1.
Isolation, Characterization Of Chondroitin Sulphate And Its Efficacy In Osteoarthritis
by Humaira Majeed Khan | Prof. Dr. Muhammad Ashraf | Prof. Dr. Mansur-ud-Din Ahmad.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2012Dissertation note: Chondroitin sulphate (CS) and Glucosamine sulphate (GS) are two main components of articular cartilage. It is believed that these molecules slow down wear and tear of cartilage. Moreover, if administered exogenously as drugs, these may initiate synthesizing capacity of cartilage. Among these, GS promotes the formation and repair of cartilage, whereas CS promotes elasticity and prevent cartilage breakdown by inhibiting degradative enzymes. Concurrent use of both structural units of cartilage as drugs in osteoarthritis (OA) may lessen the progression of disease.
The present study was conducted to elucidate the chicken keel cartilage as an alternate and potential source for this endogenous component that may be used exogenously to repair or prevent damage to joints. Chicken keel cartilages were collected from healthy broilers. CS was extracted using MgCl2 solution (3M), dialyzed and digested with papain. The extracted material was purified by ethanol precipitation, centrifugation and then freeze dried. Proximate analysis of semi-purified polysaccharides revealed the presence of carbohydrates (65.49±0.10), crude protein (12.82±0.26), ash (11.12±.56), moisture (9.88±0.32) and fat (0.69±0.14). Fiber contents were found to be nil in the processed samples. Dimethylmethylene blue binding (DMMB) assay was performed for determination of percent contents of CS in extracted semi-purified samples and mean concentration was found to be 70.77±2.35. Semi-purified polysaccharides were further characterized by FTIR (Fourier Transform Infrared Spectrometer) technique and characteristic Peaks of CS molecules were recorded at 854, 854 and 853 cm-1 and then compared with spectrum of standard CS. Protein content being a major impurity in extracted samples was determined by Bradford method quantitatively (4.64±0.29). Two protein impurities having 77.8 and 50.5 kDa molecular weights were revealed by SDS-PAGE.
Efficacy of semi-purified CS from chicken keel cartilage, standard CS from shark source and GS, alone and in combination in experimental OA rat model was evaluated. To develop OA similar to spontaneous OA, 10mg papain/0.5mL (Sigma, Cat # P 3125) in buffered solution of 0.05 M sodium acetate pH 4.5 was injected intra-articularly in each right knee joint of fifty five albino rats (pre-anesthetized with anesthetic ether). Ten rats (n= 10) were injected with 0.5mL of normal saline (0.9%) in right knee joint that served as control group. Then from fifty five papain injected rats, twenty five were divided into five groups (n=5) for development and assessment of OA model (OA groups). Progression of disease was monitored by clinical scores, histopathological scores and concentration of CTX-II as biomarker in sera samples of experimental rats by ELISA using a commercial kit (serum preclinical CartiLaps ® ELISA kit) for control and OA groups (n=5) on day 0 (control group) and days 1st, 7th, 14th, 21st and 28th post papain injection (OA groups). Highest mean clinical score (10.38±1.1) was observed on 1st day and least on 28th day post papain injection i.e. 5.00±.34. Highest mean histopathological score and CTX-II concentration was recorded on 28th day i.e. 12.82±1.64 and 36.82±3.81. Values of clinical scores, histopathological scores and CTX-II concentration reached to maximum on 21st day and then sustained thereon. Second phase of experiment is comprised of evaluating and comparing the efficacy of extracted CS samples (chicken keel cartilages), standard CS (shark source) alone and in combination with GS. For this purpose, remaining five rats out of ten injected with normal saline intra-articularly served as control groups along with treated and non treated groups of experimental rats. Remaining thirty OA induced rats were divided into six groups (five rats /group). Group 1 (n=5) called non treated group received only placebo till 60th day and served as negative control group.
Treated Group 2 received GS alone, Group 3 CS (standard) and Group 4 were given extracted CS. Group 5 was treated with combination of GS plus CS (standard) and Group 6 with GS plus CS (sample). Doses of glycosaminoglycans (GAGs) were administered as 1.2g/kg/day CS and 1.5g/kg/day GS alone and in combinations. Drugs were offered early in the morning in bolus form with feed (10g) after overnight fasting while non-treated group received only placebo (without any drug).
Anti-arthritis activities of CS standard and extracted alone and in combination with GS were assessed clinically, analyzed statistically by using one way ANOVA. Level of significance (P<0.05) was recorded by using Duncan's Multiple Range (DMR) Post hoc Test. Mean scores of clinical, histopathology and CTX-II concentrations observed at 60th day in control rats (without OA) were 0.00, 0.00 and 2.55, respectively. OA induced untreated group showed mean score for clinical signs, histopathological scores and CTX-II concentrations 4.15, 12.24 and 36.70 and GS treated group 3.19, 3.96 and 6.12 at 60th day of treatment, respectively. For CS (standard), mean scores of clinical signs, histopathological lesions and CTX-II concentrations were recorded as 2.64, 2.44 and 4.48 and for CS (extracted) were 2.26, 2.28 and 4.40 in sera correspondingly at 60th day of treatment. The lowest mean values of clinical signs, histopathology and CTX-II concentrations in sera of treated group with standard CS plus GS were found to be 0.94, 0.94 and 2.62 followed by extracted CS plus GS treated groups 01.05, 1.27 and 2.74, respectively. Clinical, histopathological scores and CTX-II concentrations in group of rats treated with combinations were found to reverse the diseased condition after 60th days of treatment as the values were close to that of normal rats and far away from OA rats. It is concluded that extracted CS from poultry has comparable efficacy with CS standard from shark source alone and in combination with GS. Poultry by-product (keel cartilage) is found to be an alternate and cheap source for CS (chondroprotective agent) as compare to expensive, less available and religiously prohibited source for Islamic countries particularly.
Availability: Items available for loan: UVAS Library [Call number: 1544,T] (1).
2.
Detection Of Vivax Malaria And Under Local Climatic Conditions
by Sarwat naz | Prof.Dr. Azhar maqbool | Prof. Dr. Mansur ud din ahmad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2012Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2060,T] (1).
3.
Epidemiology Of Bovine Tuberculosis And Its Public Health Significance In Peshawar
by Irfan Khatak (2011-VA-562) | Dr. Muhammad Hassan Mushtaq | Prof. Dr. Umer Sadique | Prof. Dr. Mansur-ud-Din Ahmad | Prof. Dr. Muhammad Sarwar Khan.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: A cross-sectional study was carried out to determine the prevalence of bovine tuberculosis (bTB)
and associated risk factors in cattle and buffalo in Peshawar, Pakistan. Cattle and buffalo,
randomly selected from all four towns of District Peshawar were screened for bovine
tuberculosis using comparative cervical intradermal tuberculin test (CCIT). For obtaining data on
risk factors, socio-demographic condition, animal characteristics and management, interviewer
administered pretested questionnaire to animal owners. Multivariable logistic regression models
were used to measure association between risk factors and comparative cervical intradermal
tuberculin reactors. A total of 556 cattle and buffalo were screened for bovine tuberculosis. Out
of 556 animals screened, 5.75% (3.9-8.0%) were found positive. The prevalence was higher in
old animals (P= 0.001) as compared to younger animals. Prevalence also varied with source of
animal (either raised on farm or purchased), stay of animals at night (indoor or outdoor) and herd
size. Farmer’s knowledge about transmission of TB from animals to human as well as signs and
symptoms of TB was extremely low. Only 3.6% farmers correctly stated the combination of
three major symptoms of TB. Results of the study call for immediate intervention to control bTB
in animals as well as its transmission to human population. Furthermore, it is suggested to
emphasize on local epidemiology of bTB and husbandry practices of cattle and buffalo during
the control program.
To assess the presence of Mycobacterium bovis (M. bovis) in milk sold at retail shops and find
the knowledge, attitude and practice (KAP) about tuberculosis (TB) in the high risk M. bovis
contaminated milk consumers, milk samples were obtained from 92 milk shops and analysed for
presence of M. bovis. Data on socio-demographic characteristics and KAP about TB was
Summary
152
obtained from 800 M. bovis contaminated milk consumers. Mycobacterium bovis was detected in
8.7% (8/92) milk samples. Although 97.4% of the participants had heard of TB but only 39.6%
knew that cough lasts for more than 3 weeks was one symptom. Only 79.2% have awareness that
TB can be prevented and the most frequently stated (48.4%) method of TB prevention was good
nutrition. Participants believed that TB can be cured by prayers/ eating well (41.8%) and also by
herbal cures/ consulting Hakeem (35.7%). Mean knowledge score for the participants was 12.1±
2.47 out of maximum 22. Mean knowledge score varied significantly with ethnicity, level of
education and residential status (Urban vs rural). Overall knowledge about TB was low.
Therefore community’s health education focused on increasing knowledge of TB must be
initiated.
This part of study was conducted to determine the occurrence of active pulmonary tuberculosis
due to M. bovis in abattoir workers, butchers, livestock farmers and veterinarians and to
document the Knowledge and practices of these professional regarding bTB. The cross sectional
study included 141 abattoir workers, 317 butchers, 50 livestock farmers, 5 veterinary doctors and
3 veterinary assistants. Sputum samples were collected from those respondents who had chronic
cough that last for more than 2 weeks. Four out of 16 suspected abattoir workers and 1 out of 50
livestock farmers were found positive for M. bovis by Polymerase chain reaction analysis.
Duration of work as abattoir worker was found significantly associated (p<0.05) with occurrence
of zoonotic TB. The knowledge of abattoir workers, butchers, livestock farmers and veterinary
assistants regarding transmission of bTB from animal to human and symptoms of TB in human
was very low. Most of these professional did not use protective material/ techniques and are
considered at high risk of acquiring zoonotic tuberculosis. This study declares zoonotic
tuberculosis a critical public health issue especially for professionally exposed groups in
Summary
153
Peshawar, Pakistan and warrant immediate intervention for control of bovine and zoonotic
tuberculosis.
The last part of study aims to determine the proportion of zoonotic TB cases out of overall
human TB patients and school children, drug resistance of M. bovis isolates and knowledge,
attitude and practices about TB. Total 300 human TB patients and 100 school children were
included in the study. Sputum samples were processed by PCR for presence of Mycobacterium
tuberculosis and M. bovis. Sputum samples from TB patients were cultured and M. bovis isolates
were subjected to drug susceptibility testing. Data on knowledge, attitude and practices were
obtained from TB patients by administering pre-tested questionnaire. Among TB patietns 4%
(12/300) were infected with M. bovis. None of the school children was positive for M. bovis.
Residence, occupation, presence of animals at home and sleeping in shed at night was found
significantly associated with occurrence of zoonotic TB. Except one all M. bovis isolates were
resistant to Pyrazinamide. Among other drugs resistance to streptomycin and isoniazid was high.
Low level of knowledge and practices were observed. The study concluded that considering
zoonotic aspect of TB during diagnosis and treatment of TB is necessary and recommends
national survey for true estimation of burden of zoonotic TB in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 2540-T] (1).
4.
Epidemiological Investigation of rRisk Factors of Mastitis in Dairy Buffalo and Antibiotic Resistance of Staphylococcus Aureus
by Abid hussain2010-va-261 | Prof. Dr. Mansur ud din ahmad.
Material type: Publisher: 2015Dissertation note: Phd. Availability: No items available
5.
Spatial Ecology And Distribution Of Soil Borne Burkholderia Mallei In Punjab, Pakistan
by Muhammad Asad Ali (2002-VA-73) | Prof. Dr. Khushi Muhammad | Prof. Dr. Aftab Ahmad Anjum | Prof. Dr. Mansur-Ud-Din Ahmad.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Burkholderia mallei is a causative agent of glanders, the disease of equines. The disease is characterized by pulmonary, nasal and cutaneous forms. B. mallei is excreted through nasal discharge, lacerated skin/wounds and expiration. Diseased animals shed bacteria through the discharges contaminating soil, water, fodder and other susceptible animals in its vicinity. The present study was designed to map and investigate the association of different physical factors and soil chemistry analytes with persistence of B. mallei genome in soil of 10% percent villages (n=456) from eight selected districts of Punjab province, Pakistan.
Eleven (0.48%) out of 2, 280 soil samples were positive for B. mallei genome in varied locations of Punjab. Higher prevalence (2.37%) for genome was detected in Sheikhupura district followed by Chakwal district (2.10%). None of the samples from Gujranwala, Sahiwal, DG Khan, Attock, Faisalabad and Sargodha districts were found positive for B. mallei genome. The genome of B. mallei was distributed in 25% study districts of Punjab, Pakistan. In Chakwal district, the genome of B. mallei was strongly associated with moisture (p=0.008) in all positive samples ranging from 0.80 to 39.20%, Phosphorous (p=0.050) ranging from 1.74 to 21.75 mg/Kg. While, this association in Sheikhupura district soil samples was with Sodium (p=0.018) and moisture (0.026) ranging from 1.90 to 133.59 mg/Kg and 0.80 to 39.20%, respectively. The odds of detecting DNA of B. mallei were recorded higher (1.4, 6.8, 5.0, 2.8 and 10.6 ) when soil sample sites were < 500 meters away from vehicular traffic roads, < one kilometer from animal markets, < 100 meters from canal, animal density < 1,000 animals and human population < 300 houses/village. While the odds of detecting DNA of B. mallei were 0.1, 0.3, 0.4, 0.2 and 0.5 when soil sample sites were > 500 meters from vehicular traffic roads, > one kilometer from animal markets, > 100 meters from canal, animal density > 1000 animals and human population > 300 houses/village, respectively. Soil-borne B. mallei DNA is more likely to be detected in areas closer to roads with vehicular traffic along the interstate routes in Punjab and soil containing low level of moisture.
It was concluded that soil of two districts out of eight selected was positive for B. mallei genome in Punjab province. Odds of less distance from main road to animal farm and high animal density at farm were positively associated with B. mallei DNA persistence in soil. Moisture, sodium and phosphorus were positively associated with persistence of B. mallei DNA in soil.
Availability: Items available for loan: UVAS Library [Call number: 2900-T] (1).
