1.
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).
2.
Physico-Chemical Factors Affecting The Growth Of Bovine Rotavirus
by Wardah sharmeen syed | Prof. Dr. Tahir yayub | Dr Muhammad Zubair shabbir | Dr.Muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1924,T] (1).
3.
Prevalence And Risk Analysis Of Coxiella Burnetii In Soil Of Faisalabad And Gujranwala Districts
by Zia Ul Hasnain (2007-VA-290) | Dr. Muhammad Zubair Shabbir | Dr. Arfan Ahmed | Dr. Muhammad Yasir Zahoor.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Coxiella burnetii is a causative agent of Q-fever, a widespread zoonosis. The effective adaptation of C. burnetii to intracellular existence is in contrast with its ability to survive in the environment outside the host cells and its resistance to chemical and physical agents. Besides nutrients and minerals, soil is aggregate of number of pathogens. Many of those organisms are of zoontoic importance and have significant threat to public health. One 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. Soil borne C. burnetii has not shown any association with different risk factors. The factors include presence or absence of pathogens with or without animal interaction, distance from animal market, main road, canal, animal and human density in a village under study. PCR technique was used to identify C. burnetii in the soils of Faisalabad and Gujranwala district. Soil samples (n= 730) were collected from each village of the both districts and processed for genome extraction using commercial soil DNA extraction kit. The extracted DNA from the soil samples was run further for PCR analysis of transposase IS1111a followed by standard gel electrophoresis technique. Only 6 (0.82%) samples were positive out of 730. Furthermore pathogens prevalence was geographically mapped in relation to roads, canals,
-----------------------------------------------------------------------------------------------------------------------Summary
45
rivers, drains, animal and human population to determine the risk areas according to intensity of identified pathogens for both districts. Odd Ratio was calculated to access the association in terms of absence or presence of pathogens with particular risk factors, which did not show any kind of association between pathogen and risk factors. The phylogenetic analysis of C. burnetii shows different convergence percentage with the isolates of worldwide i.e., Namibia (99.53%), Brazil (100%), Taiwan (99.53%), India (99.07%).
Study was contributed to understand about the previously unrevealed prevalence of C. burnetii in soil of district Gujranwala and Faisalabad together with risk factor analysis implicating possible health significance as well as survival in the soil. Availability: Items available for loan: UVAS Library [Call number: 2312-T] (1).
4.
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).
5.
Isolation And Antibiotic Susceptibility Pattern Of Extended Spectrum Β-Lactamases Producing Klebsiella Pneumoniae From Human Clinical Specimen
by Muhammad Tahir Ishaq (2013-va-443) | Dr. Jawad Nazir | Dr. Muhammad Zubair Shabbir | Dr. Aqeel Javeed.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Klebsiella species particularly K. pneumoniae are important opportunistic nosocomial pathogens causing a variety of infections including urinary tract infections (UTIs), pneumonia, septicemia and wound infections. Epidemic and endemic nosocomial infections caused by K. pneumonia species are leading causes of morbidity and mortality throughout the world. Irrational use of antibiotics is creating antibiotic resistance problems in Pakistan. Excessive use of β-lactam antibiotics is responsible for production of ESBL enzymes by Gram negative bacteria especially K. pneumoniae.
A total of 150 samples including urine, pus and sputum were processed for the isolation and evaluation of the ESBL enzyme producing K. pneumoniae as well as their antimicrobial sensitivity pattern. Samples were cultured on cystine lactose electrolyte deficient (CLED), blood, chocolate and McConky agar. Isolates were identified by culture characters, staining reaction followed by biochemical testing through API-20E index system. ESBL production potential was assessed by double disc diffusion method. Out of total 50 urine samples, K. pneumoniae was isolated from 20 samples. Among these 20 isolates, 6 were confirmed as ESBL producers. However, within 50 sputum and pus samples, 13 and 15 isolations were possible out of which 5 and 6 were positive for ESBL production, respectively. Frequency distribution for isolation of K. pneumoniae from urine, pus and sputum samples did not significantly vary from each other. Similarly, ESBL producing potential of all K. pneumoniae isolates also did not significantly vary as p values are higher than 0.05.
A total of 17 ESBL producing K.pneumoniae isolates were tested for their antibiotic susceptibility pattern against 12 commonly used antibiotics i.e. Amoxacillin, Ampicillin, Aztreonam, Ceftazidime, Cefatoxime, Ceftriaxone, Cefixime, Imipenem, Meropenem, Ciprofloxacin, Gentamycin and Amikacin. All of the tested K. pneumoniae isolates showed 100 % sensitivity against amikacin, imipenem and meropenem while found to be completely resistant against rest of the antibiotics.
Molecular confirmation of ESBL production was done through PCR. DNA samples were extracted from ESBL producing isolates. Amplification of the plasmid DNA region (TEM gene) from the DNA samples was done through PCR. The resulting PCR product was run on 1% agarose gel through horizontal gel electrophoresis. Three samples from urine (S1, S4 and S5) and two samples from sputum (S7 and S8) produced required bands while none of the other samples produced any band.
High proportion of ESBL producing K. pneumoniae isolates in present study is an alarming scenario. These organisms are resistant to conventional antibiotics and might spread in the environments thus pose serious threat to the health of human and animal population. Only clinical specimen submitted to the diagnostic lab were tested in present study. Further extensive studies are needed to bridge the gaps in knowledge of antibiotic resistance pattern of K. pneumoniae. Availability: Items available for loan: UVAS Library [Call number: 2366-T] (1).
6.
Detection Of Amantadine Resistant Variants Among Avian Influenza Viruses Subtype H9n2 Isolated In Pakistan
by Sabir Subhan (2009-VA-32) | Prof. Dr. Tahir Yaqub | Dr. Muhammad Zubair Shabbir | Dr. Yasin Tipu.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Avian influenza A virus subtype H9N2 is prevalent in poultry industry of Pakistan. Amantadine is an antiviral drug which is being used as prophylactic measure to control this disease despite the occurrence of resistance against this drug. There is need to monitor the resistance of amantadine in Flu viruses. In this study, we collected 100 samples of broilers birds showing mild to severe respiratory signs. Samples were collected from different locations of Punjab, Pakistan. After initial identification via Hemagglutination test, the molecular identification and confirmation of subtype H9N2 was done by multiplex PCR. To rule out the co-infection of NDV, PCR of NDV was also done. The samples which were pure H9N2 were further processed for the screening of amantadine susceptibility. To do this, titration of viruses was done on MDCK cells in the presence and absence of amantadine at the concentration of 2 ug /ml. The results of TCID5O were compared in the presence and absence of amantadine for each isolate and isolates showing difference of 2 log 10 TCID50/0.1 ml were declared resistant to amantadine as described by Masuda et al. 2000. The results of this study revealed that the viruses circulating in the poultry industry if Pakistan are resistant to this drug as we found that out 10 isolates 4 were resistant to this drug. So, there is need to monitor the usage of this drug in poultry as human cases of H9N2 viruses have been reported and virus was of avian origin. Monitoring is necessary because amantadine is recommended in flu pandemics and this virus possesses the pandemic potential and can cross the species barrier. Availability: Items available for loan: UVAS Library [Call number: 2457-T] (1).
7.
Seroprevalence And Risk Factor Analysis Of Bluetongue Virus In Lahore And Faisalabad Districts Of Punjab Province, Pakistan
by Syeda Marriam Maqbool (2014-VA-522) | Dr. Muhammad Zubair Shabbir | Dr. Ali Ahmad Sheikh | Dr. Maryam Javed.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Domestic animals play an important role in the rural and agricultural economies of developing countries. Therefore, animal diseases pose a threat to country’s economy, animal welfare, the environmental and public health. One of the important animal diseases is Bluetongue, listed as notifiable disease by OIE. Causative agent is Bluetongue virus (BTV) an arbovirus that belongs to genus Orbivirus with in family Reoviridae. The main route of transmission is through the bite of Culicoides biting midges. Disease is enzootic and widely distributed in areas where susceptible animals and vector species are prevalent. It has been reported worldwide including the neighboring countries of Pakistan. BTV is also considered an endemic in Pakistan but little information is available on its epidemiology in this area. Serological tests can detect antibodies produced against infection and helpful to analyze the prevalence of a pathogen in circumstances when there lacks vaccination practice to ruminants in a given geographical area.
Competitive ELISA was used to identify antibodies to BTV in the sera samples of animals in Faisalabad and Lahore districts. Blood samples were collected from randomly selected villages of both districts and processed for serum separation by using gel/clot activator tubes. Separated serum was analyzed by competitive ELISA. Further, statistical analysis was done by OpenEpi to check the association between BTV seroprevalence and potential risk factors. Later the BTV prevalence has been mapped in relation to different villages of both districts.
Results of present study revealed that Bluetongue virus is prevalent in Faisalabad and Lahore districts with high seropositivity observed for Faisalabad district. Antibodies to BTV were detected in all studied animals irrespective of their age, sex, parity and breed. Risk factor analysis is implicating the association of BTV seroprevalence with breed, sex and age for sheep,
SUMMARY
44
cattle and buffalo respectively. Further studies should be conducted to expand the geographical area for the assessment of Bluetongue prevalence and to explore the genetic diversity of Bluetongue virus. Availability: Items available for loan: UVAS Library [Call number: 2497-T] (1).
8.
Isolation, Identification And Antibiotic Resistance Profile Of Bacterial Isolates Of Public Health Significance In Raw Milk Of Cows And Buffaloes
by Hafiza Khadija Naseem (2010-VA-315) | Dr. Arfan Ahmad | Dr. Muhammad Zubair Shabbir | Dr.Muhammad Hassan Mushtaq .
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Milk is a highly nutritious food that can be obtained from a variety of animal sources such as cows, goats, sheep and buffalo for human consumption. On account of zoonotic importance of some opportunistic pathogens of public health significance in milk, the studytherefore was designed for isolation identification and antibiotic resistance assessment of Staphylococcus aureus, Escherchia coli and Salmonellaspp in raw milk of cows and buffaloes.
A total of 60 raw milk samples were collected from buffaloes (n=30) and cows (n=30) located in an area around district Lahore. To evaluate the source of milk borne pathogens in milk, half samples (n=15) of each animal species were taken directly from udder while remaining half (n=15) from milking utensils at the same farm. Samples were cultured for isolation and confirmed by biochemical tests and their antibiotic resistance pattern was checked by Kirby baur disk diffusion test.
Samples that were taken from udder of buffalos,E.coli, Salmonellaspp and Staph aureus was isolated from 60%, 26% and 46% of processed samples whereas from utensils E.coli, Salmonella spp. and Staph aureuswas isolated from 66.66%, 66.66% and 73.33%of samples respectively.
While Samples that were taken from udder of cow’sE.coli, Salmonella and Staph aureus was isolated from13%, 0% and 40% of processed samples where as samples that were taken from utensils at the same farm E.coli, Salmonella and Staph aureuswas isolated 26%, 26.66% and 46% respectively.
Antibiotic resistance pattern of isolates E.coli, Salmonellaspp and Staph aureus showed 100% resistance to Lincomycin and Tylosine. Ciprofloxacin showed 75% sensitivity followed by Ciprofloxacin showed 75% sensitivity followed by 65%Oxytetracyclin 60% doxycycline and 58% Amoxicillin.
CONCLUSION
Our study reveals that raw milk supplied and consumed in Lahore city of Pakistan is contaminated with public health significancebacteria Salmonella spp, E.coli and Staph aureusdue to unhygienic conditions and milking practices.Data ofAntibiotic resistant profiling of these isolates showed 100% resistant to Lincomycinand Tylosine. While sensitive against Ciprofloxacin> Doxycycline >Oxytetracycline.Efforts should be made to use antibiotics wisely and hygienic practices should be followed during collection to supply chain of milk to avoid spread of antibiotic resistant bacterial strains from animal source to human beings.
Availability: Items available for loan: UVAS Library [Call number: 2521-T] (1).
9.
Study On The Status And Risk Factors Of Brucellosis In Bovines Of District Poonch, Azad Jammu And Kashmir
by Muhammad Kashif Idrees (2008-VA-68) | Dr. Arfan Ahmad | Dr. Muhammad Zubair Shabbir | Dr. Muhammad Avais.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Brucellosis is one of the main diseases which played a blemished role in destroying the economy of livestock farmers in Azad Jammu and Kashmir in the form of low productivity and reproductive disorders. In district Poonch, abortions rates in bovines have been increased tremendously during the last decade. To find out the Status of brucellosis in cattle and buffalo, 300 animals (n=150 cattle; n=150 buffalo) were randomly selected and screened for brucellosis in district Poonch AJK. Various risk factors like species, age, sex, pregnancy, lactation, abortion, breeds, repeat breeding, retained placenta, housing, feeding, management were also evaluated for their impact on occurrence of brucellosis in this area. Data regarding risk factors of each animal was recorded in a Performa (Attached as annexure A). Serum samples were collected from these animals and analyzed through RBPT. The serum samples positive for Brucella abortus through RBPT were further subjected to indirect ELISA for further confirmation. Serum samples analysis was done at University Diagnostic Lab, University of Veterinary and Animal Science, Lahore. The results showed the overall seropositivity of 3.7% and 2.7% in cattle and buffalo through RBPT and i-ELISA, respectively. Moreover the result revealed that in cattle the positivity was more (4.6%) and in buffalo was comparatively less (2.7%) through RBPT and through i-ELISA same more positivity (3.4%) in cattle than buffaloes (2.0%). The results revealed that positivity of brucellosis increases with the age. The positivity in non-pregnant was more than that of pregnant while the positivity in non-lactating animals more than the lactating animals. The animals with the history of abortion and retained placenta were more serological positive than the animals without history such history. In breed wise comparison crossbreed cattle and Nili-Ravi buffaloes evidenced more serological positive
43
Summary
percentage. Sex wise the female animals have more positivity compared to male animals. As far as village wise positivity Hajera, Davarandi, Mandol, Madarpur, Nakkar were evidenced for presence of brucellosis. Regarding impact of risk factors in the occurrence of brucellosis, statistically there was non-significant (≥0.5) difference observed in this study. The findings of this study evidenced that brucellosis is present and endemic in cattle and buffaloes in the district Poonch. However it was suggested that more surveys are required across the country in order to formulate a policy for prevention and control of brucellosis in livestock. Availability: Items available for loan: UVAS Library [Call number: 2625-T] (1).
10.
Histopathological And Biochemical Evaluation Of Chemical Castration In Rabbits
by Hadia Uzair (2010-VA-205) | Prof Dr. Zafar Iqbal Chaudhry | Dr. Ghulam Mustafa | Dr. Muhammad Zubair Shabbir.
Material type: Publisher: 2017Dissertation note: Overpopulation of companion animals accounts for millions of deaths, billions of spending and hundreds of serious bites to humans each year. In order to cope this over growing population of stray animals, several sterilization programs have been devised.
Rabbits will be subjected to intra-testicular 10% and 20% calcium chloride solution. Clinical observation and testicular volume measurement will be done on weekly basis throughout the study. Blood and testicles (by orchiectomy) will be collected after every 10 days, hematology and histopathology be done thereafter. Serum testosterone would be quantified by using radioimmunoassay to assess testicular function according to standard protocol.
In our study, the efficacy of injecting intra-testicular calcium chloride solution in alcohol, was compared for chemical-sterilization in 24 adult rabbits. 10% and 20 % solution of calcium chloride were administered, intra-testicularly in testicle bilaterally which were removed, with the open technique surgically after 30 days, these harvested testicles are than evaluated histopathologically. Serum testosterone was quantified by using radioimmunoassay to assess testicular function according to standard protocol .Blood picture of the rabbits was also observed for any clinical and subclinical complication.
Swelling of testicles was marked in both groups following injection of 10% and 20% calcium chloride and within 48 hours swelling reached to its maximum level. Though volume of the testicles reduced significantly treated group after three weeks of treatment. Treated testicles with calcium chloride underwent atrophy at the 30th day in studied experimental group, with no noticeable modification in control group. Testosterone level dropped significantly even after 15 days of post injection and on 30th day testosterone activity seems to be diminished.
Summary
62
The method is considered as applicable with no major adverse effects in general health of the animal. Results were considered satisfactory and this method can be applied as mass scale particularly, where the feasibility of surgical castration doesn’t exist. Extensive necrosis, sloughing off epithelium, infarction following fibrosis of tissue, shrinkage and germ cell apoptosis are presumed to be due to calcium chloride. In our study; severe diffuse necrosis of tubular structure along with progressive degrees of inflammatory response were observed as a main finding Availability: Items available for loan: UVAS Library [Call number: 2851-T] (1).
11.
A Thesis Submitted In The Partial Fulfillment Of The Requirements For The Degree
by Ayesha Saddiqa (2011-VA-367) | Dr. Asif Nadeem | Dr. Maryam Javed | Dr. Muhammad Zubair Shabbir.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2017Dissertation note: AIDS epidemic is increasing rapidly in the Eastern Mediterranean Region. Quoting fresh authorized figures collected by the Punjab health department were 97,000 to 125,000 HIV positive people in Pakistan. Number of patients with HIV/AIDS rapidly increased in Punjab. 310 HIV/AIDS cases (35 women and 13 children) have been stated in Punjab in 2014. CCR5 gene is associated HIV infection. Mutation in this gene delayed the progression towards AIDS. In this study blood samples were collected from the laboratory of Punjab Aids Control Program (PACP), primary and secondary health care department, Government of Punjab. Genomic DNA was extracted by using the using FavorPrepTM Blood/Cultured Cell Genomic DNA Extraction Mini Kit. Specific set of primers were designed for the amplification of the targeted gene. The amplified PCR products were precipitated and sequenced for the identification of polymorphisms. Bidirectional sequencing was done for result confirmation. Alignments of sequences were done with the help of NCBI BLAST. Chromas software, Clustal W, UCSC, Bio Edit and SNPedia and Mega 6.0 was used to compile this study. CCR5 32 base pairs allele deletion was found absent in all HIV positive and negative individuals. So, susceptibility of human immuno-deficiency virus type one infection is high in Pakistani population. Genomic comparison was done with non-human primates. Alignment result showed human CCR5 gene homology, 95%, 99%, 94% and 94% with Maccaca mulata (Rhesus Monkey), Pan troglodytes (chimpanzee), Cercocebus atys (sooty mangabey) and Rhinopithecus bieti (black snub-nosed monkey) respectively. So, this homology analysis showed that these non-human primates can be used for development of therapeutic strategies related to human immune deficiency virus. Availability: Items available for loan: UVAS Library [Call number: 2872-T] (1).
