1.
Chemical Equivalence Of Different Brands Of Oxytertacycline Hydrochloride And Its Minimum
by Sadaf Hina | Prof. Dr. Muhammad Ashraf | Dr. Aftab | Dr. Muhammad Adil Rasheed.
Material type: ; Format:
print
; Nature of contents: ; Literary form: Publisher: 2012Dissertation note: This project was designed to study the chemical equivalence of various brands of Oxytetracycline hydrochloride (long acting, short acting & PVP) approved by the ministry of health and available in the local market for veterinary use. Oxytetracycline was measured by HPLC method developed and standardized in the laboratory. Limit of detection (LOD) and limit of quantification (LOQ) of the Oxytetracycline by HPLC assay method were determined. From stock solution of working standard (Oxytetracycline hydrochloride) different concentrations 0.05, 0.1, 0.5, 1.0, 10, 25, 50 and 100µg per ml were prepared for the determination of LOD. The LOD calculated was 0.100(µg/ml) and LOQ was 0.5 (µg/ml). Correlation coefficient was 0.99994050. Concentration of the active ingredient (Oxytetracycline hydrochloride) in all preparations was same as mentioned on the label except Oxytetracycline (74%), Terrasym PVP-100 (81%), and Onyx-LA (72%).
MIC of Oxytetracycline hydrochloride against following bacterial isolates determined by micro-broth dilution test was Bacillus subtilis (50µg), Staphylococcus aureus (100µg), Eschericiha coli (50µg), Salmonella enterica (1000µg) and Pasturella multocida (50µg).It showed that all these bacterial cultures have developed resistance against Oxytetracycline hydrochloride.
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2.
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.
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3.
Toxicity Problems Associated With Declofenac In Avaian Species And Its Substitute
by Muhammad Ramzan | Dr. Haji Ahmad Hashmi | Dr. Zafar Iqbal Chaudhry | Prof. Dr. Talat.
Material type: ; Format:
print
; Literary form:
drama
Publisher: 2010Dissertation note: A catastrophic decline in vulture populations was first observed in 1996-97 in Keoladeo National Park, Bharatpur, Rajasthan, India. Later, similar situations were reported in many south Asian countries including Pakistan, Nepal and Bangladesh. Now, the International Union for the Conservation of Nature (IUCN) has listed three vulture species i.e. Gyps bengalensis, Gyps indicus and Gyps tenuirostris as 'critically endangered'.
Vultures are natural scavengers and play a key role in keeping the environment clean by consuming carcasses of dead livestock and wildlife. The unconsumed animal carcasses pose a serious threat to both human and animal health because decaying animal carcasses may contaminate groundwater and become a potential source of diseases such as tuberculosis and anthrax for humans and other animals. Also, due to the declines in vulture populations, a rise in the numbers of other scavengers like dogs has been observed. Therefore, the risk of dog bite and transmission of dangerous diseases, including rabies, has also increased. Furthermore, vultures play a vital role in the disposal of human corpses of the followers of the Parsi religion because they place their dead bodies before vultures for disposal rather burial beneath the earth.
In the South Asian region, different communities have different attitudes toward the use of meat and its products. For example, Muslims do not use meat of dead animals, whereas Hindus abstain from cow's meat altogether because of religious bindings. For many centuries, the disposal of such meat has been done by vultures.
The vulture population decline issue was investigated by researchers with the support of various international organizations dedicated to bird conservation in collaboration with regional ornithological societies. Scientists looked into the problem from different angles and considered a variety of reasons of this decline. These included food shortages, losses of habitat, persecution, human disturbances, infectious agents, environmental contaminants, intentional poisoning, and accidental poisoning through food or water. Initially, the outbreak of some infectious disease and/or poisoning appeared to be the most plausible basis of this crisis. Recent studies have ruled out presence of a widespread infection and have focused on some form of poisoning. Now, it is believed that diclofenac, a veterinary drug, was responsible for the huge fatalities in vultures, in part because the onset of the crisis was coincident with the introduction of this drug in veterinary practice.
Diclofenac belongs to a class of drugs called, non-steroidal anti-inflammatory drugs (NSAIDs). Diclofenac served as an effective analgesic (pain killer), antipyretic (reduces fever), and anti-inflammatory (reduces swelling) drug. Initially, this drug was used in human beings for various indications such as arthritis. The use of diclofenac was started in domestic animals in the region a decade ago. It was reported that the presence of diclofenac in the bodies of dead animals that had been treated with this drug shortly before death was harming the vultures feeding on contaminated carcasses (Oaks et al., 2004; Shultz et al., 2004; Swan et al., 2005).
The Department of Pharmacology and Toxicology, at the University of Veterinary and Animal Sciences, Lahore, has been given the task to investigate this problem. A series of experiments was conducted to study the likely causes of this problem and, if possible, find its solution by developing safer and efficacious alternatives to diclofenac for the treatment of animals. For this purpose, a chicken experimental model was developed to study the toxicity of diclofenac and other NSAIDs. These studies have shown that diclofenac produced similar toxic effects and mortalities in broiler chickens as had been reported from studies of diclofenac-poisoned vultures. Later on, therapeutic efficacy studies of safer alternative NSAIDs of diclofenac were conducted in horses, buffaloes.
On the basis of these current studies, it was concluded that diclofenac was toxic to chickens and no significant difference was present in the death rates in bird groups treated with toxic doses of diclofenac via oral and intramuscular routes. Sodium and potassium salts of diclofenac caused comparable casualties in broiler chickens (unpublished study). A number of other NSAIDs were screened for their toxicity profile using the chicken model. This study showed that phenylbutazone, dipyrone, meloxicam, piroxicam and ketoprofen were comparatively safer than diclofenac. However, phenylbutazone and dipyrone may not be appropriate alternatives for diclofenac in food-producing animals because they are known to cause a condition called agranulocytosis in human and animals. Agranulocytosis involves marked reductions in numbers of white blood cells that are responsible for maintaining the body's immunity against various diseases.
To evaluate the efficacy of safer drugs, fever was induced in buffalo calves with Escherichia coli endotoxin (lipopolysaccharide), and the animals were then treated them with ketoprofen, meloxicam orpiroxicam. Although, all three drugs were effective in lowering body temperature, ketoprofen was the most efficient. In another experiment, these drugs were used in the treatment of lameness in horses, and it was concluded that meloxicam was more effective followed by piroxicam and ketoprofen for the treatment of this particular problem.
Based on these observations, it is concluded that ketoprofen, meloxicam and piroxicam may prove quite safe drugs for the scavenging birds and may be used as safe alternatives to diclofenac in veterinary practice. It has been observed during this study that ketoprofen, piroxicam, and meloxicam are less toxic for broiler chickens and may prove better alternates to be used in place of diclofenac in animals. The use of these NSAIDs may be less toxic for scavenger birds. The commercial preparations of ketoprofen and meloxicam for veterinary use are available at least in two south-east Asian countries i.e. Pakistan and India. The prices of these NSAIDs are comparable with those of the veterinary preparations of diclofenac which were previously available in the region. On the basis of results of this study it is concluded that ketoprofen, piroxicam, and meloxicam are effective NSAIDs in domestic animals, hence may replace diclofenac in veterinary practice.
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