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1. Etiological Study Of Pancytopenic Children

by Dr. Syed Maaz Nadeem | Dr. Muti-ur-Rehman Khan | Dr. Aftab Ahmed Anjum | Dr. Asim Aslam.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Pancytopenia is a hematological condition in which there is a decrease in all three cell lines of peripheral blood i.e. erythrocytes, leucocytes and platelets leading to anemia, leucopenia and thrombocytopenia. Complications of anemia, repeated infections and bleeding tendencies are sometimes horrifying and may result in death of individual. The present study was designed to analyze the underlying pathology, different clinico-haematological features and importance of bone marrow study in one hundred children presenting with pancytopenia. Present study was carried out in pediatric laboratory of Mayo Hospital, Lahore, Pakistan.. Detailed history was taken in all cases. Complete blood counts were done on an automated blood analyzer (Sysmex Kx-21). For counter check of total leucocyte count, differential leucocyte count and platelets, smears were also prepared and stained by using Giemsa stain. Red cell morphology was done on blood smear for theconfirmation of red cell indices. A total volume of 3 ml venous blood was drawn into a syringe. Out of which 1.0 ml was delivered into EDTA containing vacutainer and remaining 2 ml blood was transferred to a plain glass tube. After clotting and centrifugation serum was obtained for screening of hepatitis B surface antigen and antibodies against hepatitis C virus. Bone marrow aspiration was also performed where indicated. Megaloblastic anaemia (42%) Aplastic anaemia (26%) and ALL (8%) were found to be the common causes of pancytopenia in our scenario. Less common causes of pancytopenia were infections (8%), mixed deficiency (4%), MDS (4%) and lymphoma (4%). In all above mentioned cases clinical manifestations and peripheral blood counts played an important role in their evaluation. Two cases of haemophagocytic syndrome (2%), a rare cause of pancytopenia were also diagnosed in this study. This study also explained the importance of physical examination, peripheral blood findings and bone marrow examinations in the management of pancytopenic patients. Peripheral blood film and bone marrow aspiration should be performed simultaneously in pancytopenia patients when the diagnosis is not confirmed. Bone marrow examination in most cases gives the specific diagnosis. However, in few cases, additional tests may be required. Serum vitamin B12 and folic acid levels may be needed for confirmation of megaloblastic anemia. Serum iron, TIBC and iron staining on bone marrow smears may be required in iron deficiency anemia. In cases of leukemia flow cytometry study may be more helpful in reaching a final diagnosis. Bone marrow biopsy is mandatory in aplastic anemia. Availability: Items available for loan: UVAS Library [Call number: 1539,T] (1).

2. Patho-Morphological Studies Of Reproductive Tract Disorders In Cattle And Buffalo

by Abdul Jabar | Dr. Ishtiaq Ahmed | Dr. Aftab Ahmed Anjum | Dr. Asim Aslam.

Material type: book Book; Format: print ; Literary form: drama Publisher: 2012Dissertation note: Reproductive tract disorders like endometritis, pyometra, follicular cysts, leuteal cysts etc have been reported as major causes of low conception rate and poor productivity of livestock. Reproductive tract lesions in cattle and buffaloes play an important role in animal breeding either by causing infertility or sterility of animals resulting in high economic losses. Besides of other reasons, presence of different types of bacteria inside the uterus of breeding animals causes deleterious effects on the reproductive tracts causing severe disorders and ultimately leads to failure of pregnancy. All these microbes results in the infection of uterus that ultimately affects animal's conception rate. A total 100 reproductive tracts of slaughtered cattle (n=40) and buffaloes (n=60) were randomly collected from Lahore abattoir, Sherakot to study the impact of bacteria on the reproductive tracts, efficacy of variety of antibiotics on different bacterial isolates, gross and histopathological lesions. The swab samples for bacteriology were processed for purification and identification of bacterial isolates by doing number of conventional tests for final characterization and in vitro antibiotic sensitivity was performed. In this study seven different bacterial isolates were identified from all the samples. These include: Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Corynebacterium species, Aeromonas species, Micrococcus species and Lactobacillus species. By the analysis of in vitro antibiotic sensitivity, all eight different species of bacteria isolated were 100% sensitive to Ciprofloxacin, Norfloxacin and Chlortetracycline. Contrary to these, all bacteria were 100 % resistant to Bacitracin followed by Sulphaphenazole, Ticarcillin/Clavulanic acid and Trimethoprim. Out of the total 100 reproductive tracts of cattle and buffaloes collected randomly, 54 showed lesions with different severity. Most of the lesions were uterine hemorrhages followed by pyometra, para-ovarian cysts, pyosalpinx, hydrosalpinx, leuteal cysts, follicular cysts and uterine tumors. These lesions were observed slightly higher in number in buffaloes than in cows. Tissue samples of reproductive tracts were processed for histopathology and histopathological lesions were observed under microscope at 4X, 10X and 40X. Histopathological lesions in endometritis were hemorrhages in stratum compactum, very thick blood vessels, congestion, necrosis and degeneration of endometrial glands, glandular tissue fully filled with mononuclear cells, completely destroyed secretary epithelial cells. While in metritis, intact endometrial mucosa, proliferation of connective tissue, infilteration of inflammatory cells and mononuclear cells in the myometrium were observed. In pyometra samples, thick layer of exudate on the surface of endometrium, severe infilteration of mononuclear cells, sloughing of endometrial mucosa, necrosis of endometrial glands and glands filled with heavy population of mononuclear cells, proliferation of fibrous connective tissues in between endometrial glands, many macrophages are also present, infiltration of polymorphs mononuclear cells were observed in stratum functionalis. In uterine lymphoma, tumor tissue was in follicular pattern, neoplastic cells variable in size and having round nucleus, many mitotic figures present, and stroma consisted of broad irregular bands of collagen. In conclusion, out of 100 reproductive tracts of slaughtered cattle and buffaloes were examined and gross lesions were found 54% including uterine hemorrhages 20% followed by pyometra, para-ovarian cysts, hydrosalpinx, luteal cysts, follicular cysts and uterine lymphoma as 17, 8, 3, 3, 2 and 1%, respectively. The bacteria isolated from reproductive tracts in cattle and buffaloes were Escherichia coli 100% followed by Staphylococcus aureus, Micrococcus species, Staphylococcus epidermidis, Lactobacillus species, Corynebacterium species and Aeromonas species as 51, 47, 40, 34, 24 and 5%, respectively. In endometritis, PMN cells, thickening of endometrium, congestion, very thick blood vessels, degeneration of endometrial glands, glandular tissue fully filled with mononuclear cells, completely destroyed secretary epithelial cells, intact endometrial mucosa, proliferation of connective tissue and hemorrhages are observed while in pyometra, sloughing of endometrial mucosa, glands filled with heavy population of mononuclear cells, proliferation of fibrous connective tissue in between endometrial glands, many macrophages are also present, congestion and degeneration of endometrial glands was seen. Tumor tissue was in follicular pattern, neoplastic cells variable in size and having round nucleus, many mitotic figures present, and stroma consisted of broad irregular bands of collagen. Availability: Items available for loan: UVAS Library [Call number: 1541,T] (1).

3. Amelioration Of Pathological Effects Of New Castle Disease By Aloe Vera

by Sayyed Raza Ali Shahid (2014-VA-515) | Prof. Dr. Zafar Iqbal Chaudhry | Dr. Gulbeena Saleem | Prof. Dr. Aftab Ahmed Anjum.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Poultry industry has many threats from the infectious diseases. Newcastle disease is the most important disease of the poultry as it is distributed all over the world and it can cause huge economic losses in poultry industry. It is caused by the Newcastle disease virus (NDV) that can infect about 240 species of birds. Newcastle disease cause immune suppression in birds. It is reported that supplementations of Aloe vera enhances the immune status and reduce inflammation. So this research project was design to observe the effect of Aloe vera on lymphoid organs, growth performance and antibody response in Newcastle disease challenge birds. For this research a total of 120 broiler chicks were divided into four groups A, B, C, and D. Group A was control group while B and C were treated with 2 percent aqueous extract of Aloe vera. Group C was also vaccinated against New Castle disease. Aloe vera was given to group B and C from day one to end of study trial. Both of the groups were challenged with ND virus at day 21. Group D was vaccinated against ND and was challenged with ND virus at day 21 without supplementation of Aloe vera. Blood samples were collected at day 1, 7, 20, 24, 26 and 28 to determine the antibody titer against ND. Highest antibody titers were observed in group C as compared to all other groups which was vaccinated against ND along with supplemented with Aloe vera. For gross pathology and histopathology, lymphoid organs were collected at day 24, 26 and 28..The average feed intake of group A and D was significantly higher than group B and C before challenge of virus but the body weight gain of 2% Aloe vera supplemented broiler was significantly (p<.05) higher than without treatment of Aloe vera. The FCR of birds supplemented with Aloe vera treatment was significantly different from the birds without Aloe vera treatment. The FCR value of group C and B was higher than A and D. lxxv A significant difference was observed in the weight of lymphoid organs of birds treated with Aloe vera as weight of organs was less in group C followed by group B, group A and group D. This was due to anti-inflammatory effects of Aloe vera. Microscopic examination revealed congestion, depletion of lymphocytes, dysplasia of thymic lobules, thinning of cortex, focal necrosis, disappearance of lymph follicles and inter-follicular edema like lesions within lymphoid organ of the groups challenged with Newcastle disease virus. However, cellular hypertrophy and decreased lymphocytes population were prominent changes in lymphoid organs of broiler treated with 2% Aloe vera. To check the virucidal effects of Aloe vera, a separate experiment was conducted in which 9 day old embryonated eggs were inoculated with ND virus along with 2 percent Aloe vera gel extract after incubation at 37Cº for an hour in group A while only ND virus was inoculated in group B. Candling was performed to see the survival of embryos in both groups which revealed a significant difference i.e. 16 percent embryos were found dead in group A while 80 percent was found dead in group B. Later on the amnioallantoic fluid of the eggs was used for spot Haemagglutination test. Group A showed less agglutination activity then group B. From this study it was concluded that Newcastle disease caused immune suppression and damage of vital organs in broiler while Aloe vera have immunomodulatory, anti-inflammatory and antiviral effects as it raised antibody titer against Newcastle disease virus and lowered the inflammatory processes along with inactivation of ND virus. It also promotes growth performance of broilers and helps the birds to survive against lethal ND disease. Availability: Items available for loan: UVAS Library [Call number: 2525-T] (1).

4. Pathogenesis Of Field Isolates Of Mannheimia Hemolytica In Experimentally Infected Rabbits

by Syeda Fakhra Waheed (2014-VA-10) | Prof. Dr. Zafar Iqbal Chaudhry | Prof. Dr. Asim Aslam | Prof. Dr. Aftab Ahmed Anjum.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Shipping fever is one of the most economically important infectious diseases of ruminants with a wide prevalence throughout the continents. The disease is characterized by an acute febrile course with severe fibrinous bronchopneumonia. Infected animals may die within a few days of the onset of clinical signs, but those which survive the acute attack may become chronically infected. Both Mannheimia and Pasteurella species are commensally resident in the respiratory tract of healthy ruminants and are capable of causing infection in animals with compromised pulmonary defense system. Bovine respiratory disease (BRD) is the most common and costly problem encountered in stocker or feedlot calves. BRD also called “shipping fever”, accounts for major economic losses to the producer by reducing average daily gain, feed efficiency, and overall performance of beef calves. The aim of present study was isolation of M.haemolytica from cattle. The identification of organism was performed through biochemical tests and confirmation by polymerase chain reaction. The nature of disease was evaluated through gross and microscopic lesions. A total of 50 tissue samples (25 lungs and 25 pharynx) were collected from Punjab Agriculture and Meat company Lahore and brought to the Department of Pathology UVAS, Lahore and were analyzed for biochemical and molecular detection of M .haemolytica. For studying the pathogenesis of the disease, experimental infection was given to rabbits in Department of Pathology, UVAS Lahore. Rabbits were randomly divided into Group A, Group B and Group C with nine rabbits (n=9) in each group. Experimental infection of field isolated M. hemolytica was given intratrachealy to the rabbits. Rabbits of group A and B were infected with 0.5 mL bacterial inoculum having 103 and 106 CFU/mL respectively. The rabbits of Group C served as control group. Rectal temperature of each rabbit was recorded daily. On postmortem, CHAPTER 6 SUMMARY Summary 67 gross and microscopic lesions were recorded. The results showed that rabbits of control group not showed any gross or microscopic change. There was significant increase in rectal temperature of infected rabbits as compared to uninfected rabbits. The gross lesions were specific for the organism which was prominently observed in lungs of rabbits. The microscopic lesions revealed that there was severe consolidation, congestion and fibrin exudation in lungs of rabbits of group A which were given less number of organism and they developed clear signs of disease. The rabbits of Group B showed less prominent signs compared to group A due to early death of rabbits. There were multiple hemorrhages, of varying sizes and hyalinization of myocardial cells in infected rabbits. The severity of changes was significantly more different in Group A, as compared to Group B. It can be deduced by this study that the rabbit can be used as a model for further studies exploring the pathogenesis of the disease as the lesions resemble to shipping fever caused by M. hemolytica in ruminants. The lesions, which developed, could be descending infection resulting in typical lesions of bronchopneumonia or lobular pneumonia. Availability: Items available for loan: UVAS Library [Call number: 2517-T] (1).

5. Pathogenesis Of Aflatoxin B1 In Quails Under Experimental Conditions And Detoxification By Biological And Chemical Means

by Sakhra Mahmood (2005-VA-251) | Prof. Dr. Muhammad Younus Rana | Prof. Dr. Asim Aslam | Prof. Dr. Aftab Ahmed Anjum.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Secondary metabolites of certain fungi produce toxins under favorable conditions especially while growing on different food grains. Mycotoxins are among major threats to growing poultry industry and human beings. Aflatoxins are closely related, biologically active fungal metabolites and commonly produced by Aspergillus species. A research was carried out to evaluate the ability of Aspergillus flavus for Aflatoxin B1 production using rice, wheat and maize as substrates. Lethal effects on growth performance parameters, hematological and histopathological of graded doses of aflatoxin B1 in quails under experimental conditions were observed. Effect of Aflatoxin B1 on humoral immune response to Newcastle Disease virus vaccine in quails were determined. Biological detoxification of Aflatoxin B1 by Saccharomyces servisiae was evaluated in quails. Comparative evaluations of different commercially available toxin binders were checked. All these experiments were carried out till the six weeks (42 days). Aspergillus flavus was identified on the basis of macroscopic and microscopic characteristics. Rice, wheat and maize grains was used as substrate to check the level of Aflatoxin B1 produced by inoculating an aqueous suspension of 106 spores/ml. Aflatoxin B1 checked by Thin Layer Chromatography (TLC) and quantified by High Performance Liquid Chromatography (HPLC). Quails were reared under standard management conditions in five groups (A, B, C, D and E) having sixty each. Each group was further divided in two independent units. Diets offered to groups were control (without toxins), 0.25, 0.50, 1 and 2 mg Aflatoxin B1/kg feed. One unit of SUMMARY 187 each group was vaccinated with Newcastle Disease Virus (NDV) vaccine while other was not and studied the lethal effects on growth performance, blood parameters, immune response and histopathology of vital organs. At the end of the experiment, it was found that the deleterious effects of Aflatoxin B1 were dose and duration dependent. As the level of the toxin was increased, the lethal effects were prominent. The growth performance parameters including gain in body weight, feed intake and feed conversion ratio was adversely affected at high doses. The body weight gain was significantly reduced in Aflatoxin B1 treated groups as compared to control group. Similarly feed intake and feed conversion ratio were significantly different from the control group. The hematological studies exhibited that aflatoxin B1 significantly reduced the hemoglobin, packed cell volume and total leukocyte count whereas the erythrocyte sedimentation rate was significantly increased as compared to control group. The immune response against NDV vaccine was adversely effected in Aflatoxin B1 treated groups and values of Antibody titer in AFB1 were significantly low as compared to group A( control) In the second experiment, Saccharomyces cervisae (SC) dried powder was mixed in basal quail diet having 0.5mg Aflatoxin B1 for all experimental groups and control was without toxins. SC was added at levels of 0.5 gm, 1.0 gm and 2.0 gm /kg of feed. It was recorded that Saccharomyces cervisae (yeast) have the potential to remove the deleterious effects of Aflatoxin B1. Yeast effectively detoxified the Aflatoxin B1. The results recorded of growth performance and other parameters were non-significantly different from the control group. Chemical detoxification of Aflatoxin B1 was evaluated in quails using commercially available toxin binders. Toxin binders used were activated charcoal, kaoline, Myco AD and selenium plus vitamin E and mixed in basal quail diet having 0.5mg Aflatoxin B1 for all experimental groups and control was without toxins. The Myco AD and selenium plus vitamin E showed the highest detoxification potential as compared SUMMARY 188 to other chemical toxin binders. Groups E and F showed the results of growth performance, hematological, immune response and histopathological were non-significantly different from the control group (A). Kaolin was moderately detoxifying the toxin. Presence of aflatoxin B1 in soft tissues was checked by TLC and quantified using HPLC. The liver exhibited the residues of Aflatoxin B1 at high doses of toxin. Group D and E rearing on feeds having 1mg AFB1 /Kg feed and 2mg AFB1 /Kg feed of toxin showed the residues of AFB1 in liver and kidney. Statistical means for growth performance parameters, hematological, immune response and histopathological scores in each subunit of quails were analyzed by applying one way ANOVA and Duncans‟s Multiple Range (DMR) test at 95% probability. Aflatoxin B1 is lethal and lowers the performance of birds. The lethal effects can be detoxified by biological and chemical means to lower the economic losses to poultry industry. It can be concluded that biological detoxification is preferably better as compared to chemical detoxification. Availability: Items available for loan: UVAS Library [Call number: 2670-T] (1).

6. Identification And Molecular Characterization Of Mycobacterium Bovis And Mycobacterium Paratuberculosis In Wild Cats.

by Zianab Tariq (2010-VA-234) | Dr. Muhammad Yasin Tipu | Prof. Dr. Zafar Iqbal Chaudhary | Prof. Dr. Aftab Ahmed Anjum.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Cases of wildlife diseased with Mycobacterium species are existing in Pakistan and result in high morbidity and mortality. Vaccine is the only preventive measure, but in wildlife the vaccine administration is a strenuous job. In Pakistan vaccination practice is not up to the mark and vaccination schedules are not being followed. Mycobacterial diseases have gain popularity due to their zoonotic effect. Scat samples from Lahore Safari Zoo and Lahore Zoo were collected and properly labelled. Conventional PCR along with Touchdown PCR was done using universal primer sets of M. bovis and M. paratuberculosis. The amplicons were run on agarose gel and the bands were observed under Gel Doc system. The objective of the study was to detect the currently prevailing Mycobacterium bovis and mycobacterium avium subspecies paratuberculosis in wild cats in Pakistan. However the results obtained from different kinds of PCR were negative, showing that the wild cat population of Lahore Zoo Safari as well as Lahore Zoo were free from Mycobacterium bovis and mycobacterium avium subspecies paratuberculosis. Availability: Items available for loan: UVAS Library [Call number: 2845-T] (1).

7. Pathological Effects Of Natural And Experimental Lead (Pb) Toxicity In Lohi Sheep At Jhang, Pakistan

by Muhammad Sajid (2010VA-61) | Prof. Dr. Muhammad Younus | Dr. Muti ur Rehman Khan | Prof. Dr. Aftab Ahmed Anjum.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Heavy metal toxicity is increasing due to increasing trends of urbanization and industrialization. Lead poisoning has been recognized as a major public health risk, particularly in developing countries. It is nephrotoxic, hepatotoxic, neurotoxic, carcinogenic and mutagenic for animals and human. Sewerage water, fertilizers, leaded-gasoline and lead based batteries are the sources of lead contamination in soil and forage. The lead particles are taken up by animals from contaminated forages and excreted in animal products like milk and meat. The presence of Pb in drinking water, waste water, plant products and animal products has been studied which is a serious risk for animal and public health. The legislations for the disposal of household wastes and industrial effluents are very poor in Pakistan. The calculation of safe Pb levels in different products is still to be needed. Pathological effects of higher Pb levels have not been studied in Pakistan. The present study was aimed to unveil the toxic effects at constant dose of Pb over a period of three months in a local sheep breed of Pakistan. The status of Pb toxicity was also investigated in a polluted area around sewage drain and mutton slaughter house at District Jhang, Pakistan. The Pb concentration in soil, forage and irrigating water was found to be below the permissible limits and was safe for agriculture but long-term ingestion of low Pb concentration may have cumulative effect. The serum Pb concentration was found to be above the recommended safe limits for producing Pb toxicity in animals. The different tissues like kidney, liver and skeletal muscles also contained higher Pb level from the permissible limits and found to be unsafe for public use. Kidney showed the highest Pb concentration and the muscle contained the least Pb level in the present study. Summary 142 The erythrocyte count, hemoglobin concentration and packed cell volume showed inverse correlation with Pb concentration and mean values were below the normal range in Pb treated sheep but anemia was not developed. The erythrocyte sedimentation rate was also influenced by given dose of lead acetate during third month of treatment. The white blood cells also revealed no effect on given dose of lead acetate in Lohi sheep in this study. The biochemical parameters of field and treatment group showed higher concentration as compared to control group of Lohi sheep but their means were falling within the normal range of reference values. The disturbed biochemical parameters in apparently healthy sheep with higher serum Pb concentration were indicative for liver and kidney damage. Lohi sheep exhibited less effect on given dose of lead acetate during first two months but more pronounced changes of chronic Pb toxicity were observed during last month of trial. The histological changes were not observed on early period in lead acetated treated sheep. The characteristic histological changes were observed on last slaughtering at day 90 in kidney and liver including degeneration and focal areas of necrosis, dilatation of blood vessels with accumulation of red blood cells and fibrosis in some areas. The nuclear changes were more typical with intranuclear inclusion bodies in renal tubular epithelial cells but less distinguishable in hepatocytes. It was concluded that soil, forage and water contained low Pb levels in the study area. The ingestion of low Pb level for longer period had cumulative effect in animals. The animals might be resistant to low Pb level but their products are a severe risk for public health. So the necessary measures should be adopted to minimize the heavy metal contamination in animal products. Availability: Items available for loan: UVAS Library [Call number: 2820-T] (1).

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