51.
Molecular Identification And Treatment Of Theileriosis In Small Ruminants Of Northern Balochistan
by Mir Ahmad Khan (2005-VA-214) | Prof. Dr. Muhammad Arif Khan | Prof. Dr. Muhammad Azam Kakar | Prof. Dr. Muhammad Sarwar Khan | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The present study was conducted to investigate the prevalence of Ovine and Caprine Theileriosis in Northern Highlands and Suleiman Mountain Region of Balochistan, Six thickly populated /union councils were included in the study area. Samples were collected from 2870 animals Sheep (n= 2200) and Goats (n= 670) for screening of the disease. The samples were collected and processed in Regional Disease Investigation Laboratories, Department of Livestock and Dairy Development Balochistan, T.B. Sanatorium Hospital Quetta and Center for Vaccinology, Bacteriology, The University of Balochistan, Quetta and Medicine Laboratory, Department of Clinical Medicine and Surgery, The University of Veterinary and Animal Sciences, Lahore. Data revealed 20.82% disease in sheep and 9.70%. in goats. The regional prevalence of theileriosis revealed 19.19% in Northern Highlands and 17.48% in Suleiman Mountain Region Chi-square analysis showed significant difference in the prevalence of disease in sheep and goats. The regional difference was not significantly different between two regions of Northern Balochistan. The comparison among union councils showed significant difference being highest prevalence (22.71%) in union council Kuchlak district Quetta followed by Aghberg (18.42%) and Hanna Urak (15.53%) in Northern highlands and Union Council Zangiwal Jogezai (19.83%) followed by Kach Amaqzai (16.30%) and Sinjavi (15.92%) in SMR. The disease prevalence when compared among 4 different breeds of sheep showed significant difference being highest in Karakul breed (34.62%) followed by Shinwari (24.54%), Bibrik (19.36%) and Harnai (16.40%). The highest prevalence of theileriosis in sheep and goats were observed in Summer season (30.30%) followed by Autumn 19.07%, Spring 14.52% and Winter
SUMMERY
105
7.61%. Chi-square analysis of the data showed significant difference in the prevalence of the disease in different seasons of the year. The disease was also compared in three age groups of sheep and goats. The data showed 22.17% disease in adult animal group above 2 years of age followed by 15.85% in animals between 1-2 year and 7.99% in age group below one year. Statistically significant difference in all age groups was found in chi-square analysis. The sex wise prevalence of theileriosis revealed non-significant difference between male and female sheep and goats. Two different species of Theileria were reported by many researchers causing disease in sheep and goats. The PCR was carried out for the identification of Theileria species affecting sheep and goats in Balochistan. Two species specific sets of primers were designed using 18SRNA gene sequence to identify these two species of Theileria and the distribution among the two species of animals. The genomic DNA of two species of parasite was successfully amplified in positive samples. The assay was proved successful and we recommend for the prevalence surveys for theileriosis in sheep and goats. The data showed that the prevalence of T. lestoquardi was 73.80% in sheep and 69.23% was in goats in the target regions. It was found the T. lestoquardi was highly prevalent and causing theileriosis in small ruminants. The prevalence of T. ovis was 26.19% in sheep and 30.76% in goats respectively in the investigated animals; it was less than T. lestoquardi. It was concluded that both Theileria species were identified and found circulating in small ruminants in the target region of Balochistan. In the study we determined that PCR method based on 18S RNA gene could detect and differentiate T. ovis and T. lestoquardi.
Effect of theileriosis in sheep and goats on hemeto-biochemical parameters were studied included RBCs, Hb%, PCV, Platelets, WBCs, MCV, MCHC, AST, ALT, BUN, Bilirubin and Creatinine. Blood samples were collected from Theileria confirmed, diseased animals (sheep and
SUMMERY
106
goats) along with equal number of healthy animals for comparison. In sheep RBCs, Hb%, PCV, WBCs, MCHC, AST, ALT and Creatinine values showed significant difference when compared with values of healthy animals. Significant (p<0.05) reduction was noted in measurement of RBCs, Hb%, PCV and MCHC whereas, AST, ALT and Creatinine showed significant increase in diseased animals. In goats affected with theileriosis showed significant decrease in RBCs count and Hb%. The values for AST, ALT and Creatinine were found significantly increased in diseased animals when compared with healthy control group of equal number of animals. In present study it was noted that Butalex intra muscularly at the rate of 2.5 mg/kg body weight is quite effective in eliminating the Theileria parasite from the blood of sheep and goats and treatment at the day 10 post treatment. Imizol was also found an effective treatment of theileriosis but less effective than Butalex. Availability: Items available for loan: UVAS Library [Call number: 2690-T] (1).
52.
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).
53.
Molecular Identification And Treatment Of Theileriosis In Small Ruminants Of Northern Balochistan
by Mir Ahmad Khan (2005-VA-214) | Prof. Dr. Muhammad Arif Khan | Prof. Dr. Muhammad Azam Kakar | Prof. Dr. Muhammad Sarwar Khan | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The present study was conducted to investigate the prevalence of Ovine and Caprine Theileriosis in Northern Highlands and Suleiman Mountain Region of Balochistan, Six thickly populated /union councils were included in the study area. Samples were collected from 2870 animals Sheep (n= 2200) and Goats (n= 670) for screening of the disease. The samples were collected and processed in Regional Disease Investigation Laboratories, Department of Livestock and Dairy Development Balochistan, T.B. Sanatorium Hospital Quetta and Center for Vaccinology, Bacteriology, The University of Balochistan, Quetta and Medicine Laboratory, Department of Clinical Medicine and Surgery, The University of Veterinary and Animal Sciences, Lahore. Data revealed 20.82% disease in sheep and 9.70%. in goats. The regional prevalence of theileriosis revealed 19.19% in Northern Highlands and 17.48% in Suleiman Mountain Region Chi-square analysis showed significant difference in the prevalence of disease in sheep and goats. The regional difference was not significantly different between two regions of Northern Balochistan. The comparison among union councils showed significant difference being highest prevalence (22.71%) in union council Kuchlak district Quetta followed by Aghberg (18.42%) and Hanna Urak (15.53%) in Northern highlands and Union Council Zangiwal Jogezai (19.83%) followed by Kach Amaqzai (16.30%) and Sinjavi (15.92%) in SMR. The disease prevalence when compared among 4 different breeds of sheep showed significant difference being highest in Karakul breed (34.62%) followed by Shinwari (24.54%), Bibrik (19.36%) and Harnai (16.40%). The highest prevalence of theileriosis in sheep and goats were observed in Summer season (30.30%) followed by Autumn 19.07%, Spring 14.52% and Winter
SUMMERY
105
7.61%. Chi-square analysis of the data showed significant difference in the prevalence of the disease in different seasons of the year. The disease was also compared in three age groups of sheep and goats. The data showed 22.17% disease in adult animal group above 2 years of age followed by 15.85% in animals between 1-2 year and 7.99% in age group below one year. Statistically significant difference in all age groups was found in chi-square analysis. The sex wise prevalence of theileriosis revealed non-significant difference between male and female sheep and goats. Two different species of Theileria were reported by many researchers causing disease in sheep and goats. The PCR was carried out for the identification of Theileria species affecting sheep and goats in Balochistan. Two species specific sets of primers were designed using 18SRNA gene sequence to identify these two species of Theileria and the distribution among the two species of animals. The genomic DNA of two species of parasite was successfully amplified in positive samples. The assay was proved successful and we recommend for the prevalence surveys for theileriosis in sheep and goats. The data showed that the prevalence of T. lestoquardi was 73.80% in sheep and 69.23% was in goats in the target regions. It was found the T. lestoquardi was highly prevalent and causing theileriosis in small ruminants. The prevalence of T. ovis was 26.19% in sheep and 30.76% in goats respectively in the investigated animals; it was less than T. lestoquardi. It was concluded that both Theileria species were identified and found circulating in small ruminants in the target region of Balochistan. In the study we determined that PCR method based on 18S RNA gene could detect and differentiate T. ovis and T. lestoquardi.
Effect of theileriosis in sheep and goats on hemeto-biochemical parameters were studied included RBCs, Hb%, PCV, Platelets, WBCs, MCV, MCHC, AST, ALT, BUN, Bilirubin and Creatinine. Blood samples were collected from Theileria confirmed, diseased animals (sheep and
SUMMERY
106
goats) along with equal number of healthy animals for comparison. In sheep RBCs, Hb%, PCV, WBCs, MCHC, AST, ALT and Creatinine values showed significant difference when compared with values of healthy animals. Significant (p<0.05) reduction was noted in measurement of RBCs, Hb%, PCV and MCHC whereas, AST, ALT and Creatinine showed significant increase in diseased animals. In goats affected with theileriosis showed significant decrease in RBCs count and Hb%. The values for AST, ALT and Creatinine were found significantly increased in diseased animals when compared with healthy control group of equal number of animals. In present study it was noted that Butalex intra muscularly at the rate of 2.5 mg/kg body weight is quite effective in eliminating the Theileria parasite from the blood of sheep and goats and treatment at the day 10 post treatment. Imizol was also found an effective treatment of theileriosis but less effective than Butalex. Availability: No items available
54.
In Vitro Activity Of Selected Biocides Against Fungal Isolates From Production Area Of Pharmaceutical Industry
by Sana Ilyas (2009-VA-238) | Dr. Muhammad Nawaz | Prof. Dr. Aftab Ahmad Anjum | Dr. Muhammad Ovais Omer.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Pakistan pharmaceutical industries have grown to grab their position amongst top ten pharmaceutical industries of Asia Pacific region. These are serving with 80% of pharmaceutical needs. The industry on the other hand faces some challenges in terms of sterile pharmaceutical product manufacturing. The fungal contamination causes spoilage to pharmaceutical products, cosmetics, and food products. The fungal contamination to pharmaceutical products has resulted in direct losses to human health and to economy.
A total of 50 air samples were collected from clean area of a pharmaceutical production unit by exposing sabouraud dextrose agar (SDA) plates by settle plate method (4 hours exposure). Fungal colonies were purified by sub-culturing and later identified macroscopically and microscopically. Selected biocides included isopropyl alcohol (70%), chloroxylenol (20%), chlorhexidine gluconate (20%), and benzalkonium chloride (20%) were used in this study. A 100 μl of spore suspension of each fungal contaminant (1.0 × 106 to 5.0 × 106 spores/mL) was exposed to 9.9 mL of biocide preparation for 15 and 30 minutes while exposure was stopped by adding 1 mL of mixture (spores exposed to biocide) into 9 mL of respective neutralizing agents The enumeration of colonies was started immediately after the growth was visible and expressed as Mean±S.D. and converted to log10. Antifungal activity of biocides was expressed as log10 reduction and different biocides‟ activity was compared using ANOVA technique by graphed prism 5.0 statistical software.
Total 204 colony forming units (CFU) were identified from filling area (36), solution room (47), and buffers (121). The antifungal activity in terms of log reduction was lowest by isopropyl alcohol at 15 minutes and highest was shown by chlorohexidine gluconate at 30 minutes against
Summary
64
Aspergillus flavus. In case of Aspergillus fumigatus all the biocides presented significant difference of antifungal activity at 15 minutes. The response of Aspergillus niger against different biocides at 15 minutes and 30 minutes was same as was in case of Aspergillus flavus while each biocide‟s antifungal activity was found significantly increased with increase in time of exposure. The similar response of antifungal activity of different biocides at both exposure times was noted against Saccharomyces cerevisiae. The antifungal activity of all biocides against penicillium was found significant different at 15 minutes and 30 minutes exposure time. Similarly, each biocide‟s antifungal activity increased with increase in time of exposure. On overall basis, isopropyl alcohol was found less effective while benzalkonium chloride and chlorohexidine gluconate presented comparatively higher efficacy against fungal isolates. Availability: Items available for loan: UVAS Library [Call number: 2705-T] (1).
55.
Study On The Pathogenesis Of Field Isolate Of Salmonella Pullorum In Experimentally Infected Broiler Chicken
by Muhammad Zeeshan (2014-VA-535) | Dr. Muti-ur-Rehman Khan | Dr. Gulbeena Saleem | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Poultry is one of the well organized sectors in Pakistan and generates employment for up
to 1.5 million people. As use of modern techniques and medicine has reduced a risk lot but it is
still facing threat from many diseases. One of these is Salmonellosis which is and hatchery borne
infection and it also has a zoonotic importance. This organism is gram negative bacterium, non
acid fast, uniform staining and non spore forming bacillus.
Pullorum disease produced by it is also known as bacillary white diarrhea due to whitish
droppings produced by birds. Day old chicks are affected because it can be transmitted vertically.
Infected fomites and utensils also play a role in the transmission of infection. Growth of S.
pullorum rapidly occurs upon beef agar on nutrient media and growth is best at 37 °C. Selective
media such as selenite, tetrathionate broths and differential media such as MacConkey, bismuth
sulfide and brilliant green agar are best for the proper growth of organism.
The organism was isolated from the 30 different infected samples of broiler birds taken
from different farms and was grown upon the selective media. The colonies were picked from
the cultures and were subjected to PCR for the further process. After confirmation with PCR,
inoculums was prepared with a bacterial load of 2 x 107 (CFU) in 0.5 ml of normal saline at the
ph of 7.2.
Chicks were randomly divided in to three groups as group 1, 2 and 3, each group
containing 30 chicks. Group 1 was infected with field strain of S.pullorum, Group 2 with
vaccinal strain and Group 3 was set as control group and no infection was given to it. Samples
were collected from birds at 10th, 15th, 20th and 25th days of age. Observations were made upon,
clinical signs, histopathological lesions, mortality and morbidity rate, FCR and gross
pathological lesions.
Clinical signs included were anorexia, depression, huddling, labored breathing, loss of
feed and water intake, ruffled feathers, reduced mean body weights, low FCR and pasty vent.
Gross and histopathological lesions were much observed and were prominent in the birds
affected with field strain of S. pullorum. One Way-ANOVA was used for statistical analysis in
between the groups. Gross pathological lesions included were liver congestion 55%, cheesy
material in caeca 70% in the birds which were affected with field strain of S. pullorum.
Histopathological findings in the birds affected with field strain were monocytes in spleen,
leukocytes in intestine and heterophils in liver. Low FCR was recorded in infected birds of
Group 1 and was 1.63 at 25th day. Mortality rate in the birds affected with field strain was
maximum 43.33% and morbidity rate was 73.33% at the 10th day of age. Birds of group 2 were
less affected and only showed medium level of lesions. No specific lesions were seen in group 3.
The total prevalence of of S.pullorum after conformation with PCR was 20% out of total 30
infected samples. The organs for histopathology taken were liver, spleen, caeca and intestine and
were preserved in 10% formalin prior to the infection.
The overall pathogenesis of experimental bacteria after given through oral route was that
it first localizes in digestive tract from where it enters into the blood stream and invades in the
different organs and tissues at different time intervals producing lesions and immunological
response. Availability: Items available for loan: UVAS Library [Call number: 2693-T] (1).
56.
Microbiological Analysis Of Food Contact Surfaces Used In Various University Cafeterias Of Lahore
by Rabab Akhtar (2014-VA-958) | Dr. Zubair Farooq | Dr. Sana Ullah Iqbal | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Personal hygiene and food contact surfaces are direct routes of cross contamination that can make food unsafe and are a major concern for food service facilities in controlling the spread of food borne pathogens. Good cleaning practices are particularly important in premises like cafeterias, preparing ready-to-eat (RTE) foods, as these foods are consumed without further cooking or processing to reduce or eliminate microorganisms. Lahore is educational capital of Pakistan where educational institutions are increasing and therefore number of students, eating meals from university cafeteria is becoming more frequent. Despite the growth of this sector, there is no effective education or training of the food handlers or hygienic control of the food sold as many of these were sealed last year due to unhygienic conditions. This study aims to evaluate the microbial status of various food contact surfaces that are used in various university cafeterias of Lahore so that recommendations could be provided and therefore risk of food poisoning cases among youth could minimize. For this study (n=120) samples were collected from food contact surfaces i.e. work tops, equipments, tools, utensils and hands of food handlers working in cafeteria of various universities of Lahore. Each sample was analyzed for microbiological tests i.e. Total Aerobic Colony Count (ACC), Total Coliform Count, Staphylococcal count and Salmonella. The microbiological quality of various food contact surfaces of university cafeterias preparing ready to eat food was determined. Results highlighted poor hygienic status of FCSs in Universities. Among 120 samples 95.28% have been poor hygiene status according guidelines. Work tops and food handler’s hands had high total plate count value. Utensils was having low bacterial load than other two surfaces. Total Coliform count on utensils was lower than other two surfaces that are work tops and food handler’s hands.
Summary
40
Staphylococcus count on hands was abundant as compare to other two surfaces. In all the three microbiological tests Total Coliform Count was lower as compared to Total Plate Count and Total Staphylococcus Count. In sum the hygiene status of all the surfaces was highly unacceptable indicating that these surfaces are harboring many pathogenic bacteria that could be transferred to food that will come in contact in result of cross-contamination as food contact surfaces and one of three basic routes of cross contamination of food. It indicates that FCSs directly involve in food poisoning and food-borne outbreaks. Guidelines should be provided and implemented to reduce the risk of food contamination. Awareness and proper training and good hygiene practices are highly recommended in light of given results. Availability: Items available for loan: UVAS Library [Call number: 2728-T] (1).
57.
Development And Evaluation Of Vaccines Prepared From Staphylococcus Aureus Isolates Of Camel Mastitis
by Amjad Islam Aqib (2013-VA-947) | Dr. Muhammad Ijaz | Dr. Riaz Hussain | Prof. Dr. Aneela Zameer Durrani | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Development And Evaluation Of Vaccines Prepared From Staphylococcus Aureus Isolates Of Camel Mastitis Availability: Items available for loan: UVAS Library [Call number: 2750-T] (1).
58.
Isolation And Characterization Of Phytase Producing Fungi For Poultry Feed
by Ali Ahmad (2002-VA-121) | Prof. Dr. Aftab Ahmad Anjum | Prof. Dr. Masood Rabbani | Prof. Dr. Kamran Ashraf.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Isolation And Characterization Of Phytase Producing Fungi For Poultry Feed Availability: Items available for loan: UVAS Library [Call number: 2770-T] (1).
59.
Identification Of Genetic Marker In Gh, Igf-1 And Bmp15 Candidate Genes Associated With Growth Trait In Beetal Goat
by Mehwish Shareef (2009-VA-562) | Dr. Atia Basheer | Dr. Imran Zahoor | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Goats are pivotal indigenous assets of Pakistan and have an ample share in the annual production of milk and meat besides providing enough income to rural families. Beetal are famous for the meat and milk production and found throughout the Punjab province. But information regarding their genetic potential for meat production is still insufficientand need comprehensive genetic and genomic research. There is significant need to explore the candidate genes for growth traits in goat.GH, IGF-1 and BMP15 genes have been reported as candidate genes because they have a role in growth and skeletal development. It was hypothesized that polymorphism may be present in GH gene, IGF-1, and BMP15 genes which might also be associated with body weight and body measurements in Beetal goat. In the present study, 5 exons of growth hormone (GH) gene, exon 4 and partial region of intron 4 of IGF-1 gene and exon2 of BMP15 gene were amplified from goat genomic DNA.To assess this assumption, a total of 60 adult healthy animals of Beetal goat were selected on the basis of maximum variation in their body weight and body measurement (Body Length, height and heart girth). Animals were divided into two body weight categories, i.e. heavy and low.After blood collection (5ml), genomic DNA was extracted from the whole blood and stored at -20ºC for further use. DNA quantification was performed through agarose gel electrophoresis with the help of standard. PCR conditions were optimized. The genotyping was done through RFLP. General Linear Model (GLM) procedure of the Statistical Analysis System package (SAS Institute Inc, version 9.1) was used to test the effectof genotype on the body weight, body length, body height, and chest girth.Present study resulted in the identification ofsome SNP in GH, IGF-1 candidate genes in Beetalgoats. Genotyping results were monomorphic in GH, and BMP15 genes in animals of studiedbreed, while IGF-1 gene was polymorphic and had significant associationof genotypes with body weight and with body measurements. Genotype AA is significantly associated with low body weight having 38.18 Kg body weight and heart girth 33.65 cm, (P<0.05). AA genotype is also significantly associated with body leangth (P<0.05). The goats having BB genotype is significantly associated with high body weight having body weight 47.13 Kg (P<0.05). Genotype AB is significantly associated with withers height (31.93 cm).The sequencing results of the IGF-1 gene showed conserved and showed no other mutation in studied exonUponsequencing,polymorphism was found in GH gene inhigher and lower body weight animals. One SNP was identified in exon 2. The Non-synonymous substitution mutation (A > G) of CGA to CAA at position 825 caused an amino acid change from threonine into Alanine. While one deletion mutation in intron 4was identifiedat position 1546 in this gene. Up-to our knowledge, this is the first study of growth trait genes in the Beetal goat breed of Pakistan and helpful to understand the genetics of growth genes in Beetal goat breed. It may lead us to the identification of useful molecular markers for future use in the selection of animals with better growth traits.
CONCLUSION
By evaluating the genotyping and sequencingresults of all the target regions of growth hormone GH, IGF-1 and BMP15 genes, it is concluded that GHgene and BMP15 gene in beetal goat are monomorphic. One substitution mutation in exon 2 G>Aat position 825 along with one deletionin intron 4at position 1546 can be used as genetic markers in selection programme of breeding. WhileIGF-1 is of prime importance because IGF-1 gene was polymorphic and had significant association with body weight and measurements.
RECOMMENDATION
Due to limited resources 60 animals were used to determine polymorphism in three growth genesand their association with growth traits. It is recommended for further studies complete gene could be genotyped and sequenced and more number of animals should be used to evaluate these traits in future.
Availability: Items available for loan: UVAS Library [Call number: 2788-T] (1).
60.
Clinico-Epidemiological Study Of Multiple Drug Resistant Staphylococcus Aureus From Bovine Mastitis
by Muhammad Abdul Rauf Malik (2015-VA-832) | Dr. Muhammad Ijaz | Dr. Syed Saleem Ahmad | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Bovine mastitis is one of the most significant problems of livestock causing huge economic losses in dairy industry of Pakistan. Among other bacterial etiology of bovine mastitis, the Staphylococcus aureus is overwhelming to control and is well-known to cause subclinical and contagious mastitis. Methicillin resistant Staphylococcus aureus is our prevailing field issue. In view of the economic importance of Staphylococcus aureus mastitis, the current project was designed to study the Clinico-epidemiology of multiple drug resistant Staphylococcus aureus in bovine mastitis.
A total number of 900 milk samples (n=450 cattle, n=450 buffalo) were collected from Faisalabad district of Punjab, The collected samples were processed in laboratory of Microbiology and Medicine, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore. Primarily, screening of subclinical mastitis was done by Surf Field Mastitis Test (SFMT). Milk samples were spread out primarily on blood agar to rule out possibility of loss of later growth. Colonies with round and golden color characteristic were put to sub-culturing on Mannitol salt agar as differential and selective medium. The morphological and microscopic clarification was done under microscope using Gram’s staining technique. Various biochemical tests including coagulase and catalase were applied.
Prevalence of subclinical mastitis was found 55% (495/900), however significant differences were found among different tehsils of district Faisalabad. The prevalence of subclinical mastitis in cattle from district Faisalabad was found 54% (243/450) that upon comparison between different cities presented significant difference. While buffaloes presented 56% of subclinical mastitis. Comparisons of subclinical mastitis among different tehsils were
Summary
86
found significant.
Among quarter based prevalence of subclinical mastitis and prevalence of blocked quarters the number of blocked quarters were found 5.58% (201/3600) with highest percentage of blocked quarter noted in case of front right followed by rear right , front left , and rear left with 6.89, 6.56, 4.67, and 4.22%, respectively. However, the quarter based prevalence was found 32% (1088/3399) from bovine. The association of bovine subclinical mastitis with different risk factors presented significant association with few exceptions. Age, parity and body health status of animal were found non- significant with prevalence of mastitis. The breed and open rearing system presented significant relation with chances of mastitis. Chi-square test was used to statistically correlate the risk factors and prevalence of subclinical mastitis. P-value less than 0.05 was considered as significant.
Molecular confirmation of S. aureus was done by using coag gene through PCR technique. The S. aureus which were isolated from milk samples were put to antibiotic (oxacillin) sensitivity test for estimation of prevalence of methicillin resistant S. aureus from the bovine mastitis. Molecular identification of mec-A gene in staphylococcus aureus was done through PCR. The PCR confirmed methicillin resistant S. aureus isolates from cattle (n=20) and buffalo (n=20) were tested for their in-vitro drug response. However, Ciprofloxcin, Moxifloxacine, Linezolid, and Trimethoprim + Sulphamethoxazole were found 100% effective against multiple drug resistant Staphylococcus aureus and Levofloxacin showed 90% efficacy in bovine. While Oxytetracycline, Tylosin, Gentamycin, Amikacin, Vancomycin, and Fusidic acid were also found sensitive moderately except cefoxitin which was responsible for 100% resistane in bovine. Gentamycin was found to much more effective in buffaloes rather than cattle.
Summary
87
The study provided current status of Staphylococcus aureus infection with higher percentage in bovine mastitis. The prevalence of mecA gene revealed variation in methicillin resistant Staphylococcus aureus. In-vitro drug trial provided effective treatment possibilities against multiple drug resistance Staphylococcus aureus. Availability: Items available for loan: UVAS Library [Call number: 2815-T] (1).
61.
Development Of A Cost-Effective Serodiagnostic Assay For Peste Des Petits Ruminants (PPR)
by Tahira Hanif (2015-VA-1060) | Dr.Jawaria Ali Khan | Dr.Aamer Bin Zahur | Dr. Muhammad Avais | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Peste des petits ruminant (PPR) is a highly contagious newly developing disease of Small ruminants (sheep and goats). Currently the poor small ruminant’s farmers in Pakistan are facing huge economic losses due to PPR virus. In Pakistan PPR causes economic losses of Rs. 20.5 billion annually. The objectives of present study were to develop a cost effective sero-diagnostic assay for PPR (active haemagglutination inhibition and passive haemagglutination inhibition) and determination of comparative efficacy of active and passive haemagglutination inhibition assay (HI and PHA respectively) for detection of PPR virus infection.
In the present study, n= 300 sera samples were collected from sheep and goats during the (15 Februry 2016 to 2 January 2017).The serum samples were collected from kotli AJK :20(8 goats and 12 sheep),from gilgit:30 serum (20 goats and 10 sheep),from mansehra:22 serum (13 goats and 9 sheep),from mithi:112 (60 goats and 52 sheep) and 116 serum samples (88goats and 28 sheep) from Dhera ghazi khan.None of the animal was known to have been vaccinated against PPR previously or at the time of sampling.
These samples were collected from animals showed symptoms of PPR suggestive of PPR disease as well as from healthy animals. The sera were transferred into sterile tubes and were preserved on ice packs while shifting to the laboratory. PPR virus isolate was originally isolated from an outbreak in Taxila village, district Rawalpindi, the isolate was attenuated serially onto the Vero cell lines up to 20 passages. After, which antigen was titrated using a micotiter haemagglutination (HA) test with chicken RBCs and stored at -70◦c until use as a PPR antigen in a HI test.
In this study Active haemagglutination inhibition (HI) and passive haemagglutination inhibition were developed. The Haemagglutination Assay was standardized by different factors i.e. diluents, Temperature of incubation, Time of Incubation and concentration of Chicken R.B.C̓s. An additional test passive haemagglutination inhibition was performed to check the comparative efficacy of Active and Passive haemagglutination inhibition. In passive haemagglutination inhibition tanned sensitized cells remains effective due to their long effective life when stored at 4̊c and its makes an ideal test for diagnosis of PPR. Newly developed assays were compared against cELISA for PPR using kappa statistics and diagnostic sensitivity and specificity were determined.
The results of both assays were compared with results of competitive enzyme linked immunosorbent assay. In this study cELISA was considered as golden standard. The relative sensitivity and specificity of Active haemagglutination inhibition is 94.9% and 97.9% respectively. (Kappa 0.9264). However the sensitivity and specificity of Passive haemagglutination inhibition is 91.1% and 95.0% respectively.(kappa 0.8595). This study describes the serological detection of PPR virus by Active haemagglutination and passive haemagglutination inhibition (HI and PHA respectively). It was also concluded the comparative efficacy of (PHA and HI) that Active haemagglutination inhibition is more reliable technique than passive haemagglutination inhibition assay for the diagnosis of PPR disease in small ruminants (sheep and goats). Availability: Items available for loan: UVAS Library [Call number: 2816-T] (1).
62.
Physico-Chemical Factors Affecting In Vitro Stability And Activity Of Phytase From Indigenous Isolate Of Asperillus Therreus
by Safina kouser (2011-VA-422) | Dr. Aftab ahmad anjum | Dr.jawad nazir | Dr.Muhammad Yasir zahoor.
Material type: Publisher: 2017Dissertation note: Phytase is commercially important enzyme. Phytate in food and feed makes it less nutritive as well as acts as anti-nutritional agent. Phytate make complexes with important mineral ions and proteins. Monogastric animals and human are not able to degrade the phytate from plant based food because they lack phytase. This leads to phosphorous deficiency. Addition of phytase into food and feed degrades the phytate. It makes, phosphorous and mineral ions become available for growth and development. There is need to evaluate these factors in vitro which in real affect the stability and activity of enzyme under feed production process and digestive system of monogastric animals.
Indigenous Aspergillus terreus isolate produce stable phytase to be used in poultry feed.Indigenous strains of Aspergillus terreus were identified by macroscopic and microscopic characteristics. These isolates were screened on Phytate Screening Medium (PSM) for phytase production. Phytase producing A. terreus was than analyzed for toxin production through TLC (Thin layer chromatography). Non toxigenic phytase producing A. terreus isolates were inoculated in phytate broth for phytase production through submerged fermentation (SmF) under optimum conditions (28°C for 8-10 days). After centrifugation and filtration supernatants were used as crude enzyme. Phytase enzyme was qualitatively analyzed through phytase assay. Phytases activity units observed for isolate PAST-16 was highest (271.49±8.14 FTU/mL) and lowest (79.00±8.05FTU/mL) of PAST-05. A. terreus phytase (PAST-16) was subjected to temperature, pH and metal ions treatment.
Thermostability of phytases was recorted at 35°C, 55°C, 75 °C and 90°C for 15, 30, 45, and 60 minutes treatments. Enzyme from A. terreus (PAST-16) was observed as thermostable at
Summary
74
35°C, 55°C, 75 °C but not much stable at 95°C. Phytases showed 87.23±6.59, 198.34±4.47, 188.59±8.77 and 259.25±0.84 FTU/mL decreased in activity after 60 minutes of treatment at 35°C, 55°C, 75 °C and 95°C temperatures, respectively.
pH stability of phytases was found at pH of 2, 4, 6 and 8 for 15, 30, 45, and 60 minutes treatments. Enzyme from A. terreus (PAST-16) was observed as pH stable at 4, 6 and 8 but not much stable at 2 pH. Phytases showed 206.14±6.37, 169.59±6.37, 110.13±6.75 and 171.54±3.04 FTU/mL decreased in activity after 60 minutes of pH treatments at 2, 4, 6 and 8, respectively.
Metal ions effect on phytase activity was found with Ba2+, Ca2+, Cu2+, Fe3+, K+, Mg2+, Mn2+ and Na+ at the concentration of 1, 5 and 10mM. Enzyme from A. terreus (PAST-16) was observed as shows activity more with K+ less with Na+. Phytases showed 45.32±28.54 and 219.30±11.04 FTU/mL decreased in activity after 1mM conc. of K+ and 10mM conc of Na+, respectively.
Conclusion:
A.terreus isolate (PAST-16) produce stable phytase enzyme used in feed of poultry. In this way it tolerates condition under which feed process at commercial level and under digestive system monogastric animals. Availability: Items available for loan: UVAS Library [Call number: 2825-T] (1).
63.
Evaluation Of Antimicrobial Activity Of Essential Oil And Extracts Of Nigella Sativa Against Antibiotic Resistant Salmonella Enterica Isolates Of Human And Poultry Origin
by Sadia ashraf(2011-VA-402) | Prof. Dr. Aftab Ahmad Anjum | Dr. Ali Ahmad Sheikh | Dr.Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: The research was designed to evaluate the efficacy of methanolic and aqueous extracts of Nigella sativa, Black seed oil and thymoquinone against antibiotic resistant molecular characterized Salmonella enterica isolates of human and poultry origin (n=5 each). The compounds that have shown the antibacterial activity was also checked for their cytotoxicity by MTT assay.
Salmonella is causative agent of invasive diseases in poultry and humans, results in high mortality. Salmonellosis is a disease caused by Salmonella enterica with serious health issues related to food borne illness and most of world’s population is suffering from it. Mostly infections are treated by antibiotics but now a day’s resistance developed by Salmonella enterica. So it is need of time to develop some alternate ways to combat the problem caused by resistant bacterial pathogens. Use of essential oils and extracts of seeds are good weapons against resistant bacteria.
Salmonella enterica isolates of human and poultry origin (n=5 each) were taken from Department of microbiology UVAS Lahore and identified by colony morphology, microscopic characters, biochemical testing (Indole production test, Methyl red test, Voges Proskaeur test, Citrate utilization test and Urea utilization test) and polymerase chain reaction (PCR). For PCR product 1.5% agarose gel was run by gel electrophoresis. The biochemically identified and molecular characterized S. enterica isolates were screened for antibiotic susceptibility by Kirby Bauer disc diffusion method against amoxicillin, ampicillin, cefixime, ceftriaxone, ciprofloxacin, gentamicin, nalidixic acid, co-trimoxazole, ofloxacin and tetracycline and resistant pattern was 100% against ampicillin and Nalidixic acid and isolates shown 60% resistant against co-trimoxazole, amoxicillin and tetracycline, 80% and 40% resistant found against ofloxacin and ciprofloxacin while all isolates sensitive to cefixime and ceftriaxone. Aqueous and methanol were
CHAPTER 6
SUMMARY
used as solvents for extraction from Nigella Sativa. Seeds were dried, mixed, centrifuged, filtered and filtrate evaporated to obtained extracts. Percentage yield of methanolic extract was more than aqueous extract. Commercially available black seed oil, thymoquinone, water and methanol extracts of black seed would be evaluated for antibacterial activity by well diffusion method. Zones were measured in millimeters. All compounds gave the zones of inhibition except aqueous extract against Salmonella enterica isolates. The minimum inhibitory concentration (MIC) was determined by micro broth dilution method and then methanolic extract, black seed oil and thymoquinone used in MTT assay to evaluate their cytotoxicity. Cell survival percentage was calculated by a formula.
Data were analyzed by one way analysis of variance (ANOVA) followed by Duncan’s multiple range tests (DMRT) using statistical package for social sciences (SPSS version 20.0). Antimicrobial activity of essential oils, thymoquinone, water and methanol extract would be compared by graph pad prism 5.0 statistical software. Availability: Items available for loan: UVAS Library [Call number: 2827-T] (1).
64.
Production Of Polyhydroxybutyrate By Submerged Fermentation Using Agricultural By-Products
by Zainab Bibi (2015-VA-802) | Dr. Muhammad Tayyab | Dr. Shagufta Saeed | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Increasing non-degradable waste on planet is the major environmental concern these days. Hence, there is an absolute need of “eco-friendly” plastics. Polyhydroxybutyrate (PHB) is the most popular biodegradable as well as eco-friendly polymer. However, high production cost of PHB is still a major problem in the commercialization of biodegradable plastics. Process economics revealed that the use of cheap and renewable carbon substrates such as agro-industrial wastes can account for 40-50% reduction in overall production cost.
In present study wheat bran, gram bran, rice bran, wheat straw and sesame oil cake were used to check PHB production using Azotobacter vinelandii NRRL-146641. For this purpose, 0.5ml inoculum was added in fermentation media and kept for incubation at 24-48 hrs. After incubation, both physical and chemical parameters such as (substrate water ratio, incubation time, inoculum volume, pH, agitation rate and nitrogen sources) were optimized. Optimized culture medium was centrifuged and obtained sediment was then used for analysis.
It was found that Azotobacter vinelandii in Rice bran contained medium gives maximum yield of PHB (248mg/100mL) at 8% substrate water ratio after 96 hours of incubation period (292mg/100mL), at 1.5 mL of volume of inoculum (304 mg/100mL), at pH 6.0 (316 mg/100mL), at 160 rpm agitation rate (416mg/100ml) at 0.3 % of yeast extract (446 mg/100mL) and 0.25% (436mg/100mL) of peptone. Obtained data was then analyzed by means of ONE-WAY ANOVA and through LSD test. Availability: Items available for loan: UVAS Library [Call number: 2855-T] (1).
65.
Acaricide Resistance Of Tick Population Infesting Buffaloes In District Narowal
by Muhammad Mubashir Abdullah (2015-VA-1104) | Prof. Dr. Kamran Ashraf | Dr. Muhammad Latif | Prof. Dr. Aftab Ahmad Anjum.
Material type: Publisher: 2017Dissertation note:
Tick imperviousness to acaricides is an expanding issue in Pakistan and represents a genuine financial danger to the domesticated animals and veterinary pharmaceutical enterprises. New acaricides are to a great degree costly to grow so the present acaricides ought to be viewed as a constantly decreasing asset, which ought to be ensured by all methods conceivable. The principle goal of the review was to distinguish the stages of tick imperviousness to acaricides at close business and collective ranges in District Narowal, Pakistan. Likewise to contrast the in vivo techniques and with explore acaricide administration procedures which may build the life expectancy by utilized acaricides.
To meet these points a field survey (February 2016 to March 2017) was carried out at 3 tehsils (Tehsil Narowal, Shakargharand Zafarwal cities of Pakistan to monitor levels of field tick resistance to acaricides. The larvae were originally obtained from engorged female A.hebraeum, Hyalloma, Boophilus, Dermacentor, Ixodes, R. appendiculatusand R. evertsievertsi. The larvaewere tested against different concentrations of trichlorofon, ivermectin and cypermethrin using the Shaw Larval Immersion Test (SLIT). Mortality dose data were subjected to probit analysis using a BMDP statistical package. Factors of resistance (FOR) were calculated by comparing the larval response of ticks from the field.
On the communal farms high levels of tick resistance were detected to cypermethrin as well as partial resistance to ivermectin whilst no resistance was detected against trichlorofon. On the commercial farms, however, ticks were equally resistant to trichlorofon, cypermethrin and ivermectin. The populations of Hyalloma, Boophilus, Dermacentor, Ixodes, on these farms had developed higher levels of resistance to the testacaricides than the equivalent R. evertsievertsi, R. appendiculatus and A.hebraeumpopulations. Higher levels of tick resistance to trichlorofonwas observed on3 tehsils (Tehsil Narowal, Shakarghar and Zafarwal)than on communal farms, however, there was no significant differences in tick resistance to ivermectin and cypermethrin at both the commercial and communal farms. It was surmised that inappropriate use of acaricides might have resulted in higher tick resistance to the currently available acaricides on the commercial as well as the communal farms. Correct acaricide usage may solve this problem to a limited extent.
Comparative in vivo tests were also carried out on the larvae and adults of Hyalloma, Boophilus, Dermacentor, Ixodes, to determine the susceptibility of this tick to different concentrationsof the currently used acaricides, (amitraz, ivermectin and cypermethrin) at three commercial dairy farms, (“Brycedale”, “Sunny Grove” and “Welgevind”) in the areas of District Narowal, Pakistan. Resistance of field strains of Hyalloma, Boophilus, Dermacentor, Ixodes, Dermacentor,were determined using the Adult Immersion Test (AIT) as the latter test took into account factors such as oviposition assessment and reproductive ability. At “Brycedale”, resistance to trichlorofon and ivermectin was detected with the AIT method. Emerging resistance to trichlorofon and resistance to ivermectin were also detected . At “Sunny Grove” resistance was detected to cypermethrin and at “Welgevind” resistance was detected to ivermectin with the SLIT whilst no resistance was detected using AIT. It would appear that the Hyalloma, Boophilus, Dermacentor, Ixodes, populations tested on these dairy farms were more resistant toivermectin than to trichlorofon or cypermethrin.
Nearly 50% of the dairy farms sampled showed resistance to ivermectin and the majority had susceptible Hyalloma, Boophilus, Dermacentor, Ixodes, populations to both amitraz and cypermethrin. In general there was a good correlation between the Cypermethrin and Trichlorofon whilst in many cases there was poor correlation between the Cypermethrin and Ivermectin.
From this study it would appear that the In vivo method was a reliable to detect resistance within seven days. In vitro method the ELT and the RET could possibly be used as screening methods to detect acaricide resistance on farms whilst the SLIT would remain the test of choice for National surveys. In addition the ELT is less costly and does not require sophisticated equipment for field testing if resistance development compared with other in vitro test methods. This method, however, still needs to be validated and standardized for use in Narowaland the rest of punjab where tick control is important.
Availability: No items available
66.
Determination Of Tartrazine In Different Food Items Available In Lahore Pakistan
by Anam Arshad (2015-VA-1055) | Dr Zubair Farooq | Dr. Sanaullah Iqbal | Prof. Dr. Aftab Ahmad Anjum.
Material type: Publisher: 2017Dissertation note: Synthetic dyes used in various food items like sweets, candies and beverages cause severe health hazards if they are not food grade and within the legally permitted limits. In Pakistan due to increased consumption of attractive colored food items, the use of dyes in every food product is also increasing. Already there is no study to appraise the nature and level of colorants. Therefore, this study focused on determination of synthetic dye (tartrazine) used in candies, sweets and beverages to determine its safer levels.
This research work was done in food science and nutrition lab of Food Science and Human Nutrition Department of UVAS, Lahore, Pakistan. Total 180 samples were collected from all 9 towns of Lahore plus Lahore Cantt. Samples included 30 candies purchased from local vendors and 30 candies from shops, 30 sweet samples (rasgulla) from local vendors and 30 sweet samples from sweet shops. Moreover, 30 slush samples locally available in streets and 30 slush samples from shops of posh areas in Lahore. The results were compared with the previous held researches in other countries.
The food samples were divided into two categories local shops and local vendors. Total six local shops and six vendors of each town of lahore were selected for the collection of samples in triplicate pattern. All the samples were evaluated for spectrophotometric determination of tartrazine by using Beer’s law. Abosrbtion peeks were checked at a wavelength of 421.6 and the mean values of concentration of tartrazine in slush ranged from 0.269 to 0.275 mg/g obtained from local vendors and shops respectively.Moreover, the mean values of tartrazine ranged from 0.258 to 0.309 mg/g for vendor sweet and shop sweet samples respectively and mean value for candies ranged from 0.200 -0.704mg/g. Data was analyzed through the Microsoft Excel 2010 and SPSS 22.0. Mean values with standard deviation in percentages of results were analyzed by descriptive analyses. To examine the relationship among the variables of candies, sweets and slush samples chi-square test was used. Further to compare tartrazine levels in the local shops and foodstuff obtained from the common vendors of Lahore, independent “t” test was used. 2 way-ANOVA was applied to check the differences of all samples in 10 towns of Lahore.
According to my investigations the quality of foodstuff collected from local shops and from local vendors is almost same and both contain high amounts of tartrazine.I suggest consumers, they should prefer to buy branded foodstuff from superstores as compared to local shops and local vendors because the keeping quality and handling practices are good in superstores than local shops.
Availability: Items available for loan: UVAS Library [Call number: 2886-T] (1).
67.
Mutation Detection Of Cacnb4 Gene Involved In Childhood Absence Epilepsy And Its Comparative Genomics In Mice
by Ayesha Amin (2015-VA-1048) | Dr. Muhammad Wasim | Dr. Sehrish Firyal | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Childhood absence epilepsy (CAE) is due to multiple factors affecting almost 9-12 years children. Depolarization of ion channel activates the channel. CACNB4 gene is affected by epileptic seizures. Disturbance in ion channel can affects different genes as CACNA1G, CACNA1H, CACNA1A, SCN1B, SCN1A,SCN2A and GABA receptor genes. CACNB4 gene has a major role in influencing epilepsy in human.
In present study,it is directed to analyze the mutations in epilepsy present in coding region of CACNB4 gene.
Collection of blood samples were from Children Hospital, Lahore, Punjab Pakistan from CAE patients of epilepsy. By using standard DNA extraction method, DNA was extracted from samples. Primers were designed for the amplification of exon 3 and 13 of CACNB4 gene.
Results were examined after sequencing the samples. BioEdit software was used to study the samples thoroughly. NCBI BLAST was used to align the sequences.
It is investigated that the sequences of CAE patients of epilepsy of CACNB4 gene has mutation at position position 258023bp which changes A>G. In protein sequence, the mutation is at position 413 which changes L (Leucine) to L (Leucine). This mutation has no effect because this is a synonymous mutationwhere the codon CUG is changes to CUA, both codes for same amino acid that is leucine, so no effect at all by this change in exon 13.
Three mutations are present in the intronic region of exon 13 first, second and third at positions 258184bp A deleted, 258289bp and 258191bp of CACNB4 gene respectively. These all mutations are present in intronic region so has no effect in phenotypes of individual.
In conclusion, maximum numbers of samples were needed to observe the effect of mutations and factors that causes epilepsy. This study will now help the researchers to investigate genetic therapies, strategies of genetic counseling and parental diagnosis for the population of Pakistan.
Availability: Items available for loan: UVAS Library [Call number: 2924-T] (1).
68.
Spatial Ecology And Distribution Of Soil Borne Burkholderia Mallei In Punjab, Pakistan
by Muhammad Asad Ali (2002-VA-73) | Prof. Dr. Khushi Muhammad | Prof. Dr. Aftab Ahmad Anjum | Prof. Dr. Mansur-Ud-Din Ahmad.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Burkholderia mallei is a causative agent of glanders, the disease of equines. The disease is characterized by pulmonary, nasal and cutaneous forms. B. mallei is excreted through nasal discharge, lacerated skin/wounds and expiration. Diseased animals shed bacteria through the discharges contaminating soil, water, fodder and other susceptible animals in its vicinity. The present study was designed to map and investigate the association of different physical factors and soil chemistry analytes with persistence of B. mallei genome in soil of 10% percent villages (n=456) from eight selected districts of Punjab province, Pakistan.
Eleven (0.48%) out of 2, 280 soil samples were positive for B. mallei genome in varied locations of Punjab. Higher prevalence (2.37%) for genome was detected in Sheikhupura district followed by Chakwal district (2.10%). None of the samples from Gujranwala, Sahiwal, DG Khan, Attock, Faisalabad and Sargodha districts were found positive for B. mallei genome. The genome of B. mallei was distributed in 25% study districts of Punjab, Pakistan. In Chakwal district, the genome of B. mallei was strongly associated with moisture (p=0.008) in all positive samples ranging from 0.80 to 39.20%, Phosphorous (p=0.050) ranging from 1.74 to 21.75 mg/Kg. While, this association in Sheikhupura district soil samples was with Sodium (p=0.018) and moisture (0.026) ranging from 1.90 to 133.59 mg/Kg and 0.80 to 39.20%, respectively. The odds of detecting DNA of B. mallei were recorded higher (1.4, 6.8, 5.0, 2.8 and 10.6 ) when soil sample sites were < 500 meters away from vehicular traffic roads, < one kilometer from animal markets, < 100 meters from canal, animal density < 1,000 animals and human population < 300 houses/village. While the odds of detecting DNA of B. mallei were 0.1, 0.3, 0.4, 0.2 and 0.5 when soil sample sites were > 500 meters from vehicular traffic roads, > one kilometer from animal markets, > 100 meters from canal, animal density > 1000 animals and human population > 300 houses/village, respectively. Soil-borne B. mallei DNA is more likely to be detected in areas closer to roads with vehicular traffic along the interstate routes in Punjab and soil containing low level of moisture.
It was concluded that soil of two districts out of eight selected was positive for B. mallei genome in Punjab province. Odds of less distance from main road to animal farm and high animal density at farm were positively associated with B. mallei DNA persistence in soil. Moisture, sodium and phosphorus were positively associated with persistence of B. mallei DNA in soil.
Availability: Items available for loan: UVAS Library [Call number: 2900-T] (1).
69.
Prevalence, Molecular Diagnosis And Chemotherapy Of Degnala Disease In Large Ruminants Of Punjab.
by Mudassar Nazar (2005-VA-92) | Prof. Dr. Muhammad Sarwar Khan | Dr. Muhammad Ijaz | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: In Pakistan, Livestock is considered as a social security for poor villager as it can be a source of cash at the time of need. Degnala disease reduces the production of these animals directly. Along with other side issues related to Degnala disease, this study was done to diagnose the actual cause of Degnala disease by applying different latest scientific techniques. Prevalence along with risk factors was calculated in the rice growing areas of Punjab, Pakistan. Fungal isolation (n=40) was performed from the rice straw feedings of the Degnala disease affected animals through the technique of spot culture on SDA. Then these fungal isolates were identified through comparing their microscopic and macroscopic characters. Then toxigenic potential was checked for all these isolates through the application of TLC and HPLC. After that, from those isolates which were positive for mycotoxin production potential, most cytotoxic isolate was checked with the application of MTT assay. Then the most cytotoxic isolate was inoculated on non-contaminated rice straw and fed to the experimental animals to see a similarity of natural cases of Degnala disease. Finally treatment was conducted to see a proper combination of various drugs against this disease.
Toxigenic potential of different candidate fungi, isolated from rice straw feeding of Degnala disease affected bovines was analysed along with Species, age, gender and season wise prevalence. Out of 1536, 104 (6.77%) cases showed positive signs for this disease with a significant association (p<0.05) between rice straw feeding in buffaloes, winter season and bovines having an age of more than one year. Complete Blood Count showed marked increase in erythrocyte sedimentation rate and all white blood cells numbers, except lymphocytes in positive cases. There was a significant increase (p<0.05) in Alanine amino transferase, Aspartate amino transferase and Alkaline phosphatase noticed in Liver Function Test. At the same time, increased value of Creatinine was noticed in Renal Function Test. For isolation and screening of toxigenic fungi, rice straw samples (n=40) being fed to the positive cases were processed further, out of which there were 85 fungal isolates mainly of Aspergillus (57), Penicillium (10), Fusarium (04), Zygomycetes (03), Curvularia (01) and unidentified (10). All isolated fungi were subjected for mycotoxin production and only 11 showed mycotoxin producing capability (including Aspergillus, Penicillium and Fusarium isolates) analysed by Thin Layer Chromatography and quantified through High Performance Liquid Chromatography. It is concluded that all the fungi, contaminating rice straw feeding of Degnala affected animals are not toxigenic. This work will help in establishing major mycotoxin producing fungi leading to the probable cause of Degnala disease in bovine.
With the help of MTT assay on vero cell line, most cytotoxic fungus was identified. After an incubation with vero cells, OD values of all the candidate fungi were compared through one way ANOVA. Results of this analysis showed that Fusarium was at the highest ranking and then was the A. flavus with a significant value of 0.006 and 0.039.
Finally it was concluded through these systematic steps of converging the diagnosis that, out of all the 85 suspected fungi, Fusarium (isolate number S 8.1) was the most cytotoxic isolate obtained from the rice straw feedings of Degnala affected animals in our study.
For molecular diagnosis of the most cytotoxic isolate of Fusarium, PCR was conducted and the results showed that ultimately the final PCR product was successfully amplified against the mentioned primer of ITS conserved region for Fusarium genera and the DNA product was with a length of 570 base pairs.
Experimental feeding trials were conducted by inoculating Fusarium (the most cytotoxic isolate) and A. flavus (second most cytotoxic one after Fusarium) separately and in combination compared with the negative control group, all groups were of eight animals each. It was concluded that alone Fusarium was able to produce Degnala disease, while its combination with A. flavus was more lethal.
Ultimately the treatment trials proceeded with penta-sulphate, oxytetracycline and antiseptic topical application as therapeutic treatment were shown to be very effective against Degnala cases. While in all the affected animals feeding of affected rice straw was ceased. Only withdrawal of affected rice straw from the feedings of Degnala affected animals was not effective unless proper treatment as mentioned here was not conducted.
analysed The expected results of the study shall be helpful to make exact diagnosis and treatment of infected buffaloes and cattle that is further helpful for timely prophylaxis and control of the Degnala disease in the rice growing areas of Pakistan and South Asia.
Availability: Items available for loan: UVAS Library [Call number: 2960-T] (1).
70.
Evaluation Of White Sesame Seed Oil As A Functional Food Ingredient And Its Role To Mitigate Hyperglycemia
by Farhan Aslam (2011-VA-606) | Dr. Sanaullah Iqbal | Dr. Muhammad Nasir | Prof. Dr. Aftab Ahmad Anjum.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: White sesame seed oil (WSSO) contains appreciable amount of various bioactive components including tocopherols, polyphenols, phytosterols and lignans (sesamin & sesamolin) known to have positive impact against certain diseases. Characterization of white sesame seed oil (PB Til-90) showed the presence of bioactive components make it suitable for human consumption. The comparison of WSSO based functional cookies and vegetable fat (VF) based cookies showed that energy and fat% were significantly higher (P < 0.05) in WSSO than VF cookies. At 60th day, mean moisture, peroxide value, and acidity were higher (P < 0.05) in VF cookies. Over time, protein and fiber% decreased significantly (p < 0.05) in both cookies but remained higher (P < 0.05) in WSSO at 60 days. By the end of the 60 days of storage time, moisture content in SO cookies increased approximately 34% (p < 0.05), while other components decreased significantly (p < 0.05) over time; (protein: -0.2%, fat: -3%, fiber: -5.5%, and ash: -7.9%). At 60 days there were significant (p < 0.05) differences between groups. Moisture was significantly higher in VF verses SO, whereas all other components were significantly (p < 0.05) lower in VF group compared to SO group; (protein: -7.6%, fat: -9%, fiber: -5% and ash: -11 %).
Over time, from baseline to 60 days, peroxide value increased approximately 252% in SO cookies. Additionally, in SO, acidity, nitrogen free extract, and thiobarbituric acid values increased (35%, 3%, 54% respectively), while bioactive components, sesamin and sesamol, decreased significantly (p <0.05) over time (i.e., -0.22% and -1.2% respectively). A similar trend was observed in VF cookies. Over the period from baseline to 60 days, the mean rating on each attribute decreased significantly (p < 0.05) for each cookie type. For SO cookies, colour decreased by about -5.5%, flavour -8%, taste -16%, texture -11.6%, crispness
SUMMARY
179
-8% and overall acceptability by -14%. A similar trend was observed in VF cookies. In VF cookies, the mean rating for colour decreased -9%, flavour decreased by -11%, taste decreased by -11%, texture decreased by -12%, crispness decreased by -7% and overall acceptability decreased by -5.5%. By day 60, there were significant (p < 0.05) differences in the sensory rating between groups.
For efficacy study on rats, sixty-three male Sprague-Dawley rats were randomized into standard diet groups (normal control, NCON, n=21) and (diabetic control, DCON, n=21) and a diabetic sesame oil (DSO) (n=21) group which were fed a diet containing 12% WSSO. Blood samples were analyzed at 0, 30 and 60 days. Differences between groups and across days were assessed with two-way repeated measures ANOVA. At baseline, GLU and INS were similar in both diabetic groups (mean 248.4 + 2.8 mg/dl) and (mean 23.4 ± 0.4 μU/mL) respectively. At 60 days, GLU was significantly (p < 0.05) higher in DCON (298.0 ± 2.3 mg/dl) as compared to DSO (202.1 ± 1.0 mg/dl). Activities hepatic antioxidant enzymes increased significantly (p < 0.05) in each variable across time from baseline to 60 days; SOD: (9.7 ± 0.1 to 15.5 ± 0.6 IU/mg), CAT: (6.6 ± 0.1 to 12.5 ± 0.8 IU/mg), GPx (11.1 ± 0.3 to 35.9 ± 3.2 IU/mg), APx (48.7 ± 1.6 to 76.1 ± 1.9 IU/mg) in the DSO group as compared to the DCON and NCON groups. In the DSO group, CK decreased significantly (p < 0.05) from baseline (291.1 ± 0.9 U/L) to 60 days (245.5 ± 7.2 U/L) from both the control groups, while CK-Mb decreased significantly (p < 0.05) from baseline (550.5 ± 3.9 U/L) to 60 days (510.8 ± 6.8 U/L) from NCON group but was not significantly different from DCON group. Among liver function tests, ALP increased over time in both diabetic groups (i.e., in DSO group from baseline to 60 days it raised from 246.7 ± 3.3 U/L to 277.7 ± 2.8 U/L) and at 60 days was significantly higher (p < 0.05) than NCON in both groups but were not significantly different from each other. In contrast, ALT from baseline (81.5 ± 3.7 U/L) to 60 days (67.4 ± 2.7 U/L) and AST from baseline (148.7 ± 3.5 U/L) to 60 days (118.3 ± 1.2 U/L) significantly decreased
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(p < 0.05) in the DSO group as compared to DCON or NCON resulting in significantly lower values than both control groups by 60 days. At 60 days, urea in the DSO group decreased from baseline (38.5 ± 2.3 to 30.9 ± 1.1) such that it was significantly lower (p < 0.05) than both control groups. From baseline to 60 days, creatinine significantly increased (p < 0.05) in the two diabetic groups; in DSO group at baseline creatinine was (0.3 ± 0.0 mg/dl) and increased up to (0.4 ± 0.1) after 60 days whereas it remained fairly stable in the NCON group. At 60 days, creatinine was significantly higher in both the diabetic groups as compared to NCON. At 60th day; cholesterol, triglyceride, VLDL, and LDL was significantly lower (p < 0.05) and HDL significantly was significantly higher than DCON, and NCON. The results indicated that there were no significant differences between the DSO or DCON groups in electrolyte balance, minerals, and hematological values.
For efficacy study on humans, forty-six subjects with Type 2 diabetes were recruited and randomly divided into two equal groups (diabetic control, DCON) and diabetic sesame oil (DSO). At baseline, 30, 60, and 90 days, blood samples were drawn and analyzed. Two-way repeated measures ANOVA was used to evaluate the difference between groups and across time. In both groups GLU, INS, and HbA1c were not significantly different at baseline; (mean 187.07 + 5.63 mg/dl), (mean 12.12 ± 1.03 μU/mL), and (mean 7.55 + 0.37 %) respectively. At 90 days, GLU was significantly (p < 0.05) decreased in DSO (137.83 ± 3.16 mg/dl) when compared with DCON (218.13 ± 5.92 mg/dl) while insulin was significantly increased in DSO (23.13 ± 1.15 U/ml) as compared to DCON (7.93 ± 0.38 U/ml). At 90th day HbA1c was significantly lower (p < 0.05) in DSO as compared to DCON. TBARS was significantly lower (p < 0.05) in DSO (1.08 ± 0.05 [MDA] nmol/ml) as compared to DCON (2.26 ± 0.07 [MDA] nmol/ml). In DSO, activities of hepatic antioxidant enzymes (SOD, CAT, and GPx) increased while in DCON these activities decreased significantly (p < 0.05) across time period. Biomarkers of liver, cardiac and renal functions improved significantly in
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DSO as compared to DCON. At 90th day; cholesterol, triglyceride, VLDL, and LDL were significantly lower (p < 0.05) and HDL was significantly higher than DCON. There were no significant differences between the DSO or DCON groups in electrolyte balance, minerals, and hematological values.
Conclusion:
It was concluded that consumption of white sesame seed oil significantly improved blood glucose regulation, reduced oxidative stress, improved antioxidant activity and biomarkers hepatic, cardiac and liver enzymes in male sprague dawley rats and type 2 diabetic patients. Availability: Items available for loan: UVAS Library [Call number: 2950-T] (1).
