1.
Staphylococcal Coagglutination Test For Rapid Detection Of Foot And Moth Disease Virus
by Baitullah Khan | Dr. Atif Hanif | Prof. Dr. Masood Rabbani | Faculty of Veterinary Sciences.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2010Dissertation note: Foot and mouth disease is a highly contagious, viral disease of cloven hoofed animals and causes high economic losses. Rapid detection of FMD is necessary to control the disease from spreading. Although several reliable tests like ELISA, CFT' and PCR already exit but none of them applicable in field conditions. The aim of this study was to optimize rapid, economical and sensitive test for the detection of FMD. For this purpose rabbits were used to raise immune sera against FMD virus with one uninnoculated control. Immune sera collected from these rabbits at different time interval and presence of antibody was determined by using AGPT. Immune sera were then conjugated with satbalized and inactivated staphylococcus aureus cell using different dilutions. Cell wall of S. aureus contains Protein A which naturally binds with Fc portion of IgG leaving the Fab portion to interact with antigen. In presence of homologues antigen causing agglutination was seen with nacked eyes. Light blue background was found best while observing results.
The coagglutination test was applied on FMD known antigen. Clear agglutination on slide was observed by mixing equal quantity of COAT reagent and its respective antigen. Total 40 vesicular fluid samples from FMD infected animals were tested with COAT, in which 38 yielded positive results and the remaining two yielded negative results. COAT reagents were also tested against PPR virus depicting negative results. COAT was found specific for FMD antigens. This test is quick and generates results within five minutes.
This reagents of CAOT also applied on two fold dilution of vesicular fluid from FMD infected animal and positive result were observed up to 1:32 dilution. This test is sensitive, specific, economical and rapid for detection of FMD. This test was successfully used for detection of FMD in filed.
Availability: Items available for loan: UVAS Library [Call number: 1176,T] (1).
2.
Development And Optimization Of Multiplex Pcr For The Identification Of A, O And Asia 1 Strains Of FMDV In Pakistan
by Muhammad Ikram | Dr. Atif Hanif | Dr. Imran Najeeb | Prof. Dr.
Material type: ; Format:
print
Publisher: 2010Dissertation note: Foot and mouth disease (FMD) is highly infectious disease of cattle, buffalo, sheep and goats. It is caused by genus Aphthovirus of Picomaviradae family. FMDV is RNA virus having seven serotypes A, 0, C, Asia I, SAT1, SAT2 and SAT3.
Foot and mouth disease is endemic in Pakistan and causes high economic losses to livestock industry. So priority is to develop quick and efficient methods for detection of FMDV and to limit the spread of disease outbreak. Although CFT, VNT and ELISA are already being used for the diagnosis of FMDV in Pakistan but these diagnostic techniques are time consuming and their specificity and sensitivity is low. Multiplex PCR for the identification of FMDV is very much sensitive and specific, can be done with in three hours after the receipt of samples.
Present study has been designed to optimize multiplex RT-PCR for rapid detection of FMD virus. RNA was extracted from virus stock obtained from QOL, UVAS Lahore and from field samples. After RNA extraction the samples were subjected to synthesize cDNA by the use of Reverse Transcriptase enzyme. After cDNA synthesis PCR reaction was carried out. The amplified products were resolved on 1.5% Agarose Gel. A multiplex RT-PCR strategy was optimized and developed for the detection of virus serotypes A, 0 and Asia l.
Restulst of this study helped to develop an efficient and economical method for rapid detection of FMD virus and also helpful in differential diagnosis from other vesicular diseases.
Availability: Items available for loan: UVAS Library [Call number: 1189,T] (1).
3.
Preparatuin And Evaluation Of Monospecific Anisera Against Hemagglutinin, Neuraminidase And Matrix Proteins of local Avian Influenza Strains H5 N1, H7 N3, H9 N2, for diagnostics
by Sumaira Ijaz | Dr. Atif Hanif | Dr. Aftab Ahmad Anjum | Prof. Dr.
Material type: ; Format:
print
Publisher: 2011Dissertation note: Avian Infuenza is an economically important disease of poultry worldwide. It has caused losses to poultry industry on larger scale. It is important due to its zoonotic nature. Studies were carried out to raise monospecific antisera against hemagglutinin, neuraminidase and matrix antigens. HA, NA and M proteins of each of the avian influenza strains were separated on Sodium Dodecyl Sulfate Poly Acrylamide Gel Electrophoresis (SDS PAGE). First virus was lysed to release the proteins. Virus was lysed by using 4% Triton X 100, 1mM KCl and 0.01M Tris buffer. Then the sample was dialyzed. Sample was run on gel to purify proteins. The protein bands of appropriate molecular weight were cut and triturated in 1ml of normal saline. Material was centrifuged to remove the gel content.
Each protein was confirmed by the Bradford's Reagent. Each protein was individually mixed with Montanide ISA 50 adjuvant in 1:1 ratio to make the vaccine. Vaccine of each polypeptide of AIV strains was injected in three groups of nine birds each. One group of birds was injected with HA, second group with NA and third group with Matrix proteins of H?, H? and H?. Three groups of birds served as control. The blood samples of all birds were collected before and after inoculating vaccine. The sera of birds before and after inoculating vaccine were checked for antibodies titre against HA antigen by HI test. Antibodies against Matrix antigen were detected by Agar Gel Precipitation Test. Antibodies titre was raised after inoculating polypeptides. In case of sudden outbreaks, antisera may be helpful to control disease.
Availability: Items available for loan: UVAS Library [Call number: 1351,T] (1).
4.
Molecular Characterisation And Antibiotic Resistance Of Avian Salmonella Enterica
by Sajid ul Hassan Qureshi | Dr. Atif Hanif | Prof. Dr.Masroor Elahi Babar.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Avian Salmonella enteric sub species enteritidis can cause enteritis in a wide range of host species and being responsible for the majority of Salmonella food-borne enteritis in man worldwide. This food borne disease of primary concern in developed, as well as developing countries. The spread of disease is favored by a variety of animal reservoirs and a wide commercial distribution of both animals and food products. This disease is among one of the major public health problems in terms of socio-economic impact. However there was little information on their prevalence, characterization and antibiotic susceptibility in Pakistan. In the current study Salmonella enteritidis was characterized on the basis plasmid encoded antibiotic resistance and Whole cell protein profiling to reduce the public associated with consumption of infected products. Ten Isolates were tested for plasmid encoded antibiotic resistance against six antibiotics. results of in vitro susceptibility by standard disc showed that all isolates were highly resistant to Ampicillin (100%), followed by Amoxicillin to which 60% isolates were resistant. High susceptibility was shown to levofloxacin, and Ciprofloxacin (90 and 80% respectively) by S. enteritidis isolates. Plasmid profiling of resistant isolates shown a large plasmid ( kb) that appears to be a serotype specific virulence plasmid. Whole cell protein profiling was done by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) which revealed results close relation among Salmonella entertidis isolates and the 78.1 and 35 kDa bands were observed to be major bands in all strains.
Availability: Items available for loan: UVAS Library [Call number: 1380,T] (1).
5.
Molecular Characterisation And Antimicrobial Resistance Of Human Salmonella Enterica
by Yaser Bhatti | Dr. Atif Hanif | Prof. Dr.Masroor Elahi Babar.
Material type: ; Format:
print
Publisher: 2012Dissertation note: Infectious diseases directly or indirectly are still the main cause of morbidity and mortality in the human population. Prevention and control of such diseases has been a major challenge since ages. During the last few decades, infections with Salmonella have been recognized as a major hazard to humans in most countries.In this study, ten confirmed and purifiedisolates ofSalmonella typhi were collected from Chugtai's Lahore Lab.and were grown inbuffered peptone water, tetrathionate broth and SS agar. Isolates were characterized by antibiotic resistance patterns, plasmid profiling and protein profiling. All isolates were resistant to ampicillin and were sensitive to ciprofloxacin and levofloxacin. 90% of isolates were sensitive to gentamicin and 50% were resistant to chloramhenicol, while 70% of isolates showed intermediate behaviour to amoxicillin. Single plasmid profile was observed among all resistant isolates and all isolates harboured a single heavy weight plasmid.Sensitive isolates were free of any plasmid. Isolates were grouped into six groups by SDS-PAGE. Different banding pattern was observed among groups. However, a protein of 10kDa was common in all isolates.
Availability: Items available for loan: UVAS Library [Call number: 1382,T] (1).
6.
Alayisis Of Sodium Channel Subunit Beta-1 ( Scnib ) Mutations Involved In Generalized Epilepsy With Febrile Seizures
by Salma siddique | Dr.Muhammad Wasim | Dr. Abu saeed hashmi | Dr. Atif Hanif.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2011Dissertation note: Epilepsy is, one of the most common disorders of the brain, a common neurological
condition defined by recurrent and unprovoked seizures. Epilepsy affects 50 million
people worldwide, including one in every 200 children. Febrile Seizures (FS) are not
thought of as a true epileptic disease but rather as a special syndrome characterized by its provoking factor (high grade fever) and a typical range of 6 months to 6 years. The
patients with generalized epilepsy with febrile seizure plus (GEFS+) show febrile
seizures initially, lasting beyond 6 years of age, and afebrile seizures occur with multiple
types, mainly with generalized seizures but sometimes with focal seizures. Studies have
shown that genetic factors play an important role in the pathogenesis of GEFS+ and other types of epilepsy. Mutations in a number of genes have been identified that leads to the various types of epilepsy. Sodium channel subunit beta-l (SCN1B) was the first gene identified to be associated with febrile seizures. However, very little work has been done on SCNl B gene in epilepsy patients, especially in Pakistan. The present study was
conducted to understand the comprehensive role of SCN1B gene in GEFS+ patients.
Blood samples of unrelated true representative of generalized epilepsy with febrile
seizures plus were collected from psychiatry and preventive pediatrics departments of
various hospitals of Lahore. DNA was extracted and amplified with specially designed
primers and sequencing of the peR products was also done. Analysis of the sequences
and SNPs/mutations was done with the help of appropriate bioinformatics softwares. In
the present study, polymorphism analysis of human SCNIB gene isolated from healthy
and diseased Pakistani individuals (suffering from neurological disorder, GEFs+) have
been investigated for genetic association. Novel mutation IVS2-1 G> T in splice acceptor
site of exon 3 have also been identified from Pakistani GEFS+ patients. This mutation
was absent in the healthy (control) sample. This is first report of gene characterization
and polymorphism of Human SCNI B gene from Pakistan. Likewise, in the last 15 years,
several mutations in the genes have been identified which were associated with GEFs+.
In addition to detecting new mutations and identifying new genes, further studies are
required to elucidate the particular role of furtive mutations, genetic milieu, environment,
or random events on the individual's phenotype. This study has opened a new avenue in
medical sciences in Pakistan, which will help the scientists to work on genetic disease
following the methodologies used in this study. The outcome of this study can further be
used to confirm the hypotheses through animal modeling and proteomics. The mutation
found in this study may add the information in gene databanks, which ultimately help the
scientist to develop the gene therapies for genetic diseases.
Availability: Items available for loan: UVAS Library [Call number: 1388,T] (1).
7.
Evaluation Of Pouifi As Candidate Gene To Have Influence On Milk Production In Red Sindhi And Dhani Cattle Through SNP Detection
by Waqas Tahir | Prof.Dr.Masroor Elahi Babar | Dr. Atif Hanif | Mr. Zahid Mushtaq.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2011Dissertation note: There are more than 200 million cattle in Pakistan which are a source of milk, meat, hide
and drought purpose. Due to this large population of cattle, Pakistan is the fourth largest milk
producer in the world. In spite of large number of cattle population in Pakistan, per animal milk
production is yet quite low. That's why it is not wrong to say that there is still a great potential in
the dairy sector of Pakistan to improve the overall milk production by improving the per animal
Jilk production. This can be only achieved by improving the genetics of the dairy animals in
Pakistan besides the other factors as well. Milk production is a polygenic trait. It is well
established that growth hormone (GH) released from pituitary gland also plays an essential role'
in growth, mammary gland development and lactation process. POUIFI factor is a member of
the POU family of transcription factors that regulate animal growth and development and is also
involved in the development of pituitary and regulation of hormonal expressions in animals.
Therefore, genetic variation in the POU1Fl gene and its associations with growth traits in
livestock animals could provide useful genetic markers for animal selection and breeding
through marker-assisted selection (MAS). This study was conducted to genetically characterize
!he POUIFI gene to identify the SNPs as genetic markers for future breeding and selection
~Ians. Keeping in view the importance of this gene all introns and axons were sequenced in
smples of Red sindhi and Dhanni cattle to identify the SNP and validate the potential markers
There are more than 200 million cattle in Pakistan which are a source of milk, meat, hide
and drought purpose. Due to this large population of cattle, Pakistan is the fourth largest milk
producer in the world. In spite of large number of cattle population in Pakistan, per animal milk
production is yet quite low. That's why it is not wrong to say that there is still a great potential in
the dairy sector of Pakistan to improve the overall milk production by improving the per animal
Jilk production. This can be only achieved by improving the genetics of the dairy animals in
Pakistan besides the other factors as well. Milk production is a polygenic trait. It is well
established that growth hormone (GH) released from pituitary gland also plays an essential role'
in growth, mammary gland development and lactation process. POUIFI factor is a member of
the POU family of transcription factors that regulate animal growth and development and is also
involved in the development of pituitary and regulation of hormonal expressions in animals.
Therefore, genetic variation in the POU1Fl gene and its associations with growth traits in
livestock animals could provide useful genetic markers for animal selection and breeding
through marker-assisted selection (MAS). This study was conducted to genetically characterize
!he POUIFI gene to identify the SNPs as genetic markers for future breeding and selection
~Ians. Keeping in view the importance of this gene all introns and axons were sequenced in
smples of Red sindhi and Dhanni cattle to identify the SNP and validate the potential markers
Availability: Items available for loan: UVAS Library [Call number: 1395,T] (1).
8.
Association Of Embb Gene With Ethambutol Resistance In Tuberculosis Patients
by Sana Hafeez | Dr. Atif Hanif | Prof. Dr.Masroor Elahi Babar.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Tuberculosis is a major infectious disease killing nearly two million people, mostly
in developing countries, every year. The increasing incidence of resistance of Mycobacterium tuberculosis strains to the most effective anti- TB drugs is a major factor
contributing to the current TB epidemic. Drug-resistant strains have evolved mainly due
to incomplete or improper treatment of TB patients. Resistance of M tuberculosis to anti-
TB drugs is caused by mutations in gene encoding drug targets. According to the World
Health Organization, 8 million cases of tuberculosis (TB) occur each year, resulting in 2
million deaths. TB is. transmitted by sneezing, coughing, speaking by those persons
having already TB. In affected persons mycobacterium multiplies and spread by
lymphatics to the lymph nodes, and through the bloodstream to other body sites.
Pulmonary TB is a very common type of TB. The worldwide failure of TB control
programs, combined with the HIV /acquired immunodeficiency syndrome pandemic, led
to marked increases in TB mortality. Pakistan is 1 of 22 countries listed by the World
Health Organization (WHO) as having a high incidence of tuberculosis. TB is responsible
for 5.1 percent of the total national disease burden in Pakistan. In this study sputum
samples from TB patients were collected in wide mouth, transparent containers from
local sources of Lahore. Sputa were processed with Sodium hydroxide solution to remove
contamination and inoculated directly onto two slopes 'of Lowenstein-Jensen (LJ) Medium in tubes. After inoculation samples were incubated at 37°C up to eight weeks or
till colonies appear. Colonies grown on the slopes were identified as MTB by sensitivity
and identification tests. DNA was extracted and .amplified with specially designed
primers and sequencing of the peR products was also done. Analysis of the sequences
and SNPs/mutations was done with the help of appropriate bioinformatics softwares.
DNA extraction was done in Institute of Public Health, Lahore and all remaining work
was performed in Molecular Cytogenetic and Genomics Laboratory, (IBBT), UV AS,
Lahore. Present study was related to polymorphism analysis of embB gene of MTB
isolates and its association with ethambutol resistance. From total 14 resistant samples
ten samples showed reported mutations when the query sequences were compared with
the reported reference sequence of Mycobacterium tuberculosis embB gene available on
NCBI website. None of the novel mutation was found. EMB resistant MTB strains
showed mutations in embB gene at different co dons including 306 codon and 319 codon.
Genetic analysis showed that eight samples possessed mutation at 306 codon and two had
at 319 codon. WIllie four samples did not show any mutation or alteration in embB gene
region. Patients were screened for alternations in codon 306 of the embB gene as
mutation in this codon are reported to confer resistance to ethambutol.
Availability: Items available for loan: UVAS Library [Call number: 1409,T] (1).
9.
Association Of Myogenic Factor 5 (Myf5) Gene Polymophism With Meat Quantity And Quality Traits In Sahiwal
by Faiza Maqbool | Prof. Dr. Masroor Ellahi Babar | Dr. Abu Saeed Hashmi | Dr. Atif Hanif.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Pakistan has a large population of farm animals, which are consuming for many purposes. Livestock is the major source of money for any country. Livestock is the major machines and factories which convert roughage (grasses and shrubs etc.) into quality food i.e. Meat and milk.
The Myf5 gene is a member of basic helix-loop-helix family of transcription factors which is involved in the regulation of myogenesis. Its main role in muscle growth, development, proliferation, muscle fibers formation and muscle functioning makes it a candidate gene for molecular marker of meat production in livestock.
Myf5 gene in cattle has been mapped to chromosome 5 and has a length of 3236 bp (Gen Bank accession no. NC-007303). It consists of three exons and two introns. Exons have the lengths of 659, 76 and 1245bp. Role of Myf5 gene in muscle development and growth makes it a candidate gene for meat production in farm animals. In this study association of myogenic factor 5 (Myf5) gene polymorphism with meat quality and quantity traits in Sahiwal cattle was checked out.
In this study blood samples were collected from Sahiwal cattle breeds and DNA was extracted from leukocytes. DNA amplification was done by PCR. Then sequencing of amplified gene was done by Genetic Sequencer.
Allele frequencies and genotype frequencies were statistically analyzed by using SNPator software. The relationship between SNP marker genotypes of myogenic factor 5 (Myf5) gene with meat quality and quantity traits was evaluated by using SNPator software. This study will be a helpful tool for marker assisted selection of beef cattle.
Availability: Items available for loan: UVAS Library [Call number: 1561,T] (1).
10.
Biomass Production Of Pasteurella Multocida By Using Biofermentor For Preparation Of Montanoid Based Vaccine
by Noreen Sarwar | Prof. Dr. Khushi Muhammad | Dr. Atif Hanif | Prof. Dr. Muhammad.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Hemorrhagic septicemia is a contagious bacterial disease of large ruminants principally in cattle and buffalo with high morbidity and mortality. The disease is endemic in nature and outbreaks are common during hot, humid and wet season. The acute and fatal nature and brief duration of the disease limit the antimicrobial therapy. In Pakistan, the disease causes heavy economic losses to dairy industry. Vaccination therefore, is an option for controlling the disease. For a quality vaccine, biomass production of P. multocida along with well developed capsule (immunogen) is necessary. The problem associated with the production of a quality vaccine is poor biomass production of P. multocida when grown in ordinary or routine media.
Present study was designed to isolate P. multocida from sick animals and its molecular characterization in the laboratory and study factors (temperature, media composition, pH incubation time and agitation or shaking) affecting its immunogen production and "in process quality control" factors (biological titer, dry mass, adjuvant and storage time) that affect antibody response. Finally, biomass production of the organism using biofermentor and monitoring of the antibody response of buffaloes to inactivated Montanide ISA-70 based P. multocida vaccine.
Each of the field isolates showed grey, viscous, mucoid, translucent and non hemolytic colonies on blood agar. There was no growth on MacConkey's agar. It was Gram negative coccobacilli or thin rods and bipolar when stained with Leishman's stain. The isolates were positive for Catalase, Oxidase, Hydrogen sulphide and Indole production along with nitrate reduction while it was negative for urease production, citrate utilization and gelatin liquefaction. The bacteria fermented glucose, sucrose, mannitol, mannose, but failed to ferment arabinose, maltose, salicin, lactose, dulcito and inositol.
Polymerase chain reaction (PCR) was performed on isolated colonies by using P. multocida specific and HS causing serotype B specific primers. P. multocida specific PCR gave product of 465 bp while HS causing serotype B specific primers amplified a product of approximately 590 bp.
Growth of the bacteria in casein yeast sucrose broth was optimized under different conditions. CSY broth showed dense growth of P. multocida during incubation for 18 hours. A temperature in between 35°C and 40°C showed its optimum growth. Poor growth was observed below 30°C and no growth was detected at 50°C and above. No growth occurred at pH 0.5 and 10.0 but best growth was obtained at pH 7.0 and 8.0. There was positive correlation between shaking in terms of rpm and growth. There was optimum growth at 500 rpm for 24 hours.
Inactivated HS Vaccine was prepared from dense growth in biofermentor on the basis of dry mass and bacterial count. The effect of biomass, adjuvant, storage of the vaccine, priming alone or with boosting on its potency was also studied along with boosting effect of montanoid ISA 70 oil based vaccine. Dry mass 1.7 mg/dose produced protective antibody titer while bacterial count 10-14/ml was sufficient to produce the protective antibody titer. Montanoid ISA 70 based vaccine provided immunity to buffalo calves better than aluminium hydroxide gel and bacterins. Boosting with oil based vaccine can help to keep the animal immunized for whole year. For better results of vaccine, it can be stored at 4oC for six months.
It is concluded that the proposed study improved quality of the vaccine and reduced volume of the vaccine dose, cost of its production and frequency of vaccination.
Availability: Items available for loan: UVAS Library [Call number: 1581,T] (1).
