. Muhammad Ashraf | Dr. Imran Altaf | Dr. Muhammad Adil Resheed.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2013Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1743,T] (1).
11.
Genotoxic and Mutagenic Potential of Anti-Diabetic Drugs (Sitagliptin and Metformin) Alone And In Combination With Artificial Sweeteners.
by Komal Najam | Prof. Dr. Muhammad Ashraf | Dr. Imran Altaf | Dr. Muhammad Adil Rasheed.
Material type: ; Format:
print
Publisher: 2013Dissertation note: Metformin most commonly prescribed oral anti hyperglycemic drug for type 2 diabetes whereas Sitagliptin recently approved oral antidabetic drug for type 2 diabetes were evaluated for their mutagenic and genotoxic potential alone and in combination with three artificial sweeteners (Saccharin, Aspartame and Acesulfame-K) normally consumed by diabetic individuals.
In this research project Ames Salmonella/microsome assay was performed to check the mutagenicity of Metformin and Sitagliptin alone and in combination with artificial sweeteners using mutant Salmonella tester strains TA100 and TA98 with and without the S9 whereas Genotoxicity was evaluated by Single Cell Gel Alkaline Electrophoresis/Comet.
The results indicated that Metformin alone showed mutagenic effect at 120µg/plate against TA100 with S9mix. However Metformin when tested in combination with artificial sweeteners, significant enhance in mutagenicity occurred only against TA100 with and without S9. Sitagliptin displayed mutagenic potential only to TA98 with S9mix at the concentration of 3040µg/plate. In addition significant enhance in mutagenicity occurred when tested in combination with artificial sweeteners against both strains with and without S9. In case of genotoxicity both Metformin and Sitagliptin results indicated significant increase in DNA damage in dose dependant manner as compared to negative control. Though Metformin and Sitagliptin in combination with artificial sweetener did not reveal any significant boost in the genotoxicity relative to when they were tested alone.
Availability: Items available for loan: UVAS Library [Call number: 1799,T] (1).
12.
Characterization And Use Of Phytase Producing Bacterial Isolates To Enhance The Nuteitive Value Of Poultry Feed
by Sohail Aslam | Prof, Dr. Ahmad anjum | Dr. Imran Altaf | Dr. Muhammad Wasim.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1930,T] (1).
13.
Genotoxic And Mutogenic Study Of Formaldehyde, Sodium Hypochlorite And Cresol
by Ann Fatima (2012-VA-995) | Prof.Dr. Muhammad Ashraf | Dr. Muhammad Adil Rasheed | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Disinfectants are used to control, prevent or destroy harmful microorganisms on inanimate surfaces. These chemicals are being used to dispose the contagious hospital wastes like disposable plastics and microbiological waste. Various factors like temperature, contact period concentration of disinfectant, organic soil and nature of water used for dilution affect of disinfection process. So, disinfectants must be tested prior to any specific applications for its proper effectiveness. This study has been designed to study the genotoxicity and mutagenicity of three commonly used disinfectants, formaldehyde, sodium hypochlorite and cresol alone and in combination.
Different dilutions of formaldehyde (1, 0.3, 0.1, 0.006, and 0.003%) sodium hypochlorite (8, 4, 2, 1, and 0.5%) and cresol (7.6, 3.8, 1.9, 0.95, and 0.475%) alone and in combination were investigated for mutagenicity as well as genotoxicity in vitro. Mutagenicity was investigated by Ames Salmonella/Microsome assay with and without metabolic activation system; S-9 with the help of two strains of Salmonella typhimurium, TA 100 and TA 98 and genotoxicity was checked by Comet assay using peripheral blood lymphocytes. The results were analyzed by statistical package of Social Sciences; results were presented as mean ± S.D and the data analysis was done by using one-way analysis of variance. Differences were considered significant at P < 0.05.
Summary
83
Formaldehyde, sodium hypochlorite & cresol showed significantly mutagenic potential against TA 100 & TA 98 strains of Salmonella with and without metabolic activation system and genotoxic effects. A higher concentration showed more significant results.
Formaldehyde, sodium hypochlorite & cresol has both mutagenic and genotoxic potential. But this mutagenicity and genotoxicity has been observed more with higher concentrations as compared to low concentration. Availability: Items available for loan: UVAS Library [Call number: 2305-T] (1).
14.
Comparitive Immunogenic Evaluation Of Purified And Whole Culture Vaccine Of Pasteurella Multocida B2 Of Bovine Origin
by Anika Khalid (2006-VA-357) | Dr. Imran Altaf | Dr. Jawad Nazir | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Pasteurella multocida is responsible for causing Heamorrhagic Septiceimia which is a fatal and acute disease mainly associated with bovines.
Pasteurella multocida consists of two important antigenic structures i-e Outer membrane protein(OMP) and Lipopolysaccharide(LPS) that forms capsule.These two structures played vital role in pathogenicity as well as in the immunogenicity.
In our present project we study the role of LPS,OMP alone and in combination as an immunogens.We compared the immunogenic behaviour of whole culture vaccine(consists of LPS and OMP) purified vaccine(consists of OMP),LPS vaccine (consists of LPS only) by IHA and CFT (using OMP and LPS both as an Ag separately) and the overall results showed that whole culture HS Alum precipitated vaccine produced better antiboby titer against both outer membrane proteins (OMP) and lipopolysacharrides (LPS) when checked against respective antigens as compared to the vaccines containing OMP and LPS alone.
So it was concluded that LPS no doubt act as an immunogen but its immunogenicity increases many times when combine with protein (OMP) as in whole culture vaccine. Availability: Items available for loan: UVAS Library [Call number: 2316-T] (1).
15.
Genetic Diversity Among Different Isolates Of Pasteurella Multocida From Poultry
by Arslan Sardar (2013-VA-282) | Dr. Imran Altaf | Prof. Dr. Aftab Ahmad Anjum | Dr. Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Fowl cholera is an acute bacterial disease of broiler breeders and layer breeders caused by Pasteurella multocida. In the present study, 10 isolates from different areas of Punjab were purified. These samples were confirmed by API Kit. Different molecular techniques like PCR and RFLP were used to investigate variation at the molecular level among 10 isolates collected from different areas of Punjab. Different mutations were observed among 10 field isolates at different mutation sites by sequencing. Phylogentic tree was also made using MEGA6 software that supported the sequencing results. ‘Msp1’ endonuclease cleaved bacterial whole genome at different cutting sites, all 10 isolates collected from different districts of Punjab cleaved into 3 to 5 fragments ranging from 600 to 10000 base pairs which showed the genetic variation among 10 isolates of P.mulocida. Availability: Items available for loan: UVAS Library [Call number: 2315-T] (1).
16.
Assessment Of Genotoxicity Of Propofol, Thiopental And Ketamine In Patients Under Balanced Anesthesia With Isoflurane
by Maidah Mehtab (2013-VA-597) | Dr. Muhammad Adil Rasheed | Dr. Tanveer Akhter Butt | Dr. Muhammad Ovais Omer | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Exposure of anesthetic agents to the patients and operating room staff may involve a genotoxic risk so the knowledge of their effects on genetic material can give valuable support to anesthesia care providers to make better treatment performance and improve patient safety.
Comet assay was used to study the genotoxic actions of three IV anesthetic agents (propofol, thiopental and ketamine) that were used for induction during balanced anesthesia with inhalational anesthetic isoflurane. Three groups consisted of total18 patients who were undergone elective abdominal procedure lasted about 2 hours. Intravenous samples of blood were obtained before anesthesia induction (T0 —baseline), immediately after anesthesia induction (T1), 10 min (T2), 60 min (T3) 120 min (T4), 6 hours (T5) and 12 hours (T6) after anesthesia induction. Lymphocytes were isolated and single-cell gel electrophoresis/comet assay was used in which the cell suspension on agarosed slides was lysed in high salts and detergents containing lysing solution, exposed to alkaline buffer solution for DNA unwinding and then following electrophoresis at 24 volts and 300 mA and stained with ethidium bromide. These preapared slides were analyzed under fluorescent microscope. The anesthetics induced damage to DNA on 50 cells per sample per patient was measured as total comet length (i.e. damage index) categorized as undamaged to highly damaged (class0- class3) cells.
The data collected was analyzed by analysis of variance (ANOVA) Post Hoc Test LSD using Statistical Package of Social Sciences (SPSS).
By comparing the genotoxicity of propofol, thiopental and ketamine, it can be concluded that propofol causes the least or no genotoxicity during balanced anesthesia with isoflurane and could be the best choice for induction when isoflurane is used for maintenance.
Availability: Items available for loan: UVAS Library [Call number: 2323-T] (1).
17.
Analysis Of Genetic Polymorphism In Exon 6 & 11 Of Glucosidase Beta Acid (Gba) Gene In Gaucher Diseased Patients From Punjab, Pakistan
by Aysha Arshad (2009-VA-571) | Dr. Muhammad Yasir Zahoor | Dr. Muhammad Imran | Dr. Imran Altaf.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Gaucher disease (GD) is amajor predominant heterogenic, inherited and metabolic lysosomal storage disorder. It is prompted by an alteration in glucosidase acid beta (GBA) gene. GBA gene encodes a 497 amino acid glucocerebrosidase enzyme. It is a lysosomal hydrolase, present in all mammalian cells membrane that carries the catalysis of complex ubiquitous sphingolipids called glucocerebrosides (GlcCer) into smaller and simpler molecules of sugar and ceramide. The human glucocereborside (GBA) gene is present in highly gene dense area on q arm of 21 chromosome and its fragment length is 7.8kb comprising of 11 exons. A pseudogene is also present in vicinity of GBA gene which shares 96% homology of sequence with functional gene. Genetic recombination and gene conversion among these two GBA genes are responsible for 10-20% GD mutations. >300 mutations of GBA have been described till 2014. GD has three different clinical forms depend on its heterogeneity. These are characterized by the age of onset and with or without the participation of CNS.
In this study, 10 blood samples were collected of GD patients from repository at Molecular and Genomic Laboratory located at IBBT department, UVAS Lahore and from Children Hospital Lahore. DNA extraction was done by using organic method from blood samples. Amplification of GBA gene exons 1, 6 and 11 was performed using PCR. PCR products were sequenced using Sanger di-deoxy sequencing method. Different bioinformatics tools were applied for the sequence analysis of exon 1, 6 and 11. We found two variants of GBA gene. A deletion of CT nucleotide repeat in intron 1 was found. We also found a substitutional change of nucleotide T>A in intron 8. Availability: Items available for loan: UVAS Library [Call number: 2334-T] (1).
18.
Assessment Of Microbial Load, Protease Activity And Aflatoxin M1 In Raw And Uht Milk Procured From Local Markets Of Lahore
by Sadaf Almas (2007-VA-250) | Dr. Imran Altaf | Dr. Ali Ahmad Sheikh | Dr. Muhammad Nasir.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Pakistan is among the largest milk producing countries. The requirement of milk is
increasing day by day. It has rapidly increasing demand and competition in national and
international markets. Milk consumers in Pakistan often face low-quality, lack of hygiene and
absence of cold chains as primary contributors to this low quality. Milk and dairy products also
become contaminated during manufacturing and packaging processes. The consumption of low
quality milk may cause milk borne diseases. Not only the bacteria but the presence of their
enzymes also can cause deterioration of the milk quality. The heat stable enzymes can cause the
spoilage of commercial UHT products without presence of any viable count. Aflatoxin (M1) is
metabolite of AFB1 in milk. It causes chronic diseases and immunosuppression in children. It is
found carcinogenic and cytotoxic in nature.
In this project microbial load, free amino acid estimation to predict any protease activity
and Aflatoxin M1 were studied in both UHT and Raw milk samples (n=15) procured from local
markets of Lahore. Three UHT brands A, B and C were purchased. The UHT milk was studied
for microbial growth and protease activity at purchase and at expiry of the products. The
microbial load was evaluated by testing of milk for Total viable count, Coliforms, Yeast and
Molds, Anaerobic Clostridia and Bacillus cereus in both raw and UHT milk. Protease activity
was estimated by assessing the free amino acid by using ninhydrin assay while the Aflatoxin M1
was detected through High performance liquid chromatography.
SUMMARY
80
CONCLUSION:
It was found that the locally available raw milk quality was poor for consumption and dairy
processing for safe and stable milk products. UHT milk quality was found better with low
microbial load. Protease activity with reference to free amino acid was detected in raw milk
which is indication of the poor milk storage conditions, cold chain maintenance and
unavailability of fresh milk. Protease activity was also found in UHT milk and an increase in free
amino acid which could be due to heat stable proteases active during shelf life of the milk
brands. Aflatoxin M1 was found in majority of raw milk sample which showed the poor animal
feed storage and monitoring system. Aflatoxin M1 was also found in some samples of UHT
brands with high concentration which depicted that AFM1 was heat stable and it retained in the
commercial UHT products as well. Availability: Items available for loan: UVAS Library [Call number: 2354-T] (1).
19.
Comparison Of Two Imported Live Attenuated PPR Vaccines In Local Sheep In Pakistan
by Saliha Saba | Prof. Dr. Aneela Zameer Durrani | Dr. Hassan Salem | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Peste des petits ruminants (PPR) also famous as goat plaque is of viral origin and is extremely contagious disease of sheep and goat (Dhar et al. 2002; Asim et al. 2009). PPR can cause high mortality about 50 – 80 % in non-immunized sheep and goat population. Due to its similarity with other diseases, Peste des petits ruminants (PPR) is being devalued but at the same time it is said to be one of the major constraints to successful small ruminant farming in tropics (Sen et al. 2010). PPR virus is paramyxovirus, enveloped and belongs to the genus morbillivirus. These viruses comprise of 16Kb long, single stranded RNA showing negative polarity (Barrett et al. 2005).
The various vaccines like homologous and recombinant vaccines have been manufactured for the management of Peste des petits ruminants (PPR), as no accurate treatment is available for its control. For the immunity of animals against this disease, the tissue culture based, attenuated rinderpest vaccine (TCRV) had been accustomed over a extensive period because of the antigenic association among RPV and PPRV (Diallo et al. 1989).With the help of fresh freeze-drying methods and stabilizing agents the thermostability of the present PPR homologous vaccine has been enhanced significantly (Worrwall et al. 2001).
In Pakistan, PPR vaccine was manufactured with the help of PPRV Nigerian 75/I (PPR 75/1 LK 6 Vero 75) for the sheep and goat immunization (Asim et al. 2009). India had manufactured numerous live attenuated vaccines like the PPRV Sungri/96 that has been regularized for use (Hegde et al. 2008). ). The Peste des Petits Ruminants (PPRV-Sungri/96 ) vaccine is being manufactured on small and large scale for prevention of Peste des Petits Ruminants (PPR) outbreaks in India (Singh et al. 2004).
Summary
41
The current study was designed to study the immunogenicity of two imported live attenuated PPR vaccines in local sheep. A total of sixty (60) animals were selected and further separated into two groups, viz. Group-A and Group-B, having thirty (30) animals each. Group-A was further sub-divided into A1 comprising 10 sheep to which Raksha PPR vaccine (Sungri 96) was administered, A2 comprising of 10 sheep to which PPR vaccine (Nigeria 75/1) was administered and A3 comprising of 10 non-vaccinated sheep which served as control. Group B was separated into two sub-groups i.e B1 and B2 having fifteen (15) animals each. The Group-B1 was sub-divided into B1a having 05 sheep to which Raksha PPR vaccine (Sungri 96) was only administered, B1b having 05 sheep to which along with Raksha PPR vaccine (Sungri 96), Vitamin AD3E was administered and B1c having 05 unvaccinated sheep which served as control.
Similarly the Group-B2 was sub-divided into B2a having 05 sheep to which PPR vaccine (Nigeria 75/1) was only administered, B2b having 05 sheep to which along with PPR vaccine (Nigeria 75/1), Vitamin AD3E was administered and B2c having 05 non-vaccinated sheep and served as control group respectively. The serum samples were collected and mean antibody titer was calculated by complement fixation test (CFT) at zero day, 7th day, 14th day, 28th day and 48th day post-vaccination.
The live attenuated, Raksha PPR (Sungri 96) vaccine induced the mean antibody titers of 0 ±0.00, 4.7±0.48, 4.7±0.48, 4.9±0.31 and 4.9±0.31 which was significantly higher than the mean antibody titers shown by the PPR (Nigeria 75/1) vaccinated animals i.e. 0±0.00, 3.3±0.51, 3.4±0.51, 4±1.15 and 4.1±1.19 at zero, 7th, 14th, 28th, 48th day post-vaccination respectively. Similarly the mean antibody titers shown by the PPR (Nigeria 75/1) vaccinated animals were 0 ±0.00, 10.4± 3.86, 11.2±4.13, 20±11.31 and 21.6±11.80 at zero, 7th,14th, 28th and 48th day post vaccination respectively. Result of present study demonstrated
Summary
42
that the mean antibody titer values of animals vaccinated with Raksha PPR (Sungri 96) was significantly higher than animals vaccinated with PPR (Nigeria 75/1) at zero, 7th,14th, 28th and 48th day post vaccination respectively. The study also concluded that the mean antibody titer of animals receiving vaccination along with vitamin supplementation was significantly higher than animals receiving only vaccination. While performing the statistical analysis of data, it was revealed that the results were significant (p<0.05).
The present study summarized and concluded that the mean antibody titer values of Raksha PPR (Sungri 96) was significantly higher than PPR vaccine (Nigeria 75/1). As both India and Pakistan are two neighbouring countries, so PPR among them also falls in trans-boundary disease category. It signifies that both being part of Asia subcontinent and PPRV strain of lineage IV prevails in both regions. Keeping these factors under consideration proper vaccination strategy should be followed for the immunization of animals. In past, Nigeria 75/1 strain of PPRV vaccine had been used in Pakistan but the results were not reliable in terms of desired immune response and protection. Although titer was shown by this vaccine but protection is not reliable for proper health care of small ruminants. There was an immense need to come up with the authentic research on PPRV vaccine Raksha PPR (Sungri 96) in Pakistan which is already being used in India with desirable results.
The results of present research project were mostly similar with the findings of other scientists. The results of this study were analyzed through Independent t-test for independent samples. Availability: Items available for loan: UVAS Library [Call number: 2385-T] (1).
20.
Evaluation Of Antiviral Activity And Embryonic Toxicity Of Momordica Charantia Against Newcastle Disease Virus
by Muhammad Usman Ahmed (2013-VA-565) | Dr. Muhammad Adil Rasheed | Dr. Aqeel Javeed | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Plant products play a vital role in the management of various ailments due to their therapeutic activity. A wide range of active phytochemicals peptides have been found to have therapeutic uses against various functionally and genetically diverse viruses. Newcastle disease virus causes respiratory diseases in humans, birds and other mammals, representing one of the foremost threats to public health.
In this study, the antiviral activity of Momordica charantia L. and Ribavirin against Newcastle disease virus was evaluated in-ovo. For each extract of the plant M. charantia and ribavirin 40 embryonated eggs were assigned to 8 groups containing 5 eggs in each group (six groups for antiviral, six groups for embryonic toxicity, and two groups were kept positive and negative control respectively) and marked them with lead pencil. The aqueous and ethanolic extracts of Momordica charantia L. was prepared by using soxhlet extraction technique. From the extract, six different dilutions i.e. 160mg/ml, 80mg/ml, 40mg/ml, 20mg/ml, 10mg/ml and 5mg/ml of the aqueous and ethanolic extracts were prepared in normal saline whereas; six different dilutions i.e. 15μg/ml, 20μg/ml, 25μg/ml, 30μg/ml, 35μg/ml and 40μg/ml of ribavirin were made in normal saline.
With ND virus the different concentrations of the extracts of plant were mixed and 0.2 ml of this suspension was injected to 9th to 10th day embryonated eggs along with positive and negative controls having only virus and normal saline correspondingly. Ribavirin, standard drug, was inoculated by following the mentioned manner. These inoculated embryonated chicken eggs were incubated at 370C and were checked after 12 – 72 hours. After 72 hours of post inoculation, all the eggs were chilled at 40C in fridge for overnight stretch of time and the allantoic fluid was collected.
Summary
64
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 Momordica charnatia aqueous and ethanolic extracts and ribavirin was assessed by merely inoculating the extracts of respective concentrations as used for antiviral activity in embryonated chicken eggs and incubating for 72 hours. The outcomes were analyzed by ANOVA by means of SPSS. Availability: Items available for loan: UVAS Library [Call number: 2384-T] (1).
21.
Detection Of Prevalent Strain Of Ppr Virus And Efficacy Of Imported Live Attenuated Ppr Vaccine In Local Goat In Pakistan
by Iqra Javaid (2008-VA-76) | Prof.Dr. Aneela Zameer Durani | Dr.M. Hassan Saleem | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Peste des petits ruminants (PPR) is a viral, extremely transmissible disease with 100% and 90% of morbidity and death rate in small ruminants (Singh et al. 2004; Singh et al. 2009). The morbillivirus of the family Paramyxoviridae is responsible for its etiology (Barrett et al. 2005). The clinical signs of Peste des petits ruminants (PPR) are severe pyrexia, oculo-nasal discharge, necrotizing and erosive stomatitis, enteritis and pneumonia (Dhar et al. 2002) and is also accompanied by decrease in lymphocyte count (Rajak et al. 2005). Peste des petits ruminants (PPR) produces a major impact on the economy of the country (Zahur et al. 2009). Because of huge economic blow, the Peste des petits ruminants (PPR) imposes a major limitation on sheep and goat production (Asim et al. 2009; Abubakar and Munir. 2014).
The homologous Peste des Petits Ruminants (PPRV) vaccines using Nigeria 75/1 strain of the virus are being manufactured in Pakistan (Asim et al. 2009). The Advance Studies in Vaccinology and Biotechnology Center (CASVAB) University of Baluchistan, Quetta, with the help of Vero cell line manufactured the freeze dried and tissue culture based PPR virus (PPR 75-1) vaccine (Abbas et al. 2011). The homologous and Vero cell based live attenuated PPR vaccine having origin of Indian virus isolate “PPRV-Sungri/96”is being manufactured in India for immunization against Peste des Petits Ruminants (PPR) disease (Sreenivasa et al. 2000).
Twenty goats of different age, breed and sex were examined for the presence of PPR disease during this study. About 2-3 ml of saliva was collected from oral cavity of twenty PPR suspected goats in falcon tubes, signifying PPR disease. The extraction of RNA from the samples was done by trizole method and the concentration was measured by nanodrop. The extracted samples were then subjected to one step RT-PCR and then the PCR products were sent for sequencing to detect the PPRV strain under field conditions.
To study immunogenic behavior of Raksha PPR (Sungri 96), total of forty (40) goats free from peste des petits ruminants virus (PPR-V) were selected for the experimental study. The Group A comprising of twenty (20) goats of age (06 months-01 Year) were further subdivided into two groups i.e subgroup A1 comprising of 10 goats to which Raksha PPR vaccine (Sungri 96) was administered and other ten of sub-group A2 served as control. Similarly the Group B possessing twenty (20) goats of age (01 Year - 02 Year) were further subdivided into two sub-groups i.e subgroup B1 comprising of 10 goats to which Raksha PPR vaccine (Sungri 96) was administered and other ten of sub-group B2 served as control.
The RNA concentration was different in all twenty saliva samples when measured by nanodrop. Only five (5) samples out of total twenty (20), saliva samples from PPR suspected goats, were positive through RT-PCR and yielded an amplified product of 351bp. The five amplicons were sent for sequencing and the phylogenetic tree was constructed. The tree demonstrated that the Pakistani strains of PPRV clustered into lineage IV showing similarity with the isolates from China, Kurdistan, Iran and Bangladesh.
It was revealed that the that the animals (1 year to 2year old ) vaccinated with Raksha PPR (Sungri 96) displayed the significantly higher mean antibody titers than the mean antibody titers shown by vaccinated animals of age (6 months to 1 year) at zero, 7th, 14th, 28th, 48th day post vaccination respectively. On statistical analysis of data, the results were significant (p<0.05).
The present study revealed the presence of lineage IV in Pakistan. This will help to plan proper control strategies against this deadly viral disease. Currently the Nigeria75/1 vaccine is being used in Pakistan which clusters in lineage II while Pakistani field isolates fall under lineage IV. So it is very important to immunize the animals with lineage specific vaccine like Raksha PPR (Sungri 96) manufactured by IVRI, India.
This study reported the strong association of age and PPR vaccination titer in goats. Our findings concluded that the strong immune response was shown by adult animals against PPRV vaccine as compared to young stock. The results of present research project were mostly similar with the findings of other scientists. The results of this study were analyzed by one way ANOVA for independent samples.
Availability: Items available for loan: UVAS Library [Call number: 2381-T] (1).
22.
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).
23.
In Vitro Antibacterial Evaluation Of Ceftriaxone Alone And In Combination With Ascorbic Acid In Post Surgical Infections
by Umbreen Anwar (2013-VA-853) | Dr. Muhammad Adil Rasheed | Dr. Muhammmad Ovais Omer | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Nosocomial infectionshave frequently been reported by several hospitals worldwide. A patient infected with such infections is presented with complaint of fever, inflammation, redness and pus. These infections are also called post-surgical or surgical site infections and a suitable antibiotic therapy can be used to cure these infections(Soriano et al. 2006).Nosocomial infections occur in patients with in 2 days of admittance in hospital three days of liberation or 30 days of incision, it causes increase in hospital stay in infected patients (Inweregbu et al.2013). Surgical site infections including urinary tract infection and pneumonia are most common hospital acquired infections and are caused by bacteria, viruses and fungus (Timsitet al.2012).
These infections occur up to thirty days after surgery (Owenset al.2008). Surgical Site infections increase the rate of morbidity and mortality among surgically operated individuals (Powell et al. 2005).Controllingfactors for these infections includesage, gender, prophylactic administration of antibiotics and aseptic procedures(Lavallee et al. 2014).
There are number of risk factors in development of hospoital acquired infections. Obesity is also an important risk factor in developing post operative infections and is directly associated with under dosage of antibiotic given prophylactically (Hunttunen et al.2013). Surgeon should be aware of antibiotic choice, dose and duration based on reliable guidelines for prophylaxis to avoid common type of adverse effects on surgical sites (Rafati et al. 2014).
Motie et al. (2014) found that there was an inverse relation between length of surgical incision and rate of surgical infections it was found that type of surgery is main risk factor in developing of infections. The most commonly prescribed antibiotics were combination of ceftriaxone and metronidazole (51.6%).The contaminated and clean contaminated wounds are associated with higher rate of surgical site infections.
Post-surgical infections are known to major health issue, that are responsible for high treatment cost, more readmission in hospitals, increased stay in hospital and increase in rate of infections and even death of surgically operated patient (Mengesha et al. 2014). The rate of occurrence increases due to use of mini sterile gloves, operating costumes, face masks and other specific surgical coverings in operating rooms (Salassa and Swiontkowski 2014).
Both Gram positive and Gram negative species of bacteria are responsible in causing such infections and most common isolates obtained from pus areStaphylococcusepidermidis, Staphylococcus aureus, Proteus vulgaris,Pseudomonas aeruginosa, Klebsiella pneumonia, Proteus mirabillis, Escherichia coli etc.Post surgical abdominal infections are more common after abdominal surgeries. But the disclosure of these infections in early stage is considerably difficult in old age patients along the condition of hyperthermia and increased level of C reactive protein are visible sign in such type of infections (Lin et al. 2002).A variety of natural and synthetic antibiotics are used to treat infections. Ascorbic acid, zinc and garlic are useful in killing bacteria, improving immunity and thus preventing diseases.The most important thing about these antibiotics is that these are less toxic and less harmful thus can be used in pregnant women reducing the chance of urinary tract infections. Also the intake of 100mg ascorbic acid or vitamin c as important vitamin in balanced diet daily causes improvement of health in pregnant women(Ochoa et al. 2007).
There is much rapid increase in morbidity and mortality in postpartum women day by day although many antibiotics are used peri operatively along with other precautions and preventive measures. So obstetricians are facing a lot of complications in operatingmany caesarean deliveries (Haas et al. 2014).
The usage of appropriate antibiotics peri operatively for prophylaxis along following withan appropriate aseptic procedure before, during and after surgery proved effective in controlling these infections (Michalopoulos and Sparos 2002).
Optimal dosage and duration while administration of prophylactic antibiotics to patients must be considered because use of antibiotic for inappropriate duration of time can result in increased risk of post-operative brain infections (Wu et al. 2013).A large number of cephalosporins speciallyof second generationhave been proved very effective in reduction of development of post-operative infections but dosage and duration of these antibiotics varied from patient to patient (Gelijns et al. 2014).Intra-abdominal pus can be reduced by the correct use of antibiotics and by following sterile procedures (Romano et al. 2014).Cephalosporins are useful for eradication of pathogens of skin like Staphylococcus aureus. An antibiotic cefazolin belonging to first generation antibiotics plays major role against pathogens in many clean wound incisions (Page et al.1993).
In field of gynaecology surgery some antibiotics are also prescribed in combinations and thus proved more efficacious (Bratzler et al. 2013). Ceftriaxone from the group of third generation cephalosporins given before surgery was as useful in prevention of major pelvic infections and urinary tract infections as compared to three doses of cefazolin given over 14 hours peri operatively (Hemsell et al. 1984).
Ascorbic acid is a potentantioxidant which markedly reduced the growth of E.coli, Pseudomonas aeruginosa and Staphylococcusand it provided significant effectiveness in combination with levofloxacin (Carlssonet al.2005).Ascorbic acid increases wound healing, immune system activation, collagen formation due to its oxidative property.Alsonutritional deficiencies decreases wound healing after surgeries (MacKay and Miller 2003).Ascorbic acid inhibits the growth of Stayphylococusaureus bacteria by producing oxidative radical thus increasing the oxidative stress,affecting the metabolism and inhibiting the growth of bacteria invitro(Kallio et al. 2012).
The use of ascorbic acid with antibiotics is significant, high doses of ascorbic acid with antibiotics have shown synergistic effects and resulted in prevention of life threating diseases thus high potency vitamin supplementation can reduce morbidity and speed recovery (Ishida et al.1998).
The aim of this study project is to identify the prevalence of microbes involved inpost-operative infections and also to determine the sensitivity patterns of isolated pathogens by using culture sensitivity test against most commonly prescribed antibiotic (ceftriaxone) alone and in combination with ascorbic acid (vitamin C) in vitro.
Availability: Items available for loan: UVAS Library [Call number: 2466-T] (1).
24.
Antibacterial Activity Of Piroxicam And Ketorolac Alone And In Combination With Antibiotics Against Bacterial Isolates
by Saba Shahbaz (2013-VA-852) | Dr. Muhammad Ovais Omer | Dr. Muhammad Adil Rasheed | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Antibiotic resistance has become a global public health problem due to the excessive and indiscriminate use of antibiotics, which has resulted in many emerging multidrug-resistant microorganisms. This study is designed for the evaluation of different dilutions of piroxicam and ketorolac alone and in combination with amoxicillin and tigecycline by using broth dilution method.Different dilutions of piroxicam and ketorolac alone and in combination with amoxicillin and tigecycline were checked for antibacterial activity against Staphylococcusaureus and Escherichia coli.The isolates were obtained from Quality Operations Laboratory. The pathogens were tested for their sensitivity to amoxicillin and tigecycline. The sensitivity was checked by broth/tube dilution method. Dilutions were prepared by two fold dilution method.Collected data was analyzed by using statistic package for social sciences (SPSS, windows version, Chicago, IL, USA). Analysis of Variance (ANOVA) and descriptive statistics was applied.This work is designed to observe the effects of piroxicam and ketorolac alone and in combination with amoxicillin and tigecycline against bacterial pathogens to improve the quality of life of patients and will minimize the chances of infections.Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) was determined by broth dilution method. The results suggested that NSAIDs enhanced the antibacterial effect when combined with these antibiotics. Combination of amoxicillin with piroxicam (9.76µg/ml+80µg/ml), and tigecycline in combination with ketorolac (0.156µg/ml+20µg/ml) was effective against Staphylococcusaureus. The combination of amoxicillin with piroxicam (9.76µg/ml+20µg/ml), amoxicillin with ketorolac (4.88µg/ml+20µg/ml), tigecycline with piroxicam (0.3125µg/ml +10µg/ml), tigecycline with ketorolac (0.312µg/ml+20µg/ml) showed efficacy against Escherichia coli. Availability: Items available for loan: UVAS Library [Call number: 2448-T] (1).
25.
Effect Of Stabilizers On Biological Titre Of Freeze Dried Ppr Virus Vaccine
by Muhammad Zubair Latif (2009-VA-382) | Prof. Dr. Khushi Muhammad | Dr. Muhammad Anees | Dr. Imran Altaf | Dr. Muhammad Imran.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: PPR is an acute and highly contagious viral disease of small ruminants caused by Morbillivirus. It causes high morbidity and mortality in small ruminants and heavy economic loses to farmers. Live attenuated vaccines are commonly used to control the disease. During freeze drying and after dilution of freeze dried vaccine, there is lose of virus titre so vaccine efficacy is reduced. Stabilizers are therefore added to protect from freeze drying stress and heat shock during storage and transportation. Each stabilizer has different protective effect. The present project was therefore designed to evaluate different stabilizers to act as the best one for maximum stability of the virus titre.
Fifty vials of PPR vaccine with each of six stabilizers (Weybridge medium-WBM, Lactalbumin hydrolysate sucrose-LS, lactalbumin hydrolysate sorbitol-LSbG, Tris sucrose-TS, Tris Trehalose-TT and Goat skimmed milk-GSM) was formulated, freeze dried and three vials from each formulation was selected and evaluated by biological titration just after freeze drying and dilution in PBS (7 pH). Each of the vaccine diluted with the PBS and stored at 40C, 250C, 370C for 36 hours. Biological titration of each of the vaccines stored at different temperature and time was determined on Vero cell lines. The virus infectivity was calculated as mean TCID50 by MTT assay.
It is concluded from the study that stabilizers having carbohydrates (sucrose, sorbitol, trehalose), salts (sodium) and hydrolyzed proteins (lactalbumin hydrolysates) are effective to make compact mass (freeze drying) of PPR virus vaccine. WBM, LS, and LSbG maintain infectivity of the PPR virus vaccine (if reconstitution with PBS and stored at 4°C) for 12 hours. However, TT is able to protect infectivity titre of the PPR virus vaccine during freeze drying and even during its storage after hydration with PBS at 4°C for 24 hours. Availability: Items available for loan: UVAS Library [Call number: 2546-T] (1).
26.
Comparative Quality Evaluation Of Raw And Pasteurized Milk
by Hafiza Saima Ghaffar (2009-VA-230) | Dr. Imran Altaf | Prof. Dr. Aftab Ahmed Anjum | Dr. Sana Ullah Iqbal.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: This particular project was designed to evaluate the overall quality of raw and pasteurized milk available at different areas of Lahore. The parameter which was checked includes microbiological analysis, adulterants, physicochemical properties and the effect of temperature on vitamin C in milk samples. Raw samples were collected from ten different towns of Lahore, whereas pasteurized milk samples belong to ten different brands. Ten samples were collected under control conditions from animals in sterilized containers.
For microbiological analysis four parameters were selected including total plate count (TPC), total coliform count (TCC), total psychrotrophic count (TPSC) and total yeast and mold count (TYMC) whereas, different adulterants like adulteration test was done such as urea, starch, hydrogen peroxide, detergent or soap, sorbitol, quaternary ammonium compound, boric acid, cane sugar, sodium chloride, formalin and hypochlorite were checked by using the milk adulteration kit in QOL. Milk contains casein and whey proteins. Whey protein being added in the milk to increase its density which is considers being an adulterant. In this project whey protein was estimated in all milk samples by titration method. Physicochemical characteristics of milk are an important parameter to judge the quality of milk. These physicochemical properties include fat%, SNF%, density kg/m3, lactose%, solid/ash, protein% and pH. Physicochemical properties were evaluated mechanically by Milkoscan. Heat treatment is an important method to reduce the microbiological contamination of milk. These treatments may include pasteurization and UHT etc. During the heat treatment some of the micronutrients may deteriorate thus compromising the quality of milk. Vitamin C is among those heat labile micronutrients. Vitamin C was checked quantitatively in market and self-collected samples by using titration method.
It was concluded that total plate count TPC, TCC, TPSC and TYMC of raw milk samples were above the standard value indicating the poor quality of the milk. As far as the pasteurized milk samples were concerned ninety percent of the samples showing higher values for TCC, TPSC and TYMC. Total plate counts of all self-collected raw milk from a healthy animal were found within the standard value. Counts were in range of 3.8x 103 – 8.9x103 CFU/mL of all milk samples. TPC of all self-collected raw milk from a healthy animal were found within the standard TCC were found within permissible value (102 CFU/mL .TPSC were negative for all milk samples. TYMC were in range of 2.6x101 -7.2x101 CFU/mL. Among milk samples (n=10), three samples (30%) were positive for TYMC were while remaining samples (70%) were negative and showed no growth.
Physicochemical factor show that 50 percent of raw milk have low nutritional value as compared to the standards which are buffalo and cow milk contains 7.6, 4.5% fat, 3.8, 3.8 % protein, 5.1, 4.9% lactose, 0.78, 0.72% ash and 17.0, 13.9% total solid respectively. In raw milk mean of fat (%), solid not fat (%), lactose (%), Solid/ash (0%), protein(%) and pH were 4.50±0.03, 7.915±0.06, 23.05±0.055, 3.893±0.06, 3.85±0.05, and6.9±0.0.02 respectively. In pasteurized milk mean value for fat, SNF, lactose, ash, protein and pH were 3.48 ±0.13, 7.24±0.10, 3.60±0.05,0.5 ±0.06, 2.82±0.05, 7.2±0.20 respectively. Pasteurized milk is good for consumption.
Different adulterant such as urea, starch, hydrogen peroxide, Sorbitol, QAC, Boric acid, Cane sugar, NaCl, Carbonate, Formalin, hypochlorite, whey protein, Added water and soap /detergents were evaluated in all milk samples. Among these adulterant water (66%) was found in majority of milk samples, followed by whey protein (15%), starch (13%), (10%) NaCl and
(8%) cane sugar were detected in raw milk samples. n Pasteurized milk samples only added water (49%) and whey protein (31%) was detected.
Among the raw milk samples the maximum and minimum concentration of vitamin C was observed 0.33±0.02 and 3.33 ±0.02 mg/100ml and for pasteurized milk maximum and minimum concentration of vitamin C was observed 2.54mg/100ml and 0.32 ±0.02 mg/100ml respectively. In self- collected samples the minimum and maximum concentration of vitamin C was observed 5.25±0.02 and 8.34 ±0.04 mg/100ml respectively and after pasteurization in laboratory minimum and maximum concentration of vitamin C was observed 3.48±0.04 and 5.83 ±0.02 mg/100ml respectively. These observations had showed that pasteurization treatment decreased Vitamin C quantity. Availability: Items available for loan: UVAS Library [Call number: 2536-T] (1).
27.
Evaluation Of Antiviral Activity And Embryonic Toxicity Of Ivermectin And Ibuprofen Alone And In Combination Against Avian Influenza H9
by Huma Minhas (2014-VA-499) | Dr. Muhammad Ovais Omer | Dr. Qamar Niaz | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: This project was designed to analyze the antiviral andembryotoxicity of ivermectin and ibuprofenalone and in combination against H9 virus by using embryonated chicken eggsof 10 days old. Three different concentrations of these agents were selected for current project and two fold dilutions were made. Mixing of drug dilutionnswith avian influenza H9 virus was done and administered in embryonated eggs. They were kept in incubator for 72 hours. Eggs viability was checked during incubation at 37°c temperature. After overnight chilling,haemagglutinition test was done to evaluate antiviral activity. Antiviral activity of these dilutions was calculated as embryo survival percentage and positive and negative hemagglutination activity. To checkembryotoxicity,drug dilutions were made without virus and mortality ratio was checked after 48 hours of incubation.
The study provided information regarding antiviral activity and embryotoxicity of ivermectin and ibuprofen alone and incombination at different concentrations. The present study showed that antiviral activity of ivermectin was very strong at all concentrations however at higher concentration it was toxic for embryo.
Results of antiviral analysis showed that ivermectinand ibuprofen had antiviral activity alone and in combination afterusing combination of ivermectin and ibuprofen the antiviral activity was further increased and embrytoxicity was also diminished by combination therapyso 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 ivermectin and ibuprofen alone and in combination showed that ivermectin had very strong antiviral activity but it was embryotoxic at higher concentrations when ibuprofen was used in combination then had further strong antiviral activity. It’s antiviral activity was stronger as compare when these agents used alone. In term of embryotoxicity these these agents were not toxic in combination.
Availability: Items available for loan: UVAS Library [Call number: 2588-T] (1).
28.
Affect Of Temperature, Cell Density And Multiplicity Of Infection On Biological Titer Of FMDV Type “O”
by Qazi Ithram Ul Haq (2009-VA-104) | Dr. Imran Altaf | Prof. Dr. Aftab Ahmad Anjum | Dr. Muhammad Imran.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: FMD is a transmissible viral disease of animals. It is causing very highly economical loses in Pakistan and all over the world. Through vaccination FMD is being controlled in Pakistan. Inactivated virus is used in vaccines. FMD virus grows on BHK-21 cell lines. FMDV show good adoptability on these cell lines. For good and high titer FMDV needs few physical factors to grow on BHK-21 cell lines. These factors include Temperature, Cell density and Multiplicity of infection (MOI)was considered in this research. The FMDV strain “O” was grown on BHK-21 cell line. The cells monolayer was propagated for conduction of effect of these factors on the virus. The mentioned factors were studied to get optimum level of virus titer in in vitro cell lines. The effect of 35°C, 37°C and 39°C was evaluated on the virus growth. Maximum virus propagation was noted at optimum temperature 37°C. The viral concentration at 37°C was significantly (P<0.05) higher than at 35°C and 39°C. The effect of cell density was studied on the virus concentration. Flask of three different densities 25cm2, 175cm2 and 275cm2were utilized in the current study. The virus concentration in all three different densities was not significantly different (P>0.05) from each other. Another factor Multiplicity of infection (MOI)was investigated in the study. Five different volumes 10ul, 20ul, 30ul, 40ul and 50ul of the FMDV strain “O” were used to investigate the effect of factor on the virus concentration. The results revealed highest viral harvest concretion at 50ul volume with MOI of 7.1, %age of cells infected with single virus and 6.3 × 1079. The MOI at 50ul was significantly higher (P<0.05) than the other four concentration of the virus. It was concluded from the study that the optimum temperature for the maximum FMDV concentration harvest is 37°C. The density of cell has no significant effect on the growth of virus that is flask of any density may be used to grow the FMDV. Multiplicity of infection (MOI) of 14.2 give maximum
SUMMARY
34
TCID50 Optimizing the conditions for the cell culture and virus cultivation helps in maximum virus harvest achievement. From the present study it may be suggested that the physical factors may be optimized for the remaining strains of FMD and other vaccine viruses to attain maximum virus grow. Availability: Items available for loan: UVAS Library [Call number: 2681-T] (1).
29.
A Comparative Study Of Non-Antibiotic Feed Additives On Experimental Colonization Of Salmonella Enterica Serovar Enteridis And Intestinal Pathomorphology In Broiler Chickens
by Adeem Rehman Raffie (2010-VA-233) | Prof. Dr. Asim Aslam | Dr. Muhammad Yasin Tipu | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: The utility of antimicrobial agents as a preventive measure has been questioned, given extensive documentation of the evolution of antimicrobial resistance among pathogenic bacteria. Non-antibiotic feed additives (probiotics, prebiotics, essential oils and organic acids) are being considered to fill this gap and already a few farmers in the country are using them with good results. The present study enable us to understand and compare the beneficial effects of non-antibiotic feed additives on salmonella enterica colonization and changes in intestinal morphology.
This study is designed to evaluate the effect of non-antibiotic feed additives on salmonella enterica serovar enteritidis colonization in intestine of broilers chickens and compare the intestinal morphology between normal healthy, non-antibiotic feed additives supplemented and salmonella challenged broiler chickens.
A total of 125 commercial day-old broiler chicks were procured from the local market. The chicks were divided into six groups A (Basal diet, negative control group), B (Challenge + Basal diet, positive control group), C (probiotic + Challenge + Basal diet), D (prebiotic + Challenge + Basal diet), E (essential oils + Challenge + Basal diet) and F (organic acids + Challenge + Basal diet) with 20 chicks in each group and given separate treatments.
Two separate experiments were carried out for salmonella recovery from cecal tonsils and intestinal pathomorphic evaluation. Villus length, villus width, villus surface area and crypt depth were measured by micrometery. The collected data from both experiments was analyzed using the statistical technique of comparing more than two groups i.e. Analysis of variance (ANOVA) through SPSS 16.0.
Summary
45
There was an overall increase in all the parameters of intestinal morphometric analysis for all the treatment groups except for the control negative group which showed lowest values. Maximum villus height of 1794.2±63.96 μm in duodenum was achieved by group E, which was given essential oils. Whereas maximum villus surface area index of 1662.6±389.16 mm2 was recorded in group D, which was treated with prebiotics. Maximum villus height of 940.35±23.96 μm and surface area index of 568.92±36.27 mm2 in ileum mucosa was recorded in group D, treated with prebiotic. . Recoverable salmonella was most reduced by probiotics and organic acids.
Final results show that there is an overall increase in histological parameters of the mucosa of duodenum and ileum in the groups fed non-antibiotics feed additives as compared with control groups. Prebiotics showed the maximum positive effects in histological parameters whereas probiotics showed maximum positive effect for decreased recoverable salmonella count. Hence this study suggests that a combination of non-antibiotic feed additives will be beneficial for the intestinal health of broiler chickens but there is a need for more research on combinations of non-antibiotic feed additives. Availability: Items available for loan: UVAS Library [Call number: 2844-T] (1).
