1.
Antibody Response Of Buffalo To Inactivated Foot And Mouth Disease (Aphtho) Virus Vaccine
by Nadeem Murtaza | Prof. Dr. Khushi Muhammad | Prof. Dr. Masood Rabbani | Faculty of Veterinary Sciences.
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Publisher: 2005Dissertation note: Buffaloes when vaccinated against Foot and Mouth Disease (FMD) vaccine containing serotype "O" of the virus, showed detectable level of complement fixing (CF) antibodies. Buffaloes vaccinated with Aftovaxpur vaccine showed undetectable level of CF-antibodies when tested through complement fixation test using local vaccinal serotype "O" of FMD virus. However, buffaloes irrespective of age and sex when vaccinated with aluminum hydroxide adsorbed FMD vaccine containing serotype "O" of the FMD virus, showed detectable level of CF-antibodies, when tested through CFT using the local serotype "O" Of FMD virus. These antibodies disappeared fourth month post boosting. Buffalo calves immunized with oil based FMD vaccine showed high-level GMT titer (17.6) of the CF-antibodies. Rabbits immunized with the oil based FMD vaccine showed high level GMT (31.2) of the CF-antibodies. Low level of CF-antibodies might be sufficient to induce resistant to field exposure of the FMD virus serotype in the presence of blood complement. Sera of buffalo, cattle, sheep, goat and guinea pigs contained complement titer 35.2, 32.6, 19.2, 20.8, 614.4, respectively. Moreover, it was observed that complement activity remained stable when stored at -200C for 24 hours. The complement activity decreased from 1:512 to 192, 70.4, and 13.6 when stored at 40C, 250C and 370C, respectively. The complement activity was detectable when diluents containing Ca++and Mg++ ions were used.
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2.
Development Of Standard Protocols For Preparation And Evaluation Of Liver Homogenate Vaccines Against
by Sahidullah | prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad | Faculty of Veterinary Sciences.
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Publisher: 2005Dissertation note: Twelve vaccines were prepared from HPS infected liver homogenate by using two different virus concentrations (1x102 &1x103 LD50) and two virus inactivants (1%formalin and 0.001%Binary ethyleneimine) with and with out different adjuvants. These vaccines were evaluated in 13 groups of day old broilers (105 chicks) for their comparative immunogenicity and protection. At day 14 of age, groups A1, B1, C1 & D1 were vaccinated with 4 oil base vaccines (OB-HPSV) with different virus concentration and different inactivants. Similarly groups A2, B2, C2 & D2 were vaccinated with aluminized vaccines (AH-HPSV) and groups A3, B3, C3 & D3 with non adjuvant vaccines (NA-HPSV). Groups E was kept as unvaccinated control group. All the vaccinated birds were found sero-positive 7 days post vaccination (PV). IHA GMT results indicated no difference for different virus concentrations and different virus inactivants but same adjuvant. The IHA GMT recorded weekly during 0-28 days post vaccination was the highest and more consistent (52-181) for OB-HPSV followed by AH-HPSV (52-147) and then NA-HPSV (73.3-104). All the birds vaccinated with OB-HPSV resisted the virus challenge 21 days PV (100% protection). While protection percentage recorded for AH-HPSV and NA-HPSV was 87.5 % & 62.5% respectively. It was concluded that 1x102 LD50 oil base vaccines inactivated with either formalin or binary amine may be recommended for commercial use being the best in experimental trails.
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3.
Studies On In Vitro Culture Characteristics Of Adherent Baby Hamster Kidney-21 (Bhk-21) Cell Line
by Saddeq-ru-Rahman | Prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad | Faculty of Veterinary Sciences.
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Publisher: 2005Dissertation note: Baby Hamster Kidney-2 1 (BHK-2 1) cell line growth pattern, growth requirements, growth effectors, cryopreservation and its susceptibility to different animal viruses were studied to optimize the in vitro culture requirements and conditions for maintenance and long time cryopreservation in liquid nitrogen of this cell line. It was observed that BHK-2l cells multiplied fast during first 48 hours and made a complete monolayer after getting confluency with in 72 hours post incubation which was followed by a decline phase. Fetal calf serum has growth stimulating effect and 5 - 10% serum level was satisfactory for the maintenance of cell line. While harvesting the cells from a flask, Trypsin (0.25%) with neutralization by fetal calf serum (5-10%) was found better. For cell storage 10% Dimethylsulfoxide (DMSO) through gradual cooling maintained maximum recovery of viable cells during cryopreservation.
Footh and mouth disease virus (FMDV; serotype "0" and "A") were adapted to cell this cell line, while canine parvo virus (CPV), Newcastle disease virus (NDV LaSota strain), canine distemper virus (CDV), and hydropericardium syndrome virus (HPSV) could not adapted to this cell line through five blind passages in this study.
Availability: Items available for loan: UVAS Library [Call number: 0916,T] (1).
4.
Studies On The Physico-Chemical Factors Affecting In Vitro Replication Of Foot And Mouth Disease Virus (Serotype"O")
by Muhammad Taslim Ghori | Prof. Dr. Khushi Muhamamd | Prof. Dr. Masood Rabbani ( | Faculty of Veterinary Sciences.
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Publisher: 2007Dissertation note: Effect of physical (temperature, U.V light, and pH) and chemical factors such as sodium carbonate, sodium hydroxide, potassium permanganate, formaldehyde, citric acid, fin-virus, iodine and sodium hypochlorite on the replicating ability of"O" type of FMD virus on BHK cell line was determined.
The FMD virus of known TCID was exposed to 37, 57 and 77°C for 15, 30 and 45 minutes. Each of the virus's aliquot exposed to temperature was inoculated to 8 wells of the 96-well cell culture plate containing adherent monolayer of BI-IK cell line. The plates were incubated at 37 °C for 48 hours. The results showed that the temperature more than 57°C can inactivate the virus within 15 minutes. The virus was admixed in the MEM-199 maintenance media at pH 3, 5, 7, 9 and 11. The results showed that the virus was survived at pH 7 but virus was inactivated at pH 3, 5, 9 and 11. The FMD virus of the known TCID o was exposed to U.V. light (1 foot distance) for 15, 30 and 45 minutes. The results indicated that the virus tolerated UV light of 252.7nrn as it showed cytopathogenic effects (CPE).
The FMD virus of known TCID was exposed to 0.5 x, lx, and 2x concentration of each of the iodine, sodium carbonate, sodium hydroxide, formalin, finvirus, potassium permanganate, sodium hypochlorite and citric acid for 30, 60 and 90 minutes. Each of the virus's aliquot exposed to either of chemical factors was inoculated to 8 well of the 96-well culture plate containing adherent monolayer of the BI-IK cell line. The plates were incubated at 37 °C for 48 hours. Development of CPE indicated the virus inactivating ability of the chemical factor. The results showed that formalin, iodine, finvirus and sodium hypochiorite are more effective against FMD virus.
The results of the study are helpful to curtail the spread of virus, to implement the effective bio-security measures in our local conditions and in processing of animal products fit for export.
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5.
Seroprevalence Of Bovine Brucellosis In District Quetta, Balochistan
by Muhammad Shafee | Prof. Dr. Masood Rabbani | Dr. Mansur-ud-Din Ahmad | Faculty of Veterinary Sciences.
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Publisher: 2007Dissertation note: The sero-epidemiological study of bovine brucellosis was carried out to observe the incidence of brucellosis in slaughterhouse and Government and private dairy farm, (GDF, PDF) Quetta, Balochistan. The prevalence of this disease out of 780 serum samples of cattle and buffalo in slaughterhouse was recorded 3% by Rose Bengal Plate Test (RBPT) and 3.20% by indirect enzyme linked immunosorbent assay (i-ELISA), respectively. The zoonotic natures of this disease was also checked by screening 20 serum samples of slaughterhouse workers butchers and veterinarians and were found (5%) 01 positive out of 20 by RBPT but no positive case was found by i-ELISA. Similarly the disease was also checked in 200 milk samples of Government and Privatly owned Dairy Farm, Quetta.
The overall prevalence observed in the area by screening 1000 serum and milk samples of the target human, cattle and buffalo, was 4.2 % through i-ELISA.
The prevalence observed in Government Dairy Farm (GDF), Quetta was 14.8% (11 positive out of 74) while the Private Dairy Farm (PDF), exhibited 4.76% (6 positive out of 126 milk samples) by screening through i-ELISA. At GDF, Quetta, out of total of 74 cattle, no case were found positive by MRT, although 03 cases were found doubtful while i-ELISA show 11 positive cases in cattle (14.8%) while in private dairy farm 4 out of 15 cattle (26%) were found positive and 01 was considered doubtful by MRT and ELISA detected 06 cases of cattle out of 15(40%). Similarly 2 out of Ill (1.8%) buffalo were positive and 02 were doubtful by MRT but ELJSA did not detect any positive case and the prevalence of bovine brucellosis was higher in animals with reproductive disorders especially in cases of abortion.
The present study also revealed that the disease is more prevalent in cattle than buffalo both in slaughterhouse and organized dairy farm (Govt and private). In slaughterhouse 12 out of 23 cases were found positive by RBPT and 22 out of 23 were found positive by i-ELISA while in organized dairy farm all of the 17 milk samples were found positive from cattle population.
The efficacy of the i-ELISA both for milk and serum samples was found higher than other two conventional tests (MRT and RBPT), as it detected higher percentage of brucellosis cases both in serum and milk samples in comparison to other two tests.
The results of this study have revealed an alarming situation of bovine brucellosis in our dairy animals, which needs an emergent response from policy makers, as the disease is a potential threat to the human and animal health.
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6.
Effects Of Different Disinfectatnts On Pathogens In Poultry
by Asif Abbas Malik | Prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad | Faculty of Veterinary Sciences.
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Publisher: 2006Dissertation note: Poultry sector is the second largest industry after textile in Pakistan. It is threatened by various diseases i.e; Newcastle disease (ND), Avian Influenza (Al), Colibacillosis, Hydro-pericardium syndrome (HPS) and Infectious bursal disease (IBD, Gumboro). The efficacy of various available disinfectants (Hygen 275 — 2000 H, Virkon S and Aldekol) was tested at 2x, lx and Y2 x dilution against Staphylococcus aureus, Escheria Coil, Newcastle Disease virus and Avian Influenza virus. Each dilution of all the disinfectant was divided into 4 aliquots i.e; a, b, c and d (each of the
aliquot, for each pathogen). Each aliquot were mixed with equal volume of either of the pathogen. The mixture of the disinfectant and the pathogen was incubated at 37°C for a period of 15, 30 and 45 minutes of interaction. The contents were collected aseptically and processed to evaluate the effectiveness of the disinfectants. Disinfectant A (Hygen 275-2000H) showed good bactericidal as well as virucidal activity at 1% dilution. Disinfectant B (Virkon S) was able to kill all the bacteria and viruses even at 0.25 % dilution. While, disinfectant C (Aldekol) effectively killed the bacteria and viruses at 0.5 % and I % dilutions. Results of the study will help the farmers to adopt effective biosecurity measures to minimize the challenges at farm level.
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7.
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.
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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.
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8.
Prevalence Of Multiple Drug Resistant (Mdr) Bacteria In Intestinal Infections Of Dogs
by Iffat Habib | Dr. Aftab Ahmad Anjum | Prof. Dr. Masood Rabbani.
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Publisher: 2011Dissertation note: Antimicrobial resistance is a complex problem involving various bacterial species, resistance mechanisms, transfer mechanisms and reservoirs. Cats and dogs are the potential sources for spread of antimicrobial resistance in humans due to their close contact with them. The horizontal transfer of antimicrobial resistance genes through plasmids, integrons and transposons has been found to play an important role in the dissemination of antimicrobial resistance genes. Canine antimicrobial resistant genes had been identified in bacteria isolated from human clinical infections suggesting the spread of resistance mechanisms from canine to human bacteria.
The present study has been designed to study the prevalence of multiple drug resistant strains causing enteritis in dogs. 100 Samples were collected from different Pet clinics in and around of Lahore city. These samples were cultured for identification of MDR bacteria. Antibiotic resistance profile was studied by the standard Disk diffusion method (Kirby-Bauer Method) for commonly used antibiotics. These MDR bacteria were isolated and identified as per standard protocols described in the Bergey's Manual of Determinative Bacteriology. Different combinations of antibiotics were also evaluated for in-vitro antibiotic sensitivity for an effective treatment of these cases so that the load of MDR bacteria could be reduced.
From the collected samples E. coli, Salmonella enterica, Proteus vulgaris, Citrobacter diversus and Psedomonas spp. were identified. Among all of these E.coli was most prevalent followed by Salmonella enterica, Citrobacter diversus, Proteus vulgaris and Psedomonas spp. Out of 127 E.coli isolates 52 40.94%) were declared as MDR-Bacteria following 50 Salmonella enterica isolates 17 (34.00%), 17 Citrobacter diversus 6 (35.29), 12 Proteus vulgaris isolates 06 (50%). It was concluded that MDR isolates were most sensitive to antibiotic combination (Amoxicillin + Clavulanic Acid), followed by (Oxytetracyclin + Tylosin), (Gentamycin + Ceftriaxone), and (Penicillin + Streptomycin). Out of 52 MDR E.coli isolates 23 (44.23%) were found to be invasive. Recommendations are made on prudent use of antimicrobial drugs in dogs, as well as on the need to develop science-based infection control programs in veterinary hospitals.
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9.
Uterine Microbial Flora Of Sahiwal Cattle During Oestrus And Its Relayionship With Pregnancy Rate
by Habib- Ur- Rehman | Prof. Dr. Masood Rabbani | Dr. Ali Ahmad Sheikh | Prof. Dr. Nasim.
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Publisher: 2011Dissertation note: In the present study uterine microbial flora of Sahiwal cattle during oestrus and its relationship with pregnancy rate was determined. According to the results a total of 11 bacterial species were isolated from 50 uterine samples of estrus Sahiwal cattle, maintained at Livestock Production Research Institute (LPRI), Bahardur Nagar, district Okara, Punjab province, Pakistan. The isolates include E. coli, Micrococcus spp., Bacillus spp., Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus spp., Pseudomonas spp., Citrobacter diversus, Salmonella spp., Proteus spp. and Corynebacterium spp. Tabulation of results showed that prevalence of these isolates was different among pregnant and non-pregnant animals. Moreover, E .coli, Micrococcus spp., Bacillus spp., Staphylococcus aureus and Citrobacter diversus are found to be thriving in uterus as normal microbial flora, whereas, Streptococcus spp. isolate as abnormal microbial flora appearing to be having some role in decreasing pregnancy rate. While, Pseudomonas spp., Corynebacterium spp. Staphylococcus epidermidis, Salmonella spp., and Proteus spp. Isolates could not be differentiable as normal and abnormal uterine microbial flora due to insignificant available data. Furthermore, complete blood counts of 50 blood samples of these same animals indicated that those animals harboring isolates like Streptococcus spp., Pseudomonas spp., and Corynebacterium spp. in their uterus, had more likelihood of abnormally increased value of Mean Corpuscular Volume (MCV) than to presence of any other bacteria. But due to lower data of Pseudomonas spp., and Corynebacterium spp isolated from total samples, only Streptococcus spp. seemed to be ranked as abnormal in Pakistani Sahiwal cattle cows. Interestingly all those animals from where Corynebacterium spp. was isolated, were showing increased values both of MCV and HCT (Hematocrit) which is indicative of their pathogenic role in causing uterine infections.
On the basis of this study it can be modestly concluded that uterine microbial flora identification may serve as a better tool in assessing and foretelling the reproductive health status of the breeding animals. After necessary assessment, presence of any harmful microbial flora or pathogen can be effectively treated through either selecting an appropriate antibiotic by using culture sensitivity testing or by using any suitable bactericidal agent thereby help in boosting conception and pregnancy rates.
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10.
Uterine Microbial Frlora Of Nili Ravi Buffalo During Estrus And Its Relationship With Pregnancy Rate
by Sohail Raza | Prof. Dr. Masood Rabbani | Dr. Ali Ahmad | Dr. Muhammad Iqbal.
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Publisher: 2011Dissertation note: The low conception rate has been reported as one of the major cause of poor productivity of livestock. Beside other reasons, presence of different types of microflora inside the uterus of breeding animals, play a key role in the failure of pregnancy. All these microbes results in the infection of uterus ultimately affecting drastically the animal's conception rate. To study the impact of microbial flora on conception rate, 50 Nili Ravi buffalos were selected from Buffalo Research Institute, Pattoki. The breeding animals in heat just before artificial insemination were used to collect bacterial samples with the help of especially prepared and sterilized AI rod with some accessories. The samples were processed for the identification of bacterial microflora by doing number of conventional tests for final characterization. In this study seven different bacterial isolates were identified from all the samples. These include: Escherichia coli, S. aureus, S. epidermidis, Citrobacter species, Proteus species, Lactbacillus species, and Micrococcus species. After elapse of proper period of time the pregnancy statuses of all these buffaloes were determined and correlated with the presence or absence of isolated microbes. The results indicated that Escherichia coli and Staphylococcus aureus isolates were the most prominent bacteria in all the samples collected from pregnant, non pregnant and aborted animals. These two isolates could be designated as normal uterine microbial flora of Nili-Ravi buffaloes because of their presence during all the physical and pathological conditions. Proteus species and Micrococcus species were mostly isolated in pregnant animals. Statistical analysis also confirmed the above statement. Previous reports corroborate the present study and confirm that these bacteria are ranked as normal uterine microbial flora of bovines. So the previous study and present results confirm that both are the normal uterine microbial flora of pregnant Nili-Ravi buffaloes In the present study the prevalence of the Citrobacter spp. only in the aborted animals is supported by the previous studies which show that Citrobacter spp. Is only present in the diseased animals and it also cause the sporadic abortion. Statistical analysis of the data also proved the significance of Citrobacter spp. in aborted animals. So this concludes that Citrobacter spp. are the abnormal uterine microbial flora of Nili-Ravi buffaloes in Pakistan which leads to abortion. The present study has been able us to find the normal and the abnormal uterine microbial flora of Nili-Ravi buffaloes. This information will help to understand the infection process in breeding buffaloes and through corrective actions may decrease the infection rate / abortion rate in Nili-Ravi buffaloes.
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11.
Passive Immunization Of Infectious Bursal Disease Virus Infected Birds Using Chemically Purified Immune Yolk
by Ammara Akram | Prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad.
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drama
Publisher: 2011Dissertation note: Infectious bursal disease (IBD) is a major killer disease of poultry. It is also known as
Gumboro disease where the disease was reported first time. It is double stranded RNA
virus belongs to the family birna viridae. This disease is quite endemic in Pakistan
which has huge impact on poultry industry. Besides vaccination if immune yolk is
properly harvested and purified it can be used for treating of IBDV infected birds.
Therefore this work has been outlined to study the effectiveness of immune yolk in
experimentally produced IBDV infected birds. Refinement of yolk IgY from egg yolk
of immunized hens. Suitability in using hyperimmune egg yolk in IBD infected bird
in field conditions. In order to get hyper immunized egg yolk 20 commercial layers
were raised in poultry shed of the university. They were supplied with fresh water and
feed ad libitum with proper hygienic condition. The birds were vaccinated with oil
based killed Gumboro vaccine twice at 15 days interval at the age of 26 weeks to get
immune yolk. Eggs were collected at two weeks interval till two and half months after
boosting. Immune yolk was purified by chemical means. Antibody against IBD in egg
yolk and semi purified egg yolk IgY was measured by indirect ELISA kit method.
The eggs which were collected 15 days interval after boosting had the highest
antibody titre which decline with the passage oftime and lowest was recorded 75 days
after boosting. Similar pattern of results were also observed in semi purified egg yolk.
However significant antibody titre was lost during purification process. 50
commercial chicks of 15 days old were purchased and they were reared in poultry
shed in the university up to 36 days. They were splitted in eight groups and two
experiments were carried out side by side. In experimental chicks the birds were
challenged with the Gumboro infected bursal homogenates which were confirmed by agar gel diffusion tests. In first experiment the birds were challenged at the day of 30 days and they were provided with passive therapy of immune yolk and semi purified
IgY after 3 days of challenge. In the second experiment the birds were challenged and
passive immuno therapy was provided 24 hours interval of challenge and concurrently. The birds which received semi purified immune yolk and antibody titre having more or less 4000 they showed 20% mortality in the each group.
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12.
Comparison Of Diagnostic Approaches For The Detection Of Bovine Viral Diarrhea Persistency In Dairy Herds
by Arfan Ahmad | Prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad.
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Publisher: 2012Dissertation note: Bovine viral diarrhea is one of the most important diseases of cattle which are causing
continuous economic losses to the cattle industry primarily due to decreased reproductive
I performance. Without doubt, direct contact between BVDV persistently infected, and susceptible animals is the most important transmission route of virus. All control programs which are in use in many countries of the world, mainly depend upon the detection of PI animals, eliminating them and preventing their return into the herds. Therefore, in this study diagnostic suitability of ear notch biopsies and serum samples were compared for the detection of PI animals, as well as proficiency of various diagnostic approaches like VI, AC-ELISA, IHC and real time RT-PCR were evaluated using ear notch biopsies. A total of 468 samples were collected from 12 participating dairy cattle farms located at Prince Edward Island, Canada. The samples were divided into two groups on the basis of age, A " 6 months), and B (> 6 months).
PI calves remain immunotolerant to the infecting strain but if exposed to a heterogonous strain postnatally, they may develop low level of antibody. Accordingly, serum neutralization was applied for initial screening of samples for further testing. The samples of animals of group B, having SNT (:S 1 :64) were selected, while all samples of younger aged group A were processed without considering the serum neutralizing titres, because unlike older animals, P.1. animals below 6 months of age can have passive colostral antibodies in the course of persistency. Diagnostic suitability of ear notch biopsy and serum sample for confirmation of BVDV
A significant discrepancy was observed between ear notch biopsies (51198 positive) and
serum samples (71198 positive) during first round of testing by real time RT-PCR. However, on
follow up testing, 30 days post first round of testing, a complete agreement between ear notch
biopsies and serum samples was observed. On second round of testing, a total of 4 animals out of
197 (one positive animals died before re-sampling) were confirmed with PI, using both ear notch
biopsies and serum samples. The decrease in the positivity using RT-PCR on serum samples in
the second round of testing reflected the presence of 2 transiently infected animals. Ear notch
biopsy (EN) testing did not detect any transiently infected animal indicating the lack of
delectability of the virus in EN during transient infection under conditions of this study. After
follow up testing, 2 animals in each of group A and B were identified as PI. These findings have
led us to conclude, that either serum or ear notch biopsy can be used for the detection of
persistent infection. Of 468 collected and 197 tested samples, an overall 0.85% and 2.03%
prevalence of PI animals with BVDV was observed respectively. A complete agreement (P value=l) was observed when three diagnostic approaches (Real time RT- PCR, AC-ELISA, and IHC) were compared with standard of VI. A total of 197 ear notch biopsies (145 of group A and 52 of group B) were tested by the four diagnostic tests, four animals (2 from group A and 2 from group B) were found positive by all the tests applied. A complete agreement was observed between the first and the second round of testing. All four assays were found specific but real time RT-PCR was found to be more sensitive. Both, VI and IHC were found labour intensive, as diagnosis may take more than one week to be made. Further PI calves remain immunotolerant tothe infecting strain but if exposed to a heterogonous strain postnatally, they may develop low leved
ofantibody. Accordingly, serum neutralization was applied for initial screening of samples for further testing. The samples of animals of group B, having SNT (:S 1 :64) were selected, while all samples of younger aged group A were processed without considering the serum neutralizing titres, because
unlike older animals, P.1. animals below 6 months of age can have passive colostral antibodies in the course of persistency. Diagnostic suitability of ear notch biopsy and serum sample for confirmation of BVDV persistent animals were evaluated by real time RT-PCR. TaqMan probes and primers specific for BVDVI and BVDV2 were used. They were found specific and able to detect 10·s and 10-4 TCID50 units ofBVDVI and BVDV2, respectively.
Availability: Items available for loan: UVAS Library [Call number: 1407,T] (1).
13.
Characterization Of Antimicrobial Resistance In Escherichia Coli Recovered From Retail Chicken In Lahore
by Fayyaz Yasin | Dr. Ali AHmad Sheikh | Prof. Dr. Masood Rabbani.
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Publisher: 2012Dissertation note: Escherichia coli is normal inhabitant of lower intestinal tract of all warm blooded animals. The background of drug resistant studies on E. coli represent that extensive and irrational use of antimicrobials in human and veterinary sector for treatment, prophylaxis and feed additive made this organism resistant to commonly used antimicrobials.
It is assumed that E. coli have the ability to transfer resistant genes via bacterial conjugation, transduction and transformation. As a result pathogenic organisms are also becoming resistant to commonly used antibiotics. And there is a need to check the extent of resistance to ensure the efficacy of antimicrobials used in public health.
The purpose of current study was to estimate the prevalence of antimicrobial resistant E. coli in retailed chicken meat samples of Lahore city. In current study hundred chicken meat samples were collected from local market in various areas of Lahore.These samples were processed for isolation of generic E. coli. Initial confirmation of generic E. coliwas made using standard culturing and biochemical reactions. The prevalence rate of E. coli was found 85%. The antimicrobial resistance against in the E. coli isolates was determined by disk diffusion method. All the E. coli isolates were found resistant to at least seven antimicrobials and 34 different antimicrobial patterns were found.
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14.
In Process Quality Control Factors Affecting The Potency Of Indigenous Mycoplasma Gallisepticum (Mg)
by Muhammad Asim Raza | Dr. Arfan Ahmad | Prof. Dr. Masood Rabbani.
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Publisher: 2012Dissertation note: Mycoplasma gallisepticum (MG) is cause of chronic respiratory disease (CRD) and is responsible for significant economic losses to poultry industry. In Pakistan, imported MG bacterin fails to induce immunoprophylaxis that could be due to subtle antigenic variation in the immunogen of the vaccine. Therefore present study was conducted to optimize inactivations( phenol, formalin and binary ethylenimine) concentration and exposure time to inactivate MG and their effect on potency of MG bacterin ( prepared from local isolates) along with different bacterial biomasses (0.5%, 1% and 1.5% PCV) and adjuvants (montanide oil ,gel and water ). It was observed that the MG bacterin containing 1.5% level of immunogen/biomass induced significantly higher anti-MG-ELISA antibody titer (p < 0.05) as compared to other bacterins containing lower concentrations of the immunogen.The formaldehyde inactivated the pathogen within shortest possible time and showed undetectable effect on its potency. The antibody response was significantly higher (p<0.05) as compared to that of bacterins prepared from the pathogens inactivated by either phenol or BEI. . Montanide ISA70 containing MG bacterin induced significantly higher anti-MG-ELISA antibody titer (p<0.05) in broilers than the other bacterins containing either water or aluminum hydroxide gel. It is concluded that formaldehyde inactivated oil based vaccine containing one percent immunogen (0ne percent PCV) induce antibody response in broilers that is comparable with the imported vaccine.
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15.
Molecular And Serological Characterization Of Avian Influenza Viruses In Domestic And Wild Birds
by Mobeen Sarwar | Prof. Dr. Khushi Muhammad | Prof. Dr. Masood Rabbani.
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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).
16.
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.
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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).
17.
Characterization Of Mycoplasma Gallisepticum Isolates And Their Use In The Production Of Indigenous
by Mushtaq Ahmad | Prof. Dr. Masood Rabbani | Prof. Dr | Prof. Dr. Tahir Yaqub.
Material type: ; Format:
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drama
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1564,T] (1).
18.
Transovarian Transmission And Molecular Characterization Of Hydropericardium Syndroe Virus In Experimentally Infected Poultry Birds
by Rabia Tahir | Prof. Dr. Khushi Muhammad | Prof. Dr. Masood Rabbani.
Material type: ; Format:
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drama
Publisher: 2013Dissertation note: Poultry is one of the important and growing industries in Pakistan. It has now become the tremendously growing sub-sector of livestock. But the major constraint in growth of this industry is infectious diseases of poultry which leads to high mortality and morbidity rate resulting in heavy economic losses. The present study was conducted on one of the infectious poultry diseases named Hydropericardium syndrome which commonly affects broilers between 3-5 weeks of age. The causative agent is highly infectious virus belonging to Avian Adenovirus serotype 4. These are non-enveloped DNA viruses. It is an acute, infectious disease characterized by high mortality and excess pericardial fluid and multifocal hepatic necrosis. The incubation period ranges from 2-5 days followed by inoculation with liver homogenate or purified virus. This disease is characterized by the accumulation of straw coloured jelly like fluid in the pericardial sac, discoloured and inflamed liver with basophilic intranuclear inclusion bodies, congested kidneys and mortality up to 70%.
The present research was planned to prove that hydropericardium syndrome virus besides horizontal transmission is also transmitted by vertical transmission through transovarian route. For this purpose Liver sample was used as source of this virus. These liver samples were processed for further propagation of virus in live birds. Moreover, Virus neutralization test was also conducted for the confirmation of virus. To prove the transovarian transmission of this disease liver homogenate of infected birds was injected in 22 wk old breeder. Eggs were collected at 7-14, 15-21 and 22-28 days post infection. The day old hatched chicks were slaughtered to obtain liver and spleen sample for confirmation of virus through PCR. For confirmation, DNA was extracted using KIT method followed by polymerase chain reaction and amplified genomic material was visualized through gel doc system.
After validation of PCR 90 samples of liver and spleen were processed for DNA extraction followed by PCR. None of the samples of extracted DNA processed for each tissue from chicks produced a visible band of DNA in agarose gel after ethidium bromide staining. The possible reason for these negative results may be that viral load was below detectable limit or the presence of high titre of neutralizing antibodies. The implications of these findings are that vertical transmission via transovarian route does occur in poultry birds but the exact mechanism and establishment of latent infections further need to be investigated.
Availability: Items available for loan: UVAS Library [Call number: 1585,T] (1).
19.
Antibacterial Activity Of Herbal Extracts Against Multi-Drug Resistant Escherichia Coli Recovered Form Retail
by Arfat | Dr. Ali Ahmad Sheikh | Prof. Dr. Masood Rabbani.
Material type: ; Format:
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drama
Publisher: 2013Dissertation note: Escherichia coli is normally present in lower part of intestinal tract of all warm blooded animals. The background of drug resistant studies on E. coli represents that extensive and irrational use of antimicrobials in human and veterinary sector for treatment, prophylaxis and feed additive made this organism resistant to commonly used antimicrobials. Retail chicken meat is one of the major sources of spread of MDR E. coli infections in humans.
The purpose of present study was to evaluate the prevalence of multi drug resistant E. coli recovered from retail chicken meat samples collected from various areas of Lahore city. Also study the effect of antibacterial activity of selected herbal extracts against isolated MDR E. coli. In current study 100 E. coli isolates were processed for isolation of generic E. coli. Identification of generic E. coli was made using standard culturing, biochemical reactions and confirmed through PCR. The recovery rate of E. coli was found 80% and multidrug resistant pattern in the E. coli isolates was determined using disk diffusion method. A total 73.86% of E. coli isolates were found resistant with at least three antimicrobials related to different groups.
Current study revealed the effectiveness of herbal extracts against MDR E. coli. Clove, cinnamon and mint have good antibacterial activity as compared to coriander, kalonji and garlic. Hence, these herbal extracts can be used as promising alternatives of antimicrobials against multiple drug resistant E. coli species.
Availability: Items available for loan: UVAS Library [Call number: 1606,T] (1).
20.
Prevalence Of Salmonella And Campylobacter Contamination In Poultry Eggs
by Hassaan Bin Aslam | Dr. Aftab Ahmad Anjum | Prof. Dr. Masood Rabbani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2013Dissertation note: Provision of adequate food to their inhabitants and assure an atmosphere free from hunger and malnutrition is the responsibility of a civilized government. The food security objective becomes more important when 15-20% of the world population is not getting sufficient food to meet minimum nutritional requirements for a healthy and productive life. Proteins play an important role in the formation of balanced human diet. There are mainly two sources of proteins i.e. animals and plants. Commercial egg production is an important economic enterprise offering more rapid and efficient return than many other livestock production operations. In 1999-2000, 13.9 million commercial layers in Pakistan produced 3,261 million eggs contributing 27% eggs to the total egg production of 8677 million eggs.
The incidence of food borne diseases is increasing globally. Many cases of food borne illness occur as a result of improper food handling and preparation by consumers in their own kitchens. Some of the most compelling evidences have come from the international data on Salmonella and Campylobacter species infections. Food-producing animals (e.g., cattle, chickens and turkeys) are the major reservoirs for many of these organisms. A total of 500 raw chicken eggs were bought from different retail outlets in Lahore city for determining the prevalence of Salmonella and Campylobacter inside and on the egg shell and enumeration of their load. Samples were graded as clean, trace, dirty and cracked. All samples were processed inside the safety cabinet. Eggs were broken from narrow end and contents were drained in the petri plate. Egg shell and egg shell membrane was processed by shell crush method in a tube in the presence of phosphate buffer saline. Sample from egg yolk and albumin was taken by direct aspiration while egg shell rinse was taken as a sample for Salmonella and Campylobacter. Isolation is done by enrichment method for this purpose selenite broth and buffered peptone water is used for Salmonella and campylobacter, respectively. The enriched sample was then plated on the Brilliant Green Plate selective for Salmonella and Campy Cefex Agar plate that is selective for Campylobacter. Thirteen samples out of 500 hundred samples were positive for presence Salmonella with over all prevalence of 2.6%. Highest percent prevalence was found in cracked eggs where it is 33.3% followed by dirty and clean eggs (0.9%) and trace eggs with zero prevalence. Colony forming units of Salmonella on the shell of one positive sample is 4.2x102 while CFUs in egg yolk of cracked egg was 7.0x102. Regarding Campylobacter five eggs out of 500 eggs were positive with overall prevalence of 1%. The highest prevalence was found in cracked eggs where it was 16.6% and dirty eggs having prevalence of 12.5%. Both clean and trace eggs were having zero prevalence. CFUs for Campylobacter were too low to count while majority of samples were observed negative for viable CFUs for Campylobacter.
Availability: Items available for loan: UVAS Library [Call number: 1614,T] (1).
21.
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).
22.
Molecular Identification Of Soil Borne Bacillus Anthracis From Districts Lahore And Sheikhupura
by Tariq Jamil | Prof. Dr. Masood Rabbani | Dr. Muhammad Zubair Shabbir | Prof. Dr.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2013Dissertation note: Background:
Anthrax is a bacterial zoonotic disease caused by Bacillus anthracis. Accurate assays for etiological identification are necessary to ensure proper veterinary and medical health facilities against such diseases. Real-time PCR is a powerful technique to identify this organism based on the presence of two unique plasmids (pXO1 and pXO2) and is highly preferable technique over conventional detection assays in clinical and environmental samples both.
Methodology:
Real Time-PCR technique was used to identify Bacillus anthracis bacteria in the soils of districts Lahore and Sheikhupura. Soil samples were collected from each village of both districts and processed for genome extraction using commercial soil DNA extraction kit. Following genome extraction, the samples were run further for real-time PCR analysis. Positive controls, primers and probes were provided by the Penn state University. SPSS software and pearson's chi square distribution test were used for statistical analysis.
Findings and Suggestions:
Real-time PCR was found as a powerful tool to detect Bacillus anthracis in environmental samples. The bacterium detected was of non-virulent type and showed associations with soil humidity and land use. Further studies may include study of the bacterium with respect to soil-chemistry and sero-prevalence among positive areas of the two districts. Strain characterization is also recommended. The present results may also help in ecological niche modelling by using spatial mapping techniques. Such studies will help in a better understanding of soil as a reservoir for zoonotic organisms and surveillance of the diseases.
Availability: Items available for loan: UVAS Library [Call number: 1817,T] (1).
23.
Comparative Analysis Of Respiratiory Microbiota From Clinically Healthy And Deseased Broiler Breeders
by Husnain Ahmed | Prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad.
Material type: ; Format:
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drama
Publisher: 2014Dissertation note: The pulmonary system of birds is a considered a reservoir of innumerable bacterial pathogens including those which are subject to public health significance. This scenario makes respiratory tract of birds prone to many bacterial infections as well. There are many respiratory outbreaks in poultry that causes huge economic losses and a number of bacterial pathogens either acting as primary or secondary pathogen can be held responsible for these losses. A very small fraction of (<1%) of bacterial species are culturable, limited as well as highly variable and time consuming conventional microbiological procedures have typically excluded the normal flora present in the respiratory tract or have restricted the analysis to potential pulmonary pathogens. Due to unculturable nature of many bacterial species there is a very little room left for discovering or determining novel organisms or pathogens and their association with clinical outcomes through conventional microbiological procedures. With the advancement of technology metagenomic analysis of a given sample has emerged as a major culture independent technique for identification of many pathogens, by reading the hypervariable region of the 16S rRNA gene, culture-independent technique such as 454-pyrosequencing, can provide species specific sequence of any bacteria in a given sample.
A total of 12 T-BAL samples from breeder birds were selected based upon the quality and quantity of the double-stranded gDNA. Using 454 bar-coded pyrosequencing, the hypervariable regions of the 16S rRNA gene corresponding to V1 – V5 (~ 1,000 bp) were sequenced. Of the high-quality reads obtained (296,811) using the MOTHUR platform, the sequences were processed for sequence alignment with the 16S RDP database via BLASTn, and subsequent taxonomic analysis through MEGAN programs using a homology-based method to bin sequence reads. The results of study indicate that birds harbor a diverse microbial community including number of phyla, families, genera and characterized bacterial species. The bacterial communities were relatively conserved at the phylum level; however, at lower taxonomic levels, differences were observed in the phylotypes and abundance between the clinically diseased and healthy birds. As indicated by the rarefaction plot and the Shannon-Wiener and Simpson-Reciprocal diversity indices, the biodiversity and richness in the taxonomic content was higher in the clinically healthy birds compared with the diseased birds. Regardless of the bird health status a number of new species were identified. A number of these bacterial species have been found to be associated with infectious diseases in humans and other species, although the clinical importance of these bacteria as part of the respiratory microbiome of birds has not been established.
As the nature of bacterial species is to constantly act with one another and, potentially, with the birds themselves provides an interesting avenue for continued research. There is a need to conduct further clinico-pathological studies to establish the link between causes versus effects.
Availability: Items available for loan: UVAS Library [Call number: 1826,T] (1).
24.
Study On Micobiological Quality Of Water Supplied To Poultry Birds From Tube Wells Water Pumps Drilled Up To Varying Bore
by Sidra Moqddes | Prof. Dr. Masood rabbani | Dr. Jawad Nazir | DR.Yasin tipu.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1922,T] (1).
25.
Role Of Water Chemistryand Stabilizers On The Infectivity Of Virus In Live Neqcastle Disease Virus Vaccine
by Sahrish tariq | Dr Jawad Nazir | Prof. Dr. Masood Rabbani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1927,T] (1).
26.
Correlation Of Deifferent Managment Systems And Facilities Of Retail Milk Shops With That Of Microbial Load In Raw And Pasteurized Milk
by Faria kanwal | Prof. Dr. Masood rabbani | Dr. Ali ahmad sheikh | Dr.Wasim.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1953,T] (1).
27.
Detection Of Gram Negative Foodborne Pathogen From Retail Quail Meat Through Potimized Multplex Pcr
by Amna kanwal | Dr.Aki ahmad sheikh | Dr.Tanveer | Prof, Dr. Masood rabbani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1957,T] (1).
28.
Evaluation Of Drinking Water Quality At Zoo And Various Public Places In Disteict Lahore
by Maimona jibreel | Dr. Arfan ahmad | Dr.Hassan | Prof. Dr. Masood rabbani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1959,T] (1).
29.
Comparative Study On Prevalence And Antimicrobial Susceptibility Pattern Of Extendedspectrum B- Lactamase
by Karam rasool | Prof. Dr. Masood rabbani | Dr. Ali ahmad sheikh | Dr. Muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1992,T] (1).
30.
Impact Analysis Of Quality Control Practices In Selected Microbiology Based Veterinary Diagnostic Labs Operational In District Lahore
by Faiza Marrium (2009-VA-239) | Prof. Dr. Masood Rabbani | Dr. Ali Ahmed Sheikh | Dr. Yasin Tipu.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Veterinary diagnostic labs are the powerful ally for the diagnosis, prevention and monitoring of animal diseases in any country. Labs are ordered for test for many reasons. Errors could be present in these tests. So to avoid detrimental effects of these errors a quality control system is required. This system is maintained by following the standards given by recognized international organizations like ISO, OIE, WHO, FAO and CDC etc. These authorities make the standards which should be followed by the VDLs to improve their quality of tests and management to give precise and accurate results which will help them in being well reputed lab which give internationally accepted quality of results.
In this study 4 private and 4 public sectors Veterinary diagnostic Labs was selected and it was assured to labs that their information will remain confidential as data was used only for the research purposes. Labs were identified by the codes given to them for study purpose. Information required for this study was gained through a questionnaire. Information regarding identity of lab, contact numbers, location and type of testing/diagnosis was gathered. Information about the parameters of quality control like building design, power backup, sections of lab, operating equipments, development of log books, availability of certificate of analysis for chemicals, availability of material safety data sheet was gathered. Quality assurance issues were also addressed by gathering information about internal quality assurance program, proficiency testing etc. Then this data was analyzed by using SPSS to interpret the results.
The data was analyzed statistically through frequency distribution by using Statistical Package for Social Sciences (SPSS) version 18.0 for development of Graphs and Tables.
Summary
53
requirements about personnel and equipments were 80% and 87.5% while minimum values were 40% and 25% respectively. Maximum value about quality control measures and waste management were 89.47% and 70% and minimum percentages were 36.40% and 40% respectively. Results have shown that 100% requirements of environmental monitoring and customer care were fulfilled by some labs while some labs only fulfill 20% of these parameters.
Conclusion
This study shows that in Lahore district veterinary diagnostic labs are not giving proper attention to quality of their system and there is no significant difference between setups of private and public sector laboratories. Availability: Items available for loan: UVAS Library [Call number: 2277-T] (1).
31.
Prevalence And Risk Analysis Of Coxiella Burnetii In Soil Of Sheikhupura And Attock Districts Of Punjab
by Sidra Akram (2009-VA-246) | Dr. Muhammad Zubair Shabbir | Prof. Dr. Masood Rabbani | Dr. Waseem Shahzad.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Background:
Besides nutrients and minerals, soil is aggregate of number of pathogens. Many of them are of
zoonotic importance and have significant threat to public health. Of these is Coxiella burnetii
that has been reported from other countries including the neighboring to Pakistan. Its occurrence
in soil, clinical significance and importance to human and animal health has been reported;
nevertheless nothing is known of C. burnetii in Pakistan particularly in rural setup where human
and animals are in close proximity to each other as well as the fact that how different risk factors
can be implicated in its spread and survival in the soil. PCR helps to identify the organism on the
basis of its genome and it is highly preferable over other conventional detection assays.
Methodology:
PCR technique was used to identify C. burnetii in the soils of Sheikhupura and Attock
districts.Soil samples were collected from each village of the both districts and processed for
genome extraction using commercial soil DNA extraction kit. Following genome extraction, the
samples were run further for PCR analysis followed by standard gel electrophoresis technique.
Later the pathogens prevalence has mapped in relation to roads, canals, rivers and drains for both
districts.
Summary
47
Outcome:
Contribute to the understanding about previously unrevealed prevalence of Coxiella burnetii in
soil of district Sheikhupura and Attock together with risk factor analysis implicating possible
health significance as well as survival in the soil. The distribution among two districts showed a
close association of gene IS1111 positivity and the land use. Positive samples were mostly found
along the roads and water bodies (canals, drains, river etc.). Availability: Items available for loan: UVAS Library [Call number: 2314-T] (1).
32.
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).
33.
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).
34.
Effect Of Lactobacillus Bulgaricus On Cutaneous Wound Healing In Mice
by Sonia Akram (2009-VA-160) | Prof. Dr. Masood Rabbani | Prof. Dr. Irshad Hussain | Dr. Hafsa Zaneb.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Integrity loss of major part of the skin as a result of injury, surgery, or sickness can lead to a major disability or even death. The major complication which comes in healing process is infection. Besides reports of side effects, a diversity of chemical agents is in use to facilitate wound healing process. Use of probiotics for the purpose is an approach of altering the microbial environment of wound where beneficial bacteria compete with pathogenic bacteria. Lactobacillus bulgaricus extracellular polymeric substance (EPS) contains high levels of phosphate groups which help activate macrophages and lymphocytes.
Lactobacillus bulgaricus was isolated from sources like: homemade curd, yogurt, and commercial probiotic product (Protexin, UK) as per WHO guideline. For bacterial identification, the isolates were further characterized using biochemical tests. Male mice (n= 60) were kept and divided into four groups having 15 mice in each group (A to D). Experimental treatments using suspension of Lactobacillus bulgaricus, pure honey and sterile normal saline were applied to surgically created wounds on skin of mice belonging to three groups (A, B, and C), whereas, one group (D) was left untreated. After application of treatments, tissue samples were collected from these groups, processed and examined for histopathology. Increase in number of macrophages, neutrophils and fibroblasts at wound site of mice of all groups, was recorded during histopathological examination of H & E stained slides. Results were tabulated using one-way ANOVA.
Experimental groups A and B (L. bulgaricus and honey) showed significant increase in the cells as compared to control and negative control groups (C and D). L. bulgaricus treated group
CHAPTER 6
SUMMARY
Summary
61
showed marked healing of cutaneous wound than control and negative control groups. It was observed that in L. bulgaricus treated group there was marked increase in healing process as inflammation phase of wound passed quickly. Whereas, when healing efficacy of Lactobacillus bulgaricus and honey treated groups was compared, honey was efficacy of cutaneous wound healing was significantly more than Lactobacillus bulgaricus in terms of wound contraction and subsiding inflammation. It is concluded that holistically use of honey for wound healing in mice is more efficacious therapy than use of Lactobacillus bulgaricus suspension. Availability: Items available for loan: UVAS Library [Call number: 2597-T] (1).
35.
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).
36.
Optimization Of Multiplex Pcr For Simultaneous Detection Of Bacterial And Viral Water Borne Pathogens
by Faiza Naz (2010-VA-288) | Prof. Dr. Masood Rabbani | Dr. Fareeha Akhtar | Dr. Sana Ullah Iqbal.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Waterborne illness is a serious issue throughout the world. Bacteria such as Salmonella spp., Shigella spp., E. coli spp. and from Viruses mostly Rotaviruses are involved in various waterborne outbreaks due to usage of contaminated water because of poor sanitation system mixing of waste material and fecal material with water, which can be transferred to human body by consuming such contaminated water Detection of these bacteria and virus from various foods by conventional method is not easy. Conventional methods are time consuming laborious and expensive. Now multiplex PCR is widely used for rapid detection of waterborne pathogens. The method is more sensitive and specific and can detect more than one pathogen in one single reaction mixture. This experimental design is developed to optimize the multiplex PCR reaction for detection of Salmonella spp., Shigella spp., Escherichia coli spp. and Rota virus.
ATCC culture of Salmonella spp., Shigella spp., Escherichia coli spp., revived using standard culturing technique and multiplex PCR is optimized to amplify four different microbial genes simultaneously. A total 100 samples obtained from 10 towns, of Lahore. The samples were processed for multiplex PCR for detection of E. coli spp., Salmonella spp., Shigella spp. and Rota virus directly from water samples.
With the amplification of 4 bacterial and viral genes simultaneously multiplex PCR was optimized. Water samples were obtained, to check the strength of planned experiment in the field. The samples were processed for and multiplex PCR for direct detection of Salmonella spp. and E. coli spp. directly from water samples.
Similarly multiplex PCR was optimized with 3μl DNA template of each microbe , 56oC annealing temperature , 20pmol of every primer and 25μl of multiplex master mix.
Multiplex PCR is more sensitive and specific. It is also time redeemable technique because conventional culturing method requires several days for the detection of waterborne pathogens but this technique wants expertise.
This study was helpful to establish an optimized Multiplex PCR for the rapid and simultaneous detection of waterborne pathogens. Availability: Items available for loan: UVAS Library [Call number: 2612-T] (1).
37.
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).
38.
Molecular And Serological Characterization Of Soilborne Francisella Tularensis
by Javed Muhammad (2010-VA-65) | Prof. Dr. Masood Rabbani | Prof. Dr. Khushi Muhammad | Dr. Muhammad Wasim.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Tularemia, caused by Francisella tularensis (F. tularensis ), is a zoonotic disease transmitted through contact with infected animals and contaminated environment. The disease has been reported from many countries of the world but no study has been done in Pakistan. In the current project, a total of 2280 soil samples representing 456 villages of eight districts of Punjab province were collected from way-points having human-animal interaction, processed for genomic extraction and tested through real time PCR for presence or absence of F. tularensis. Association of risk factors was determined from data such as gender and age of animals, plough method, irrigation system, fertilizer type used, availability of veterinary services, level of farmer education, physical and chemical composition of the soil. Moreover, sero-prevalence against F. tularensis in cattle, buffaloes, sheep and goats was determined using ELISA. Seventy four soil samples (3.24 percent) were found positive for F. tularensis. Phylogenetic analysis showed 100 percent similarity index with F. tularensis sub specie holarctica reported from other regions like USA, Sweden, Spain, Turkey and Germany. Presence of F. tularensis in soil showed negative association with increase in number of human density (0.7159; 0.3834-0.2054). Prevalence of anti- F. tularensis ELISA antibodies were significantly higher (p<0.05) in large ruminants (cattle and buffalo) as compared to small ruminants (goat and sheep). Age and gender-wise analyses showed non-significant differences (p>0.05) between small and large ruminants. Whereas, rain-irrigation system (2.96: 1.35- 6.48), lack of veterinary services (4.77:1.26-18.03) and use of organic fertilizer (5.3: 11.38- 20.39) have positive association with prevalence of anti- F. tularensis ELISA antibodies in the serum. Sero-prevalence of Ft in the animals has significant association with quantity of clay in soil (p<0.05). A conventional PCR based test has also been optimized for
Summary
103
detection of F. tularensis using tul4 gene specific primers. Specificity of primer showed F. tularensis detection in soil DNA in the presence of other cross-reactive organism. Sensitivity was determined in two fold dilutions with detection limit of up to 320 pg/μL. Utilizing pET28a vector, a construct was prepared containing transformed tul4 gene (450bp) showing 100 percent sequence homology to query gene sequence. For manufacturing diagnostic assays especially in developing countries where availability of BSL-3 facilities and positive control reagents is an issue, provision of tul4 gene based constructs in vector can act as positive control and biosafe to use.
It is recommended that similar studies may be done in other parts of Pakistan to have spatial distribution of F. tularensis all over Pakistan. In future studies, other sources of transmission like water, ticks and rodents may be considered with soil for complete analysis. Transportation of whole genome of F. tularensis has been prohibited by Russian government, ATCC and CDC, WHO is working on designing a complete protocol for transportation of this bacteria or genome to other countries. Under such situation, conventional PCR optimization can be done for diagnosis of F. tularensis and pET28+tul4 constructs, developed in this study, can be used as a PCR positive control reagents. Availability: Items available for loan: UVAS Library [Call number: 2640-T] (1).
39.
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).
40.
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).
41.
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
53
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).
42.
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.
Summary
71
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
Summary
72
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).
43.
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).
44.
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
Summary
63
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).
45.
Molecular Characterization Of Bovine Viral Diarrhea Virus (Bvdv) In Commercially Available Bovine Semen In Pakistan
by Muhammad Usman Sajjad khan (2010-VA-223) | Dr. Arfan Ahmad | Prof. Dr. Masood Rabbani | Dr. Mati-Ur-Rehman Khan.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: The production potential of livestock is showing some compromising trends. This may be due to multiple factors including management, nutrition, stresses, microbes etc. Among microbes, BVD virus has been identified during a study conducted on limited number of samples. The source of BVD virus may be carrier status of imported exotic animals or exotic bovine semen. In Pakistan, most of the farmers are availing services of artificial insemination for breeding purposes without knowing the status of BVD virus. Beside the imported semen, it is also being produced at various semen production units and among others, BVD viruses are the main obstacle in generating revenue by exporting quality semen. These issues are leading to a stage of losing trust of the local clients in purchasing substandard semen on one hand and on the other hand not meeting the export requirements of various client countries. This study aims for the detection and molecular characterization of BVD virus in commercially available bovine semen and the comparative status of BVD virus in the semen of local and exotic breeds of cattle. Furthermore, the comparative status of BVD virus in semen of cattle and buffalo will also be observed.
The hypothesis for the current study is that bovine semen used for artificial insemination is a source of BVD virus.
In this study, a total of 84 commercial semen straws were procured and processed. Among these, 45 samples (25 Govt. and 20 Private SPUs) originating from 5 breeds of cattle including Dhanni, Dajjal, Sahiwal, Red Sindhi and Cholistani were procured. Similarly, 39 samples (24 from Buffalos and 15 from Exotic Cattle) were processed. All the samples were tested for confirmation of BVD virus through PCR using BVD virus specific primers.
Summary
35
The samples if found positive for BVD virus were to be further processed for identification of genotype in semen samples. Furthermore, all the samples were also tested to calculate the bacterial load (TVC/mL) by Plate Method of Colony counting. All the samples were cultured on MH agar, and the colonies were counted after an incubation period of 24 hours at 37ºC
The data obtained was analyzed statistically by using Binomial and Chi Square Tests by using the software ‘IBM SPSS Statistics’ (v18)
Actual Data about the status and circulating genotype of the BVDV in the commercial bovine semen of local and imported bovines was obtained, along with the status of bacterial contamination of commercial semen available in Pakistan.
The study showed that 0% percent of the tested semen samples had detectable BVDV RNA..
The Bacterial contamination of the semen samples was also interesting in the sense that a wide range of Bacterial Load was found in the samples; ranging from 600 CFU/mL to 9250 CFU/mL. Overall the Government SPUs showed a higher contamination with an average Bacterial load of 1975.2 CFU/mL in cattle semen and 3424 CFU/mL in Buffalo semen. In comparison; private SPUs had an average load of 1842 CFU/mL for cattle semen and 2482 CFU/mL for Buffalo semen.
Imported semen had an average bacterial load of 1128 CFU/mL Availability: Items available for loan: UVAS Library [Call number: 2876-T] (1).
46.
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
Summary
77
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,
Summary
78
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).
47.
Prevalance And Distribution Of Soil-Borne Escherichia Coli O157:H7 In District Lahore Of Punjab Province, Pakistan
by Hiba Tabassum (2011-VA-421) | Prof. Dr. Masood Rabbani | Dr. Ali Ahmed Sheikh | Dr. Sehrish Faryal.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Salmonella spp. and Campylobacter spp. (Campylobacter coli and Campylobacter jejuni) are
recognized as the leading causes of bacterial gastroenteritis, followed by Shigella spp. and
Shiga toxin-encoding Escherichia coli (STEC).(Control and Prevention 2010).Shigatoxigenic
Escherichia coli (STEC) include Escherichia coli serotypes whose genomes
contain one or more Shiga toxin genes. STEC infections in humans can range from mild selflimiting
diarrhea to more severe disease, including hemorrhagic colitis and hemolytic uremic
syndrome (HUS).
Real-time PCR allows for quantification of the target. Real-time PCR perform better
than the standard culture-based assays to detect pathogenic organisms. In summary, work
load and work flow issues may dictate which system is best for different-sized laboratories
and test volumes.PCR assays to perceive the stx1 and stx2 genes are utilized by several
public health laboratories for identification and confirmation of STEC infection. Depending
on the primers used, these assays will distinguish between stx1 and stx2 (Zaki and El-Adrosy
2007). Assays even have been developed that verify the specific O group of an organism,
detect virulence factors such as intimin and enterohemolysin, and can differentiate among the
subtypes of Shiga toxins.
So there was need to analyze soil of to check the presence of Escherichia coli
O157:H7 for avoiding fatal diseases caused by it and there was also monitored soil chemistry
and its relation with bacterial growth specifically Escherichia coli O157:H7 because
Different soil composition and different environmental risk factors promoted presence of
Escherichia coli O157:H7 in soil.
Real Time-PCR technique was opted to detect Escherichia coli O157:H7 in the soil
from distant areas of district Lahore of Punjab, Pakistan. Soil samples from 10 per cent
Summary
53
villages were collected from this district and handled for genome extraction using
commercially available soil DNA extraction kit. After genome extraction, the samples were
keep running for Real Time-PCR at optimized conditions. The reaction was improved by
variations in standard concentrations of primers, probes, DNA, Taq-polymerase and sequence
of primers.
The dissemination of soil borne Escherichia coli O157:H7 was mapped in mentioned
district of Punjab, Pakistan using geographical co-ordinates recorded by GPS beneficiary.
Relationship of Escherichia coli O157:H7 with environmental factors,soil chemistry and
source of land irrigation (Canal, tubewell and rain or in combination), was resolved. Results of
present project were analyzed through SPSS.
The purpose of the research work was the understanding of occurrence and
distribution of Escherichia coli O157:H7 in district Lahore of Punjab province .It also threw
light on role of soil as a reservoir of Escherichia coli O157:H7, and association of this
infectious agent with various risk factors.
In this study depending upon the statistical analysis data, it was depicted that
Escherichia coli o157 H7 is present in soil although it can’t persist or survive. The prevalence
rate of Escherichia coli O157 h7 is 3.1% in 129 soil samples of Lahore. In case of villages it
is present in 2 villages of of 29.that shows it is 6.8% prevalent in villages.
The presence or absence of pathogen in relation to soil chemistry and seven potential
risk factors was determined through student t distribution (T-test). By observing p value of
variables of positive and negative sites it comes to know that there is no significant
association of these factors to the survival of Escherichia coli O157:H7 in soil.
Remaining all other analytes has not significant association with soil. But it can be
seen Escherichia coli O157 H7 has significant association with organic matters, phosphorus,
Summary
54
copper, cobalt, calcium, sodium, ferrous ion, potassium and sand form of soil ranging from
(0.86-1.97), (9.7-22.5), (24.18-41.12), (0.048-0.51), (0.39-0.96), (0.08-0.16), (41.84-59.14),
(51.02-69.56), (83-86) respectively. Remaining all other analytes has not significant
association with soil.
For further investigations it is necessary that find out those factors which cause
hindrance in survival of Escherichia coli o157 h7 in soil specifically and also search out
those factors which support Escherichia coli O157 h7 persistence in soil. Availability: Items available for loan: UVAS Library [Call number: 2935-T] (1).
