1.
Production And Evaluation Of Peste Des Petits Ruminants Virus Vaccine
by Muhammad Aness | Prof. Dr. Khushi Muhammad | Prof. Dr. Masood Rabbani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: PPR is an acute highly contagious viral disease of small ruminants which is endemic in Pakistan. Present study was aimed to evaluate the freeze dried PPRV vaccine with variable biological titer to induce protective immune response in beetal goats. The comparative immune response of animals to adjuvant and non-adjuvant vaccine and duration of immunity was also studied. Effect of boosting on the humoral immune response of animals as well as shelf life of various vaccines was also evaluated. Each of the vaccines was inoculated in a group of five animals. Serum samples were collected at specified time intervals and antibody levels were detected through cELISA as PI values and neutralization test as MNA titer.
The virus was propagated on the Vero cells. It was estimated that infecting 2 x 107cells with 104.00 TCID50 virus concentrations added to a T-175 cell culture flask at the time of subculture yielded maximum virus titer in the cell culture harvest following three freeze thaw cycles of the contents. The freeze dried vaccine with a biological titer of 105.00 TCID50 per dose provoked maximum antibody titer followed by the ones with a titer of 104.00 or 103.00TCID50 which provoked nearly equivalent protective immune response while the animals inoculated with a vaccine having 102.00 TCID50virus concentrations developed minimum antibody titer. The oil adjuvant PPRV vaccines elicited significantly higher antibody titer in comparison to gel based vaccines but however minimum antibody titers were detectable in response to freeze dried vaccines. Although protective antibody level (? 10 neutralizing antibody units) was detectable in the animals vaccinated with either oil based, gel based or freeze dried vaccine containing biological titer of 104.00 TCID50 but however the extent and duration of immunity was found to be most superior in response to oil based vaccines. It can be concluded that a single shot of either gel or oil based vaccine can provide protection in the vaccinated animals for a minimum of one year duration. Goats receiving a booster dose of the vaccines had a significantly higher antibody tier in comparison to the ones who received single dose of the vaccines. The freeze dried and wet vaccine kept at 4 °C did not show any significant drop in the biological activity of the virus even after 12 months of storage. Immunogenicity of the both adjuvant and non-adjuvant vaccines, as measured through the immune response in the vaccinated animals, also remained unaffected after 12 months of storage at 4 °C.
Availability: Items available for loan: UVAS Library [Call number: 1731,T] (1).
2.
Molecular And Serological Characterization Of Avian Influenza Viruses In Domestic And Wild Birds
by Mobeen Sarwar | Prof. Dr. Khushi Muhammad | Prof. Dr. Masood Rabbani.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Avian Influenza virus (AIV) has been recognized one of the most important pathogen in poultry industry. AIVs play an important role in the pandemic spread that could cause high morbidity and mortality in human beings. A total of 1500 tracheal and cloacael swabs were collected from the seven live bird poultry markets of Lahore Pakistan for surveillance of AIV. The samples were processed for virus isolation in chicken embryos. The isolates were processed for HA test and AIV Antigen Rapid Test Kit to differentiate Newcastle disease virus and Avian influenza virus. Only four samples were positive for Avian influenza H9N2 subtype and 17 were positive for Newcastle disease virus. Four HA virus suspension of AIV showed high titers of anti AIV H9N2-HI antibody titer in chicken as well as rabbit serum, raised using Ottoman Pharma AIV-H9N2 (Oil based) vaccine.
The isolates of AIV H9N2 were confirmed using laboratory optimized RT-PCR, mRT-PCR and LAMP tests. All isolates were sequenced and analyzed to develop phylogenetic relationship. Two isolates A/Chicken/Pakistan/Micro-1/2009 and A/ chicken/ Pakistan/ Micro-2/ 2009 showed 99.1% nucleotide homology with each other and 95- 99% homology with the other Pakistani isolates, 95.1% homology with A/Chicken/Iran/B102/2005. The nucleotide sequence of "A/Duck/Pakistan/Micro-3/2009" showed 98.8% homology with "A/chicken/Pakistan/micro-4/2009", 98-98.7% homology with other Pakistani Isolates and 95.8% homology with "A/Chicken/Iran/B102/2005". The nucleotide sequence of "A/Chicken/Pakistan/Micro-4/2009" showed 99.6% homology with "A/duck/Pakistan/micro-3/2009", 95-96% homology with other Pakistani isolates and 94.3% homology with "A/Chicken/Iran/TH lBM863/2007".
Three out of four isolates had PARSSRGL cleavage sites and one isolate A/chicken/Pakistan/micro-1/2009 had PAKSSRGL cleavage site. The four isolates of the study contained a 226-Leu and 228-Gly at receptor binding sites. Substitution of Glutamine (Q) into Leucine (L) at 226 receptor binding site in HA glycoprotein increased the binding specificity for Sialic acid ? 2, 6 Galactose linkage of human receptor. The HA gene of the live poultry market isolates had 7 predicted glycosylation sites at 29-32, 105-108, 141-144, 298-301, 305-308, 492-495 and 551-554, positions. The NA gene of the live poultry market isolates had 8 predicted glycosylation sites at 44-46, 61-63, 69-71, 146-148, 200-203, 234-237 and 402-403, positions. Predicted glycosylation sites affected the structure and stability of NA protein.
It is concluded that continuous epidemiological and virological surveillance of live bird poultry markets may help scientists to develop an effective control and preventive measures for AI viruses.
Availability: Items available for loan: UVAS Library [Call number: 1536,T] (1).
3.
Factors Affecting Biomass Producation Of Baby Hamster Kidney Cell Line In Roller Bottle Culture System For The Production of Foot and Mouth Diseas Virus
by Qaiser Akram | Prof. Dr. Khushi Muhammad | Prof. Dr. Masood Rabbani.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Foot and Mouth Disease (FMD) is endemic in Pakistan and killed virus vaccines against prevalent serotypes are used for the control of disease. Baby Hamster Kidney (BHK-21) cells as monolayer culture are routinely used for the propagation of FMD viruses. Various nutritional factors: amount of growth medium and serum concentration as well as physical conditions: seeding density, rolling speed, growth time, and incubation temperature for the propagation of BHK-21 cells on roller culture bottles were optimized. The roller culture bottles having surface area of 480 cm2 were used for the propagation of cells. Feeding of cells with 100 ml of growth medium per bottle and supplementation of 5% serum supported the growth of the cells in optimum way. Seeding of ten million cells per bottle resulted into the development of complete monolayer with maximum cell density within 48 hours. The cultured cells remained confluent up to 60 hours while a rapid decline in the number of harvested cells was recorded after 72 hours of incubation. Growth rate of the cells was slower at 33° C that increases at 35° C, reaching to its maximum at 37° C while cells could not tolerate the temperature of 39° C. The bottles kept at rolling speed of 3 rpm yielded maximum amount of cells while higher or lower rotation speed negatively affected the cell proliferation.
Antibody response of buffalo calves to different levels of FMD virus immunogen in trivalent vaccine was investigated. The vaccine containing 106.2 units of immunogen/TCID50 of each of the three serotypes of FMD virus induced log2(1.3± 0.4) units of anti-FMD "O" Complement Fixing Cumulative Geometric Mean antibody (FMD"O" CFT-CGM) titer, log2(1.4±0.3) units of anti-FMD"A" CFT-CGM titer and log2(2.0±0.7) units of anti-FMD"Asia-1" CFT-CGM titer. The vaccine containing 2x106.2 units of immunogen of each of the three serotypes of FMD virus induced log2(2.2±0.2) units of anti- FMD"O" CFT-CGM titer, log2(2.1±0.25) units of anti- FMD"A" CFT-CGM titer and log2(3.4±0.8) units of anti-FMD"Asia-1" CFT-CGM titer. The vaccine containing 3x106.2 units of TCID50 of each of the three serotypes of FMD virus induced log2 (5.3 ± 2.0) units of anti-FMD"O" CFT-CGM titer, log2 (4.6±1.9) units of anti-FMD"A" CFT-CGM titer and log2 (5.0±2.2) units of anti- FMD"Asia-1" CFT-CGM titer. The vaccine containing 4 x 106.2 units of TCID50 of each of the three serotypes of FMD virus induced log2(5.5±1.5) units of anti-FMD"O" CFT-CGM titer, log2(4.4±1.9) units of anti-FMD"A" CFT-CGM titer and log2(5.2±1.9) units of anti-FMD"Asia-1" CFT-CGM titer. Moreover, buffalo calves (n=3) which were primed and boosted with 60 days interval using vaccine containing 2x106.2units of immunogen of each serotype of FMD virus, showed log25.0 and log26.3 units of anti FMD"O"-CFT-GMT antibody titer, log24.6 and log2 6.0 units of anti FMD"A"-CFT GMT antibody titer, log25.6 and log26.0 units of anti FMD"Asia-1"-CFT GMT antibody titer, on 30 and 120 days post boosting.
Each serotype of the virus grew well on BHK-21 cell line. The virus showed poor TCID50 (log 103.2±0.2) in BHK-21 cells having Glasgow Minimal Essential Medium (GMEM) without Fetal Calf Serum (FCS). Addition of FCS in the medium at the rate of one percent increased log 107.1±0.2 units of virus TCID50. Incubation temperature of 350 C and 370 C supported the multiplication and maintenance of BHK-21 cells and yielded log106.6±0.1 and log107.0±0.2units of virus TCID50, respectively. Each serotype of FMD virus showed log106.29±0.07 units of virus TCID50 in the stationary monolayer of BHK-21 cells in Roux flask (75cm2), log107.66± 0.02 units of virus TCID50 in roller bottles (490 cm2) and log108.34 ± 0.07 units of virus TCID50 on 0.2 g of micro-carriers suspending in 200 ml of the growth medium in stirring bottle. The infectivity titer/TCID50 of each of the virus serotypes was significantly higher in roller bottles than that achieved in Roux flasks (p<0.05) and was significantly higher in stirring bottle containing micro-carriers suspending in the growth medium than that of harvested in roller bottle (p<0.05). It is concluded that the infectivity titer of the virus is directly proportional to number of BHK-21 cells in the culture system.
Availability: Items available for loan: UVAS Library [Call number: 1554,T] (1).
4.
Antimicrobial Effect Of Various Herbs On Sore Throat Causing Antibiotic Resistant Bacterial Isolates
by Abida Mushtaque (2010-VA-298) | Dr. Ali Ahmad Sheikh | Prof. Dr. Masood Rabbani | Dr. Muhammad Nasir .
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Sore throat or pharyngitis in childrenis mainly caused by group A (β-hemolytic) Streptococcus pyogenes. The bacterium transferred from infected person to susceptible one through coughing, sneezing and by direct contact. Irrational use of antibiotics to control sore throat infection results in development of antibiotics resistance. To overcome the problem of antibiotic resistance, various herbal extracts(Liquorice, ginger, cinnamon and red dates)are used which have known antimicrobial activity against these bacterial isolates.
In current study 70 throat swab samples were collected from Children Hospital, Lahore and processed to isolate and identify antibiotic resistant sore throat bacteria by swabbing on blood agar andTryptic soya agar.Initial identification was done using conventional biochemical tests and confirmation was done through API 20 Strep. Antibiotic resistance pattern in isolated bacteria was done by modified Kirby Bauer disc diffusionas per CLSI,2014 criteria.Extracts (Ethanolic, boiling and distilled water) of herbs (Liquorice, ginger, cinnamon, red dates and kalonji) was obtained and tested for antimicrobial activity against resistant isolates. Efficacy of the herbal extracts was evaluated through Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentrations. Ethanolic extracts of Liquorice, ginger, cinnamon and red dates have good antimicrobial activity against resistant sore throat bacterial isolates than boiling and distilled water extracts.
The present study demonstrated that S. pyogenes and Staphylococcus are major bacteria responsible for sore throat infection in children. These isolates have variety of different antibiotic resistance patterns. In order to minimize emergence of antibiotic resistant isolates, herbal extracts can be used.Ethanolic extracts of liquorice, ginger, cinnamon, red dates have good antibacterial activity as compared to kalonji. Statistical analysis showed that ethanolic extracts of Liquorice has a significant activity with ginger, cinnamon and red dates.The degree of antibacterial activity of plant extracts testedcan be graded in the following order: Liquorice> ginger>Cinnamon>red dates>kalonji. Itis estimated that by using natural products will reduce the resistance in microorganism.
Statistical analysis:
The data collected in MS Excel 2016 and analyzed statistically by one way Analysis of Variance using Statistical Package for Social Science (SPSS) 18.0 software.
Availability: Items available for loan: UVAS Library [Call number: 2488-T] (1).
5.
Studies On Biological Control Of Salmonellosis In Poultry
by Kiran Imtiaz (2010-VA-296) | Dr. Ali Ahmad Sheikh | Prof. Dr. Masood Rabbani | Dr. Wasim Shehzad.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Contaminated poultry food products are the main cause of Salmonellosis all over the
world. Salmonella may enter in different poultry farms through vertical or horizontal
transmission. Previously, Salmonella infection and number were controlled in poultry and
their products by using different methods such as usage of hazard analysis critical control
point (HACCP), antibiotics, symbiotics, etc. But not any method gives the better result that
shows the reduction of Salmonella species in poultry that cause infection. Therefore, the
attention was diverted to find the alternative therapies such as the usage of bacteriophage,
Bacteriophage specific to Salmonella used in reduction of Salmonella number as a biocontrol
agent in poultry origin. It proves effective for Salmonella reduction.
In the current study samples for isolation of Salmonella and bacteriophages were
collected. Salmonella was isolated from liver, caeca and lungs of infected chickens collected
from different infected poultry farms. After processing of sample according to the literature
method, confirmation of salmonella from samples were done by inoculating them separately
on S.S agar by streaking method. Further confirmation of Salmonella was done on the basis
of biochemical testing.
Bacteriophage was isolated from sewage water near to the poultry farms. For their
isolation, sewage sample was centrifuged and then supernatant was collected in separate tube.
Filtration was done of Supernatant, and then Filtrate was used as a bacteriophage source.
Bacteriophages in filtrate were confirmed by spot test method. After confirmation of both, in
vivo study was performed to evaluate the effect of bacteriophage on Salmonella when
inoculated in combination or separately in chickens groups.
51
Summary
Statistical analysis:
The data will be transferred on spreadsheet using MS Excel 2010. The data will be
analyzed through one way ANOVA test using Statistical Package for Social Sciences (SPSS)
version 18.0.
The present study was helpful to determine the effect of bacteriophage in reduction of
Salmonella in commercial broilers sector. Antibiotic resistant Salmonella infection is difficult
to control using conventional ways of antibiotic therapy and is responsible for huge economic
losses in poultry. Chicken groups that were used for invivo study showed that bacteriophage
was proved very effective in reduction of Salmonella either gives it in combination with
Salmonella or separately to poultry. It is predicted that bacteriophage therapy is better than
the conventional ways for reduction of Salmonella infection in poultry sector. It is essential
that the research should be continue to study the effect of bacteriophage in reduction of
specie specific Salmonella, and to determine the effect of different physicochemical factor on
their activity. Availability: Items available for loan: UVAS Library [Call number: 2511-T] (1).
6.
Evaluation Of Inhibition Activity Of Indigenous Lactobacilli Spp. On Ammonia Emitting Bacteria
by Fatima Sajjad (2010-VA-312) | Dr. Muhammad Nawaz | Prof. Dr. Masood Rabbani | Dr. Sehrish Firyal.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Poultry is the 2nd largest industry in Pakistan which is growing at an amazing rate of more
than 10% from last few years. It provides jobs for more than 1.5 Million people. Although, it is
growing at excellent pace, it still faces many problems. One of the important problems in poultry
farming is the production of ammonia by urease producing microbes. Ammonia is health hazard
for both poultry and human. Urease producing bacteria which are the major problems in poultry
are Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae and Staphylococcus
aureus in poultry droppings. Probiotics such as Lactobacilli can inhibit the urease producing
bacteria in poultry intestine as well as environment thus mitigate the ammonia emission. Present
study was designed to isolate Lactobacilli from indigenous poultry, screen them for
antimicrobial activity against urease producing microbes and determine their effect on growth of
Proteus mirabilis during co-culture experiments. A total of 71 Lactobacilli isolated were
recovered from 20 samples of droppings (10) and caeca (10) of back-yard poultry on MRS agar
plates. Twenty seven (27) isolates demonstrated antimicrobial activity against ammonia emitting
bacteria by agar spot and well diffusion assays. Seven isolates (FSL19, FSL25, FSL39, FSL45,
FSL51, FSL63 and FSL71), having better antimicrobial activity, were selected for co-culturing
with Proteus mirabilis in nutrient broth. FSL25, FSL45, and FSL51 showed more than 2 log10
reduction of Proteus mirabilis in co-culture experiments. FSL25, FSL45, and FSL51 were
identified as Lactobacillus plantarum, Lactobacillus fermentum and Lactobacillus salivarius,
respectively by amplifying and sequencing their partial 16S rDNA. It is concluded that
Lactobacillus plantarum FSL25, Lactobacillus fermentum FSL45 and Lactobacillus salivarius FSL51 may be used to mitigate ammonia emitting bacteria in poultry environment after further
investigations. Availability: Items available for loan: UVAS Library [Call number: 2594-T] (1).
7.
Identification of Ciprofloxacin Resistant Bacteria of Public Health Significance in Animals and Poultry
by Qurat ul Ain (2010-VA-306) | Dr. Arfan Ahmad | Prof. Dr. Masood Rabbani | Dr. Muhammad Nasir.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: In the present study ciprofloxacin resistant isolates with public health significance, collected from chicken five each of respiratory, digestive and reproductive system where as twenty samples each of respiratory, digestive, reproductive and milk from, goat, sheep, cow and buffalo has been determined. Out of 95 samples collected, 52 samples were identified as ciprofloxacin resistant bacteria which were E. coli, Salmonella, Staphylococcus, Pseudomonas, Klebsiella and Proteus from chicken (respiratory, digestive, reproductive system) and goat ,sheep, cow and buffalo( respiratory, digestive, reproductive system and milk).
In chicken out of 15 samples, respiratory bacterial isolates showed maximum resistance (80%, 4/5) against ciprofloxacin as compared to those from digestive (60%, 3/5) and reproductive system (40%, 2/5). In goat out of 20 samples, respiratory bacterial isolates showed maximum resistance (80%, 4/5) against ciprofloxacin as compared to those from digestive (20%, 1/5), reproductive system (60%, 3/5) and milk (40%,2/5). In sheep out of 20 samples, digestive bacterial isolates showed maximum resistance (60%, 3/5) against ciprofloxacin as compared to those from respiratory (40%, 2/5), reproductive system (20%, 1/5) and milk (40%,2/5).
In cow out of 20 samples, digestive bacterial isolates showed maximum resistance (100%, 5/5) against ciprofloxacin as compared to those from respiratory (80%, 4/5), reproductive system (60%, 3/5) and milk (20%,1/5). In buffalo out of 20 samples, bacterial isolates in milk samples showed maximum resistance (80%, 4/5) against ciprofloxacin as compared to those from respiratory (60%, 3/5), reproductive (60%, 3/5) and digestive system(40%, 2/5).
Summary
61
In Pakistan, there is also increasing trend of emergence of antibiotic resistant bacteria in human infection. In our study from various systems, ciprofloxacin resistant bacteria were isolated and confirmed. Among these bacteria irrespective of animal spp and systems, E. coli was found most abundantly in respiratory system of chicken and digestive system of cow due to irrational use of antibiotics and simultaneously use in animals, poultry, and human . In this study comparative status of ciprofloxacin resistant bacteria such as E. coli, Salmonellae, Klebsiella, Staphylococcus, Pseudomonas and Proteus with public health significance from chicken and animals and their associated risk factors were observed. Availability: Items available for loan: UVAS Library [Call number: 2622-T] (1).
8.
Relationship Of Parent’s Level Of Education And Socio-Demographic Variables With Child’s Immunization Status: A Cross-Sectional Study In The Rural Areas Of District Layyah
by Abida Zahoor (2012-VA-573) | Prof. Dr. Mansur-ud-Din Ahmad | Prof. Dr. Muhammad Athar Khan | Prof. Dr. Masood Rabbani.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Immunization is a very important element of public health. It is the process whereby a person is made immune or resistant to an infectious disease, generally by the administration of a vaccine. Vaccines stimulate the body’s own immune system to protect the person against subsequent infections. It prevents against various communicable diseases such as Tuberculosis, Tetanus, Pertussis, Diphtheria, Poliomyelitis, Hepatitis B and Measles.
A cross-sectional study was conducted on 200 parents having at-least one child under five years of age. Data was collected from the rural areas of district Layyah. The purpose of the study was to investigate the relationship between parent’s level of education and child’s immunization status on one hand and the effect of sociodemographic variables on child’s immunization status on the other hand. In this study, it was concluded that there was a significant association between parent’s level of education and the socioeconomic variables with child’s immunization status.
Low rate of parent’s literacy, especially health literacy, poor socioeconomic status, large population size, parent’s refusal, difficulty in accessing immunization services and lack of health facilities were identified as the main barriers to immunization completion. As parent’s level of education is considered to be the corner stone in the progress of modern nation so education is a very important element as educated parents play a significant role in achieving the health of their children. It was concluded that there was a significant association between literacy status of parents, income status of parents and the immunization status of children. The immunization status of children in the rural areas can be improved by higher household income, literacy, better
Summary
41
health knowledge, exposure to media, maternal empowerment and mother’s participation in decision-making process. The policy makers should stress on the education of the people especially female education. Health awareness campaigns should be carried out so that parents can gain the benefits of vaccination.
6.1 Hypothesis
Ho: there is no significant relationship between parent’s level of education and child’s immunization status.
H1: Parent’s level of education has significant relationship on child’s immunization status
6.2 Methodology A cross-sectional study of six months duration from July 2016 to December 2016 was conducted in the rural areas of district Layyah in which parents of children under 5 years of age were interviewed about the immunization status of their children. Convenient sampling technique was used to collect the data. Data was collected from 200 parents living in the rural areas of district Layyah. The collected data was analyzed by using SPSS version 16.0.
6.3 Statistical Design
The dependent variable “child’s vaccination” and independent variables “parent’s educational level, household income, parent’s occupation and family composition” were analyzed by using SPSS version 16 and Microsoft Excel. Data entry and analysis was done on SPSS-16. Chi-square test was used for statistical testing. A p-value of <0.05 was considered significant. Frequencies and percentages of categorical variables were calculated. Cross
Summary
42
tabulations were done among dependent and independent variables. Chi-square test was applied on different sociodemographic factors and child’s immunization status to define the significant associations.
6.4 Study Outcomes It is concluded that there is a significant association between literacy status of parents, income status of parents and the immunization status of children. The major benefit of this study is that, the research findings can be used to assess the relationship of parent’s education with child’s immunization status and to find out those factors which are the main hindrance in child’s vaccination coverage. It will also provide new dimensions for further research related to child immunization and the health managers will find new facts and figures that will help them to make national decisions with the certainty of success. Availability: Items available for loan: UVAS Library [Call number: 2654-T] (1).
9.
Isolation Of E.Coli O157: H7 From Cattle And Buffalo Meat From Slaughterhouse Of Two Different Management Systems In Lahore
by Ali Hassan Rehman (2008-VA-144) | Dr. Aamir Ghafoor | Prof. Dr. Masood Rabbani | Dr. Hassan Mushtaq.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The present study was designed to identify the presence of E. coli O157:H7 in buffaloes and cattle carcasses that were slaughtered in two different management systems in Lahore. In Management system A, proper risk assessment and hazard analysis was done and food safety protocols were followed in contrast to Management system B in which animals slaughtered on ground and no hygienic practices were followed. The present study was attempted to detect presence of E. coli O157:H7 in carcasses that were slaughtered in these management systems. For the confirmation of whether E. coli O157:H7 is present or not serological test were performed. For this Latex agglutination Kit method was used.
A total of 100 meat samples, in which 50 (25 Cattle and 25 Buffaloes) were collected from management system A and 50 (25 Cattle and 25 Buffaloes) were collected from management system B. These samples were pre-enriched with Modified E. coliBrothand afterward cultured on Cefixime - tellurite Sorbitol MacCkony Agar. The colonies which were colorless and small were selected for confirmation with Latex agglutination test.
For confirmation of E. coli O157 latex agglutination kit reagentswere allowed to come to room temperature. A pipette was used to transfer 0.2 ml normal saline into a 12 x 75-mm test tube.Using a sterile loop or needle, pick sufficient colonies from the plate and suspend them in the saline to achieve turbidity. One drop of the Prolex™ E. coli O157 Latex Reagent(Blue color) was placed in a test circle on the test card. Using a Pasteur pipette one drop of the test suspension was added into the same test circle and mixed by using stick provided. The card was gently rocked and examined for agglutination for up to two minutes. Isolates that were positive result with the test latex were tested further by repeating the procedure using the Prolex™ Negative Control Latex Reagent.
The positive samples that showed agglutination were declared as positive for E. coli O157:H7.
1 cattle was positive for E. coli O157:H7 in management system A and 4 Cattle were positive in management system B, no buffalo meat sample positive in both systems. The results showed that E. coli O157:H7 was present in our meat value chain and a food safety hazard also. In management system A 1 meat sample was positive for harmful pathogenic bacteria. In management system B, 4 meat samples were positive for E. coli O157:H7.
The conclusion of this study, Management system A is better than Management system B to minimize the occurrence of E. coli O157:H7.
Availability: Items available for loan: UVAS Library [Call number: 2647-T] (1).
10.
Prevalence And Chemotherapy Of Dry Cow Mastitis
by Abdul Sattar Saqib (2014-VA-766) | Dr. Muhammad Avais | Prof. Dr. Aneela Zameer Durrani | Prof. Dr. Masood Rabbani.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The study was undertaken to determine the prevalence and chemotherapy of dry cow mastitis. Mastitis is responsible for a wide range of health problems and economic losses in cows and is characterized by decrease in milk production, Swelling of the udder, hotness of the udder and anorexia. All lactating animals generally have a period of 6-10 weeks preceding to calving (usually annually) as a dry or resting period, a non-lactating phase. About to calving the cow remains at risk to new intra-mammary infections, especially shortly after the ‘drying off’ or termination of milking. During the dry period the prophylactic benefit of 82% reduction in the rate of intra-mammary infection is the result of the dry cow treatment with antibiotics and higher rate of eliminating infections than treating in lactation.
For this purpose, 250 Pregnant dry cows were examined for subclinical mastitis. The milk samples were collected from Pattoki and adjacent areas and California mastitis test (CMT) was performed and positive samples were furtherly processed for somatic cell count at medicine Laboratory of University of Veterinary and Animal Sciences, Lahore.
For Chemotherapy, 24 animals positive for dry cow mastitis were equally divided into 4 groups viz A, B, C and D. Each group comprising of 6 animals. The animals of group A were treated with intramammary antibiotic Cloxacilline+ Ampicillin (Masticlox ,ICI). Animals in group B were treated by injecting 2 shorts (72 hours interval) of long acting Amoxicilline (amoxy 150 L.A Floris veterinaire produkten B.V Vught the Netherland) intra muscularly. Animals in group C were treated with Cephradine (Velosef, GSK) 1g/quarter through intramammary route once. Cows in group D were served as positive control. Animals in all groups were kept under close observation for clinical mastitis until parturition. After calving, cows in each group were tested for mastitis at days 7, 14 and 21 (post calving) using CMT and
Summary
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SCC. Effectiveness of a particular treatment was determined on the basis of CMT score and SCC.
The collected samples from Pattoki and adjacent areas were processed at the Medicine laboratory at UVAS, Lahore aseptically for CMT and CMT positive samples were processed by Somatic cell count (SCC).
Overall prevalence determined which was 39.60% (99/250samples) by CMT and SCC.The efficacy of different antibiotics used in chemotherapy of dry cow mastitis was checked. The efficacy of Amoxicilline (Amoxy 150 L.A), Ampicillin+ Cloxacilline (Masticlox) and Cephradine (Velocef 1g) was recorded at day 7,14 and 21days post calving.
Group A was treated with Ampicillin +Cloxacilline (Masticlox) its efficacy was 83.33% and group B was treated with Amoxicilline (Amoxy 150 L.A) and its efficacy was 66.66% effective while the efficacy of Cephradine (Velocef 1g) was 33.33% in group C.
From this study it was concluded that CMT is more reliable test than other tests for the diagnosis of Mastitis in cows. Secondly subclinical mastitis which is an important problem of cows is significantly prevalent in dry cows in Pattoki and adjacent areas. Cloxacilline+ Ampicillin and Amoxicillin is the most effective drug while Cephradine is relatively less effective against mastitis in dry cows. Availability: Items available for loan: UVAS Library [Call number: 2669-T] (1).
11.
Antibacterial Activity Of Various Plant Extracts Against Waterborne Bacteria
by Anam Munir (2010-VA-292) | Dr.Fareeha Akhtar | Prof. Dr. Masood Rabbani | Dr. Raheela Akhtar.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Pure water is basic necessity for not only human being but also for animals. With increasing population, it becomes very hard to provide safe and pure water to the community. Due to unavailability of proper swage system, portable water is contaminated in many ways. Poor water quality reported in 2004, hygiene and sanitation account for some 1.7 million deaths a year (Ashbolt 2004). It is dangerous for health so there is need to disinfect the water. Water is not disinfected in most of rural areas in Pakistan. Disinfection of water is necessary for health and reducing the chances of spread of diseases through water.
Expensive and sophisticated methods e.g. chemical treatments are not easily available at rural areas. Chemical methods are not only costly but also leave harmful DBP (disinfection by products) in water. Recent researches have proved that DBP(s) are carcinogenic and also cause neurological problems. To overcome the problem of contamination and DBP, herbal extracts are good choice for this purpose as they are eco-friendly, cheap and easily available in rural areas. They have well known antibacterial activity. To overcome the problem of waterborne bacterial pathogens, various herbal extracts are used to disinfect the water. These herbs are easily available in rural areas and can disinfect the water.
In current study, 50 water samples from different rural areas of Lahore were collected. Water samples were processed in 24 hours after collection to evaluate the bacterial load in water along with it water was also given treatment with herbal extracts at the same time.
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Bacterial load was evaluated by technique of total viable count (TVC) and most probable number (MPN) for coliforms. Evaluation of bacterial load and treatment with herbal extracts was done on same time so that efficacy of extracts to reduce the number of bacteria in water could be examined. Comparison of bacterial load before treatment and after treatment gave the idea of efficacy of antibacterial activity of herbal extracts.
Herbal extracts (ethanolic, fresh juice, aqueous) of herbs (Neem, Tulsi) and seeds of kalonji were made. These extracts were tested for antimicrobial activity against bacteria and coliform present in samples of water. Treatment with herbal extracts was given to water in ratio of 1:1. After 24 hours, the treated water was evaluated for reduction of bacterial load by TVC. Ethanolic extracts of Tulsi as well as Neem had good results than aqueous and fresh juice extracts.
The present study demonstrated that contamination of water is major issue of Pakistan. It results in great economic loss of livestock and poultry section. Chemical disinfection is dangerous for health. So there is need to disinfect water by safe and ecofriendly way. Herbal extracts are good choice for this purpose. Ethanolic extracts of Neem has good antibacterial activity than Tulsi and Kalonji. Statistical analysis showed that ethanolic extracts of Neem has more significant activity than Tulsi and Kalonji. The degree of antibacterial activity of plant extracts for Neem, Kalonji and Tulsi can be graded on the base 0f diluent used. There is no significant difference between antibacterial activities of all three herbs. Tulsi, Neem and kalonji have equally effective antibacterial activity. Whereas ethanolic extract showed more effective
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results than aqueous and fresh juice extract. It is estimated that by using natural herbs and seeds of plants, contaminated water can be bacteria.
Statistical analysis:
The data collected in MS Excel 2016 and analyzed statistically by univariate analysis of variance using the Statistical Package for Social Science (SPSS) 16.0 software. Availability: Items available for loan: UVAS Library [Call number: 2673-T] (1).
12.
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).
13.
In Vitro Biological Control Of Avian Pathogenic Escherichia Coli
by Aleena kokab (2011-VA-418) | Dr. Ali Ahmad Sheikh | Prof. Dr. Masood Rabbani | Dr. Waseem Shehzad.
Material type: Publisher: 2017Dissertation note: Avian pathogenic E. coli (APEC) has meticulous virulence properties making it a
potential pathogen causing insidious infections in poultry, termed as colibacillosis either
as primary pathogen or as a secondary pathogen. All over the world, there are major
economic losses in poultry industry due to this disease. To prevent APEC infection,
strategies include improving hygienic methods, vaccination, antibiotic treatment and
introduction of novel immunopotentiators but all these measures had limited success.
Moreover, because of extensive use of antibiotics for treatment purposes, antibiotic
resistance is major growing concern. Bacteriophage therapy is promising new alternative
to antibiotics. There are many benefits of phages over antibiotics and are now used in
research for the treatment of enteric and respiratory problems in poultry and are proved to
be effective in reducing E.coli infection.
In present study bacteriophages was used to minimize and control the number of
E.coli causing colibacillosis in poultry. For this, E.coli (n = 10) from liver of infected
commercial poultry birds and E.coli (n = 5) from intestines of apparently healthy birds
was isolated and characterized using biochemical tests. Final confirmation of E.coli
isolates was done by PCR. To differentiate between pathogenic and non-pathogenic
E.coli, Congo Red Dye binding test (CR test) was performed. Antibiotic resistance
profiling of the isolates was also determined against 10 commonly used antibiotics
through Kirby-Bauer disk diffusion method.
For the isolation of bacteriophages, sewage or poultry sludge sample was
processed according to literature method and purified bacteriophages (n = 5) were
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assessed for their stability at various temperature and pH. Bacteriophages were also
assessed for their In-Vitro Lytic Activity of against E. coli isolates.
Five numbers of lytic bacteriophages were isolated and purified against avian
pathogenic E.coli, showing resistance against Ampicillin, Doxycycline, Ciprofloxacin,
Neomycin, Trimethoprim-sulphamethoxazole, Nalidixic acid and Ceftriaxone. Isolates
were also classified into pathogenic and non-pathogenic on the basis of their ability to
bind Congo red dye on Congo red media. Phages were highly lytic against pathogenic
isolates while there was no lytic activity against non-pathogenic isolates thus making
them unique for in vivo trials. Phages were found to be stable at temperature from 25oC
to 56oC with highest stability at 37oC while on the basis of pH, phages were stable
between 5-9 pH with highest stability at pH 7. Maximum lytic activity observed was for
12 hours and then there was emergence of resistant bacteria and this problem may be
tackled by increasing MOI or by using combinations of phages against resistant bacteria.
Present study helped to reveal the effectiveness of bacteriophages against antibiotic
resistant E.coli and proved that bacteriophages can be used as promising alternatives to
antibiotics for reducing E.coli infection in poultry. Availability: Items available for loan: UVAS Library [Call number: 2829-T] (1).
14.
Characterization And Physicochemical Optimization Of Phytases Produced By Indigenous Isolates Of Lactobacillus SPP.
by Aanisa Arif (2011-VA-424) | Dr. Muhammad Nawaz | Prof. Dr. Masood Rabbani | Dr. Sanaullah Iqbal.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Phytate is one of the major organic storage forms of phosphorous of phosphorus in
seeds, cereal, oil and legume; in nature about 75%-80% of total phosphorus is available in
this form. Phosphorus is stored in roots and in seeds and cereals as phytate. Phytases are
responsible for breakdown of phytic acid (phytate) into inorganic monophosphates and
free myo- inositol. Phytases are a class of phophatases which hydrolyze phytic acid into
inorganic phosphate and myo inositol or less phosphorylated phosphates. Monogasteric
animal like poultry, human and fish lack phytase due to which they cannot derive
phosphate from phytate and phosphorus is unavailable to them.
So, present study is designed as a first step in a multi-step project to develop
indigenous phytase producing probiotic lactobacilli from different sources, the
optimization of phytase production and effect of physical & chemical factors on the
phytase stability and activity. Lactobacillus isolated from poultry was checked for phytase
production on Phytase screening media (PSM). Enzyme from the isolates showing activity
were quantified by ammonium molybadate method, the enzyme production were
optimized at different physical and chemical parameters such as temperature (30, 35 &
42°C), pH (4,5,6,7 & 8), osmotic pressure (1%,2% and 4%), aerobic/anaerobic conditions,
carbon (glucose, lactose, sucrose), nitrogen sources (peptone, tryptone & urea) and bile
salts (0.3%,1% and 2). Enzyme was partially purified and characterized by SDS-PAGE.
In present study 20 samples of poultry droppings (SP01-SP10) and fermented food (SY01-
SY10) were processed for isolation of lactobacilli. A total of 90 isolates (PDP01-PDP45;
FYP01-FYP45) were selected from MRS plates. Isolates were preliminary confirmed as
Gram positive rods with Catalase negative. All isolates were further purified and stored in
MRS broth supplemented with 15% glycerol at -20oC. Purified lactobacilli isolates were
screened for phytase production on phytase screening medium and zone o f hydrolysis was
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measured in mm. Out of total of 90 isolates 62 isolates showed phytate hydrolysis. Out of
62 isolates, 16 were selected on the basis of retention of hydrolysis zone after cobalt
chloride staining. Out of 16 selected isolates, eight isolates PDP05, PDP09, PDP10,
PDP16, PDP23,PDP24, PDP30 and PDP35 were of poultry origin and eight FYP12,
FYP15, FYP17, FYP21, FYP26, FYP31, FYP38 and FYP42 were of fermented foods.
Selected isolates and retention of their zone of hydrolysis after cobalt chloride staining are
given in table 4.4. Phytase activity of selected lactobacilli isolates was checked in
modified MRS broth containing 0.2% sodium phytate at 37°C after 24 hrs. Cell free
supernatant was used as crude source of enzyme. Enzyme activity was determined by
using ammonium molybadate method. Amplification and sequencing of 16Sr DNA
(≈1500bp) was done by using universal primers which revealed PDP10, PDP24 and
FYP38 had >99% similarity with Lactobacillus gallinarum, Lactobacillus reutri and
Lactobacillus fermentum respectively with GenBank accession no. MF980924,
MF980925 and MF980923 respectively.
Phytase production by lactobacilli was optimized at different parameters e.g.
temperature (30, 35 and 42°C), pH (4, 5, 6, 7 and 8), osmotic pressure 1%, 2% and 4%.
The effect of oxygen was determined by growing lactobacilli isolates in aerobic and
anaerobic conditions followed by measuring enzyme activity. PDP10, PDP24 and FYP38
showed the best activity at 35°C (6.86 ± 0.15 IU/ml, 5.12 ± 0.12 IU/ml and 5.65 ± 0.13
IU/ml respectively) at pH 5 (6.86 ± 0.15 IU/ml, 5.12 ± 0.12IU/ml and 5.50 ± 0.13 IU/ml
respectively). Maximum phytase activity was recorded at 1% NaCl 4.78 ± 0.14, 4.18 ±
0.13 and 5.58 ± 0.12 IU/ml respectively) whereas anaerobic conditions were favourable for
the production of phytase by selected isolates.
Effect of carbon, nitrogen sources and bile salts was determined by growing
isolates MRS broth (0.2% sodium phytate) modified with different carbon (glucose,
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lactose, sucrose) and nitrogen sources (peptone, tryptone and urea) and bile salts (0.3% ,
1% and 2%) followed by measuring enzyme activity. In this study isolates PDP10, PDP24
and FYP38 exhibited maximum phytase activity in the presence of 2% glucose as
compared to other carbon sources lactose and sucrose (4.36 ± 0.11, 4.38 ± 0.18 and 5.01 ±
0.15 IU/ml respectively). Present study revealed 0.1 % peptone as an optimal source of
nitrogen for PDP10 and PDP24 (4.54 ± 0.13 and 4.23 ± 0.19 IU/ml respectively) while
FYP38 (4.56 ± 0.14 IU/ml) showed best result in presence of 0.1% tryptone. All the
isolates showed maximum phytase activity at 0.3% bile salt concentration as compared to
1% and 2% concentration. Enzyme activity of phytase obtained from PDP10 was not
varied while treating it at different pH (4, 5, 6, 7 and 8) at different intervals of time.
Enzyme activity of phytase obtained from PDP24 was lower at pH 8 for 15, 30, 45 and
60min (3.41± 0.10, 3.40 ± 0.09, 3.42 ± 0.08 and 3.41 ± 0.11IU/ml respectively). Enzyme
activity of phytase obtained from FYP38 was lower down from pH 7 to 8 for 15, 30, 45
and 60min (4.41 ± 0.09, 4.42 ± 0.11, 4.43 ± 0.10, 4.41 ± 0.12 IU/ml respectively) & (4.40
± 0.09, 4.31 ± 0.11, 4.33 ±0.10 and 4.34 ± 0.12 respectively). Enzyme activity was
inhibited at 1mM and 5mM concentrations of Ca2+ while metal ions like Mg2+ and Na2+
addition stimulated the phytase activity. Approximate molecular weight of extracellular
protein precipitated from the cell free supernatant of PDP10, PDP24 and FYP38 was
~50kDa. Availability: Items available for loan: UVAS Library [Call number: 2895-T] (1).
