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1. Molecular Characterization Of Brucella Abortus Strains In Bovines

by Muhammad Ramzan | Dr. Raheela Akhtar | Prof. Dr. Aneela | Prof. Dr. Asim Aslam.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2169,T] (1).

2. Pathological Studies On Contagious Edthyma In Naturally Infected Small Ruminants

by Muhammad Usman ghani | Dr. Mati ur rehman khan | Dr. Muhammad | Prof. Dr. Asim aslam.

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2070,T] (1).

3. Detection Of Influenza A Virus Contamination In Newcastle Disease Live Virus Vaccines And Their Pathological Effects On Visceral Organs

by Munir Hussain (2004-VA-64) | Mr. Muhammad Saeed Imran | Prof. Dr. Asim Aslam | Dr. Shafqat Fatima Rehmani.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Poultry is one of the most vibrant commercial sector which is playing a vital role to bridge the gap between supply and demand of animal protein foods to cater for its ever increasing human population 2.1 per cent annually in Pakistan (Sahota et al. 2003). Vaccination is one of the most effective way to prevent the poultry birds from the specific diseases. Disease producing microorganisms can be classified smallest to largest as viruses, bacteria, fungi, protozoa and parasites. All, except the viruses are sensitive to drugs when outbreaks occur. Vaccination is basically the introduction of a specific biological substance (antigen) into the bird to stimulate the antibodies formation or immunity to a particular disease. Usually the biological substance is avirulent the live disease organisms, which are capable to protect the bird against the particular disease by producing an immune response. Presence of these organisms (antigen) in the blood stimulates the body's defense mechanism to produce antibodies that neutralize the disease causing organisms when the bird is exposed to them (Kamboh et al. 2009). A danger of such type of live vaccines is that the live microbes can back mutate to a virulent form. While, dead vaccines that contain whole killed (usually by formalin or phenol) microbes are safe. They may contain little or no extraneous material and therefore tend to produce fewer adverse effects (Palombo and Semple 2001). The vaccines that contain dead organisms are safe with respect to residual virulence and are easy to store, since organisms are already dead. While live vaccines may possess residual virulence for the animal by reversion of avirulent organisms to fully virulent type or spread to nonvaccinated animals. Dead vaccines have very little risk of ‘alive’ contamination, while live vaccines always run the risk of contamination with unwanted organisms; for instance, outbreaks of reticuloendotheliosis in Introduction ______________________________________________________________________________ 2 chickens in Japan and Australia have been traced to contaminated Marek’s disease vaccine (Tizard 1995). Avian Influenza viruses typically produce Syndromes ranging from asymptomatic infection to respiratory disease and drops in egg production to severe, systemic disease with near 100% mortality (Olsen et al. 2002). Avian influenza initially was recognized as a highly lethal, systemic disease (i.e., highly pathogenic). HPAI was known by various name including fowl plague, fowl pest etc. Avian Influenza viruses are classified in the family orthomyxoviridae, genus influenza virus A (Garten et al. 2009). Avian influenza viruses can be categorized into four clinical groups:1) highly virulent, 2) moderately virulent, 3) mildly virulent, and 4) Avirulent (Swayne and Suarez 2000). Avian Influenza further sub type based on serologic reaction of HA and NA surface glycoproteins. Fifteen sub types of HA and nine sub types of NA are recognized (Swayne and Suarez 2000). MP AI viruses in domestic poultry produce clinical sign reflect abnormalities in the respiratory, digestive, urinary and reproductive organs (Allwright et al. 1993). To date, naturally occurring highly virulent influenza A viruses that produce acute clinical disease in chickens, turkeys and other birds of economic importance have been associated only with the H5, H7 and H9 subtypes. Influenza A viruses of subtype H9 are now considered to be wide spread in poultry and have demonstrated the ability to infect humans (Fedorko et al. 2006). To date, all outbreaks of the highly pathogenic form have been caused by influenza A viruses of the subtypes H5 and H7. The disease is transmitted horizontally by direct contact through contamination. There is little or no evidence of vertical transmission (egg-borne infection). However, eggshell surfaces can be contaminated with the virus (Potima 2007). Wild and domesticated water fowl is the major natural reservoir of influenza A viruses. Representatives of Introduction ______________________________________________________________________________ 3 all of the different subtypes of avian influenza A virus have been isolated from birds, particularly from aquatic species such as ducks, geese, and gulls (Karasin et al. 2000). Wild birds such as geese, ducks and game birds; they can be carriers of even highly pathogenic strain H5N1 shedding the virus in their feces without clinical signs of disease. Thus, the present study was carried out to examine the viral contamination (Influenza A virus) in poultry vaccines manufactured locally and imported from different countries of the world in Pakistan. The findings of the study have helped us to see the Avian Influenza A virus contamination in vaccines which are used in field conditions and also help to evaluate the purity of vaccines. The RT-PCR based technology has been described for the detection of different RNA viruses such as Newcastle disease virus etc. (Payne et al. 1981) revealed contamination of vaccines with ALVs, specifically in two Marek´s vaccines, which confirms that these agents are potential contaminants of viral vaccines applied in poultry. This assay has meant a considerable advance due to a higher sensitivity and specificity upon differentiating the subgroups compared with ELISA. It is quicker test for detection of RNA viruses than the viral isolation, which requires until 10 days and it needs detection by ELISA for the identification result. Availability: Items available for loan: UVAS Library [Call number: 2212,T] (1).

4. Effect Of Temperature And Relative Humidity On The Survival Of Newcastle Disease Virus Isolates Using Germ Carrier Techniques

by Tayyeba Sohail (2009-VA-209) | Dr. Jawad Nazir | Prof. Dr. Khushi Muhammad) | Prof. Dr. Asim Aslam.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Newcastle Disease (ND) is a highly contagious viral disease that affects almost all avian species including poultry, cage and wild birds around the globe (Terregino et al. 2003; Vidanovic et al. 2011). ND is economically important disease and included as list-A disease of Office des International Epizootics (OIE) (Anonymous). Mortality of infected birds ranges from negligible to as high as 100 % depending on the pathotype of the virus involved and health status of the birds (Alexander and Manvell 2004). NDV is an enveloped virus with single stranded, non-segmented, negative sense RNA genome (Makoui et al. 2013). The virus belongs to Avulavirus genus of Paramyxoviridae. There exist only one serotype of NDV designated as avian paramyxovirus-1 (Kapczynski et al. 2013) however, different virus strains do vary in their pathogenicity. There are 3 pathotypes of NDV; velogenic (highly virulent), mesogenic (moderate virulent), and lentogenic (mild virulent) based upon diseases producing potential and severity of signs in the infected birds (de Leeuw and Peeters 1999). NDV is primarily transmitted to the susceptible birds through aerosol and fecal oral route (Martin 1992). Infected birds secrete high amount of the virus in their feces, saliva, mucous and nasal secretions which might contaminate the premises. Inanimate objects or fomites are a potential reservoir of viruses outside the host and might play an important role in the transmission of pathogens (Nicas and Sun 2006). Several factors can influence the survival of viruses outside the host (Sobsey and Meschke 2003; Weber and Stilianakis 2008; Stallknecht and Brown 2009). A number of studies show that respiratory pathogens can survive from hours to months on fomites (Abad et al. 2001; Kramer et al. 2006). Certain physical factors like temperature, humidity, pH, salinity, exposure to ultraviolet (UV) rays etc drastically affects the Introduction 2 virus persistence in the environment. Effect of such physical insults is more pronounced on enveloped viruses than non-enveloped ones (Mbithi et al. 1991; Schaap et al. 2012; Tuladhar et al. 2012). High humidity and temperatures not only reduces the survival of influenza viruses on contaminated surfaces but also modulates their transmission to the susceptible birds (Shaman and Kohn 2009; McDevitt et al. 2010; Paynter 2014). Similarly lower temperature and less humidity promote the survival of NDV in the environment (Dat and Chuc 1985; Kournikakis et al. 1988). ND is endemic in Pakistan but since last few years several new virus strains are circulating in commercial and rural poultry of the country (Munir et al. 2012; Shabbir et al. 2013). Central Punjab region is densely populated with commercial poultry and serve as disease epicenter every year. It has been observed that the disease outbreaks usually start in December, attain peak in the late winter and spring season, start decline in June and disappear in the rainy season. Apart from several other contributing factors, environmental survival of the viruses might contribute to the disease outbreaks. Availability: Items available for loan: UVAS Library [Call number: 2227-T] (1).

5. Effect of Curcuma Longa on Embryonated Eggs Experimentally Infected With Avian Influenza Virus

by Syed Iftikhar Ali Shah (2013-VA-436) | Dr. Muhammad Yasin Tipu | Prof. Dr. Asim Aslam | Prof.Dr.Muhammad Sarwar Khan.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: CD Error Availability: Items available for loan: UVAS Library [Call number: 2264-T] (1).

6. In Process Quality Control Factors Affecting The Quality Of Locally Prepared Salmonella Gallinarum Antigen

by Zahra Malik (2009-VA-245) | Dr. Arfan Ahmad | Prof. Dr. Aftab Ahmad Anjum | Prof. Dr. Asim Aslam.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Fowl typhoid is a septicaemic disease caused by S. gallinarum biovar gallinarum has major economic significance in many parts of the world. It is an acute or chronic septicaemic disease that usually affects the birds (mostly adult birds). Eradication of disease is normally done by identifying the infected flocks and eliminating the reactor birds by using serological tests, but diagnosis of the disease is much expensive because antigen used for this purpose is imported. The study, therefore, has been proposed to prepare and evaluate the stained antigen of S. gallinarum using local isolates. A total of 15 isolates were procured from Poultry Research Institute (PRI) Rawalpindi, University Diagnostic Lab (UDL) and Department of Microbiology, UVAS Lahore, which were identified by Biochemical testing and further confirmed by Polymerase Chain Reaction. Among all 15 isolates two isolates were confirmed as S. gallinarum and proceeded to prepare local antigen of S. gallinarum. Locally prepared antigen was checked with known positive and negative sera, Effect of different preservatives (Sodium azide and Thiomersal sodium) and different storage temperatures (4°C, 25°C and -20°C) was also studied after every fifteen days post storage upto 6 months to observe the stability and shelf life of local antigen. On the end of study both preservatives i.e. Sodium azide and Thiomersal sodium was found equally effective for antigen activity, whereas 4°C proved best storage temperature to be used for the antigen preservation. Activity of locally prepared antigens was also compared with the imported antigen (Charles, River, USA) stored at different temperatures regularly throughout the six months, which showed that local antigens was almost as good as the imported antigen. Summary 51 CONCLUSION Locally prepared S. gallinarum antigen was found as effective as imported antigen. Both the test preservatives (Sodium azide and Thiomersal Sodium) had the same effect on antigen preservation. Among all three test temperatures, 4°C was accepted as best storage temperature for the long term preservation of local antigen with either of the preservative. Availability: Items available for loan: UVAS Library [Call number: 2278-T] (1).

7. Pathobiological Studies On Bovine Ephemeral Fever Infected Cattle In District Swabi

by Sahibzada Waheed Abdullah (2013-VA-560) | Dr. Muti Ur Rehman Khan | Prof. Dr. Asim Aslam | Dr. Ali Ahmad Shiekh.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Among the non-contagious diseases Bovine ephemeral fever is important disease of cattle the course of the disease is usually three days due to which it is called “three days sickness”. This is transfer to other cattle through insects Culicoides (biting midges a group that include many kinds of flies) and mosquitoes. Bovine ephemeral fever virus (BEFV) has been collected from Culicoides coarctatus, Culicoides brevitarsis, and Anopheles bancroftii (Walker et al. 2012). Cattle and buffaloes are the main species affected from Bovine ephemeral fever (BEF) which gives huge economic losses to the dairy sector. The etiologic agent, Bovine ephemeral fever virus belong to Rhabdoviridae family, enveloped (negative sense) ssRNA virus. It generally recur in Australia, Asia, and Africa also in the Middle East (Walker 2005). The theme of the present study was detection of BEF virus through Reverse Transcriptase-Polymerase Chain Reaction from the cattle suspected for bovine ephemeral fever virus on the basis of clinical signs. Hematological profile and serum calcium level were checked in the confirmed positive samples for BEFV. A total of 50 blood samples were collected from the suspected animals in aseptic condition using a sterilized disposable syringe and were preserved in vacutainers (Anticoagulant added n = 50, without anticoagulant n = 50). The 10 blood samples were collected from healthy animals in vacutainers (Anticoagulant added n = 10, without anticoagulant n = 10). Buffy coat were separated from blood samples and from this the RT-PCR was performed and successfully diagnosed the BEFV infected cattle. Hematology and serum calcium were performed for both confirm positive and healthy animals. Summary 31 The result showed that BEF virus was diagnosed with the help of RT-PCR in samples suspected for BEFV infection, and there was no virus detected in samples taken from healthy animals. Comparison of hematology between BEFV infected cattle and healthy animals were performed there was no changes in the RBC, Hemoglobin, Hematocrit, MCV, MCH, MCHC, and MID (it include monocyte, eosinophils, and basophils) except Neutrophils, which number was increases and lymphocytes which was decreased in BEFV infection, while in healthy animals there was no change in the whole hematology. Serum calcium was also determined there was decrease in serum calcium level of BEFV infected cattle, while in the healthy animal samples there was no change in the serum calcium level. Availability: Items available for loan: UVAS Library [Call number: 2284-T] (1).

8. Effect Of Citrullus Colocynthis On Serobiological Parameters In Alloxan Induced Diabetic Rats

by Farah Javed (2012-VA-398) | Dr. Muhammad Quaid Zaman | Dr. Imtiaz Rabbani | Prof. Dr. Asim Aslam .

Material type: book Book; Format: print ; Literary form: not fiction Publisher: 2015Dissertation note: Diabetes mellitus is one of the most common endocrine disorders affecting almost 25% of the world's population.The pretentious worldwide increase in the incidence of diabetes mellitus is posturing a huge health problem in both developed and developing countries. Diabetes mellitus is a metabolic disorder which is manifested by polyuria, polyphagia, polydipsia, hyperglycemia and dyslipidemia and is still one of the most leading causes of disability and death. Oral hypoglycemic agent and insulin are common treatment of diabetes but these treatments have prominent side effects. In the recent years the use ofherbal medicines has increased for the treatment of diabetes and fascinated the consideration ofmany researchers all over the world. Citrullus colocynthis is used commonly in different parts of the world for the treatment of a number of diseases including diabetes, jaundice, leprosy, cancer, asthma, bronchitis, joint pain and mastitis. In the present study I evaluate the anti-diabetic effects of roots of Citrullus colocynthis and also the dose dependent anti-diabetic effects of medicine in alloxan induced diabetic rats. Twenty-five adult male rats were divided into five groups; Negative control, Positive control, and 3 groups for different treatment dose of roots of Citrulllus colocynthis (200 mg/kg body weight, BW), Citrulllus colocynthis (300 mg/kg BW) and Citrulllus colocynthis (500 mg/kg BW). Diabetes induction had done in four groups, other than negative control (normal saline injected), by subcutaneous administration of alloxan (120 mg/kg BW). Blood glucose level of rats reached above 250mg/dl considered as hyperglycemic. Treatment was given to all groups excluding control negativefor 21 days. Body weight of rats of all groups was recorded weekly. After completing 21 days of treatment with different doses of roots of Citrullus colocynthis blood samples were collected in fasting condition from rats of each group by cardiac punctureunder general anesthesia. Serum was collected from blood to measure serum glucose level, serum lipid profile, liver function test and renal function test. Result data was analyzed by using SPSS software. Data was analyzed by using one-way ANOVA. The group differences were compared by the Duncan’s Multiple Range Test. Differences was considered significant at P < 0.05. The obtained results showed that roots of Citrullus colocynthis has efficiency to control the diabetes mellitus by reducing serum glucose levels as well as the increasing dose decreased the serum glucose levels. Only 500mg/kg body weight dose is efficient to reduce the muscle wastage due to diabetes in alloxan induce diabetic rats. This dose also works tomaintain the serum ALT, AST, urea, creatinine, total cholesterol and triglycerides, HDL-C LDL-C levels. The data obtained from this study also show the dose dependent anti-diabetic activity of medicine as the dose of 500mg/kg body weight is more effective to control the diabetes as compared to other two doses; 200mg/kg body weight and 300mg/kg body weight. Availability: Items available for loan: UVAS Library [Call number: 2297-T] (1).

9. Effect of Fish Oil on Response of Lymphoid Organs of Broiler Experimentally Infected With Newcastle Disease Virus

by Muhammad Zahid (2013-VA-441) | Dr. Muhammad Yasin Tipu | Dr. Muhammad Yasin Tipu | Prof. Dr. Asim Aslam | Prof. Dr. Khushi Muhammad.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Theses submitted with cd. Availability: Items available for loan: UVAS Library [Call number: 2353-T] (1).

10. Distribution and Localization of Brucella Melitensis in Aborted Fetal Tissues of Small Ruminants

by Muhammad Zain Saleem (2008-va-158) | Dr. Raheela Akhtar | Prof. Dr. Asim Aslam | Dr. Haroon Akbar.

Material type: book Book; Literary form: not fiction Publisher: 2015Dissertation note: Submitted with blank CD. Availability: Items available for loan: UVAS Library [Call number: 2369-T] (1).

11. Pathological Investigations Of Different Isolates Of H9n2 Prevalent In Broiler Chicken

by M. Furqan Shahid (2014-VA-322) | Dr. Yasin Tipu | Prof. Dr. Asim Aslam | Prof. Dr. Tahir Yaqub.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: In recent years, H9N2 virus has attained a great importance as its infection has reached panzootic proportions. AIV H9 has different antigenic variants that has made it problematic to diagnose and thus to understand the pathogenicity of this virus is also very difficult. Detection of AI H9 antibodies can be used as a complementary method for sero-epidemiological studies as an indicator of AI H9 infection. The haemagglutination inhibition (HI) assay is used routinely for subtyping and detecting an increase of antibodies to AI viruses. Surveillance and early diagnosis of AI virus is essential for poultry. It demands rapid, sensitive and inexpensive diagnostic tests. Thus, it is important to identify different antigenic variants of H9. In this study a total of seven H9 virus samples were isolated out of total 100 collected sample from field outbreaks. These isolates were confirmed by molecular methods like PCR. Then four isolates from these seven isolates were used to infect the experimental broiler chicken. Clinical signs were recorded after the inoculation of H9N2 virus to the broiler. The results of this study showed that clinical signs were more sever upto 5 DPI. The severity of signs was proved by observing the gross pathology and histopathology of organs (Lung, Kidney, Trachea and Liver) of infected birds which were collected on 5 and 9 DPI. Serum of infected birds was also collected on 7 and 14 DPI to analyze the antibody level of infected birds against experimentally used isolate of H9N2. Then cross reactivity of different isolates of H9N2 was also checked against pannel of hyperimmune sera raised against different isolates of H9N2 and their results showed different antibody level against different isolates of H9N2. The sero-biochemical study of serum of infected birds revealed that H9N2 virus has pathogenic potential on kidney and liver. Availability: Items available for loan: UVAS Library [Call number: 2459-T] (1).

12. 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).

13. Effect Of Acetic Acid Supplementation On Pathomorphological And Immunohistochemical Changes In Broiler Chickens Experimentally Infected With Salmonella Enterica Serovar Pullorum

by Bareera Javed Khan (2009-VA-156) | Dr. Gulbeena Saleem | Prof. Dr. Asim Aslam | Dr. Nisar Ahmed.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: The present study was conducted to evaluate the effects of acetic acid in minimizing the severity of pathomorpholgical lesions in broiler chickens experimentally challenged with Salmonella pullorum. The experimental birds were divided into five groups. Group A acted as control, Group B was infected with S. pullorum. Antibiotic and acetic acid was given respectively to the challenged Group C and Group D. Group E was given acetic acid solely. Clinical signs were observed on daily basis. Postmortem findings of birds from each group was recorded on day 1, 3, 5 and 7. Histopathology and immunohistochemistry of the necropsy samples was performed subsequently. The data thus collected was organized using Factorial experiment on computer statistical software Minitab version 16 and analyzed by Two way ANOVA (Analysis of variance). Hemorrhagic, congested liver with greyish necrotic foci, pericarditis, congested lungs, spleen and unabsorbed yolk was observed in sick birds. Infiltration of inflammatory cells, congestion and necrosis in liver, spleen and heart were histopathologically observed. Acetic acid reduced the severity of gross pathological and histopathological changes. The fecal excretion of S. pullorum significantly reduced with acetic acid. Results clearly demonstrated that use of acetic acid and antibiotic respectively produced comparable outcome. As the use of antibiotics was banned in European Union and the organism, Salmonella pullorum showed resistance against many antibiotics so the best way to control the disease is by supplementing the acetic acid to birds as it was helpful in minimizing the mortality and severity of gross and histopathological lesions in infected chickens. If diets can be planned to enhance the organic acid production in the caecum, it may be possible to control salmonella species through cost effective means. However further studies need to be conducted in order to analyze the prophylactic and therapeutic effect of organic acids. The use of prebiotics and probiotics along with organic acids on the growth and disease management of broiler chickens. Availability: Items available for loan: UVAS Library [Call number: 2564-T] (1).

14. Isolation And Molecular Characterization Of Rotavirus From Calf Diarrhea And Preparation Of Vaccine

by Nadia Mukhtar (2008-VA-718) | Prof. Dr. Tahir Yaqub | Dr. Jawad Nazir | Prof. Dr. Asim Aslam.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: The main contribution of the thesis “Title” is threefold. First, rotavirus was isolated and identified from calf diarrhea samples from 10 districts in Punjab. Second, optimization of molecular diagnostics and genome sequencing was done of the positive bovine rotavirus isolates from Pakistan. And thirdly, the preparation as well as evaluation of killed vaccine against bovine rotavirus isolates was performed. The above three objectives of this study were created due to the distribution of rotavirus all over the world as an enteric pathogen in both human as well as animal species. In developing countries where cases of malnutrition are very common in young children and animals, this virus has a special importance as an etiologic agent. It causes severe diarrhea, when accompanied with severe dehydration, leads to high rate of mortality. Among the rest of the infectious diseases present in calves, neonatal diarrhea is a dire threat as it has a major impact on economic viability. Calf diarrhea is the most important problem in dairy calves that causes more financial losses to the calf producers than any other. Although numerous etiological agents may be implicated, Rotaviral diarrhea is one of the main infections causing calves to scour between five to fourteen days of age. The cattle and buffalo calves’ population in Pakistan is devastatingly affected by the neonatal calf diarrhea due to rotavirus outbreaks. Neonatal calf mortality varies from 8.7 to 64 per cent throughout the world accounting for 84 per cent of the total mortality in the first month of age and is particularly high in the third week. While vaccination is available for the disease, it is being imported in Pakistan from other countries. The importation of the said vaccine thus, leads SUMMARY 117 to extra expenses for the farm managers. As mentioned above one of the aims of this study is to develop an effective vaccine against bovine rotavirus and cut down expenses for farm managers. To fulfill the objectives proposed in this thesis, rectal swabs and fecal samples were collected from public/private sector buffalo and cattle farms from 10 districts of the Punjab: Lahore, Faisalabad, Okara, Sahiwal, Sargodha, Chakwal, Bhakkar, Bahawalnagar, Multan and Bahawalpur. The samples were selected on the basis of agro-ecological zones of the province. As sampling based on agro-ecological zones allow for better data collection for recording incidence rate of the disease. Samples (n=10) from each diarrheic and apparently healthy cattle and buffalo calves from all of the districts were collected. In this way a total of 200 samples from buffalo calves and 200 samples from cattle calves were collected for this study. Antigen of bovine rotavirus was screened from calf feces through Direct Sandwich ELISA. Bovine rotavirus samples were further confirmed through the amplification of the VP4 and VP6 genes through Rt-PCR. Homology and phylogenetic analysis of the sequenced samples was also performed. The data gathered through this analysis was helpful in collecting important data regarding the similarities as well as differences of the bovine rotavirus strain present in Pakistani isolates when compared to local regions as well as international ones. The data is also valuable when it comes to production of effective vaccines again rotavirus. RNA viruses are known to mutate unpredictably and it is safe to assume that a particular vaccine might not work effectively against all strains of a particular virus. That’s why analysis of data pertaining to all possible BRV strains is important for creation of an effective vaccine of import quality in order to help the economy of Pakistan. Rotavirus isolate, after adaptation on MDBK cell line, was further propagated to determine TCID50 for vaccine preparation purposes. Final dose of the vaccine was adjusted to SUMMARY 118 approximately 3ml, containing 40% culture and 60% adjuvant. Final vaccine contained 1ml of inactivated bovine rotavirus harvested culture, 1.8ml of Montanide ISA 70, 0.2ml of PBS and 0.05% of Thiomersal sodium. Efficacy of the vaccine was checked in rabbits. For vaccine efficacy testing twenty one month old rabbits were procured. Rabbits were reared in individual isolator units in the shed facility of Quality Operations Laboratory, UVAS, Lahore. The collected rabbits were divided into two groups, vaccinated and unvaccinated rabbit groups. Each group had 10 rabbits. One ml of rotavirus vaccine was administered intramuscularly in vaccinated rabbits group. In unvaccinated rabbits group 1ml of normal saline was injected intramuscularly. The second dose of vaccine was administered at 24 days post-vaccination of first dose. The rabbits from both groups were bled at 0, 14, 28 and 42 days post-vaccination. The antibody response of rabbits to rotavirus vaccine was determined through using Antibody detection kit. The rabbits were challenged on day 42 post-vaccination using live field strain of rotavirus having TCID50 1 × 108.5. The rabbits were observed daily up to 14 days post-vaccination for appearance of diarrheic signs. The stool samples of ELISA positive were further confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR) at least 14 days post-vaccination. The field trials were conducted at Livestock Production Research Institute, (LPRI) Bahadurnagar, Okara. The field study was done to evaluate the prepared rotavirus vaccine for prevention of neonatal calf diarrhea. For this trial, 100 dams were selected. The dams were divided into two groups and each group consisted of 25 pregnant cows and 25 pregnant buffalos. A total of 50 dams (25 cattle and 25 buffalo) were vaccinated intramuscularly with 3ml of prepared inactivated rotavirus vaccine. The 50 remaining dams (25 cattle and 25 buffalo) were kept unvaccinated. SUMMARY 119 The blood samples were collected for serum separation after 0, 14, 28 and 42 days post vaccination in dams. The antibody titers were measured using antibody detection ELSIA kit. After calving, newborn calves were fed with the colostrum obtained from the vaccinated dams daily for 5 consecutive days. Similarly, the calves from unvaccinated dams were fed on colostrum from their unvaccinated dams. The 5 calves from vaccinated and 5 from the unvaccinated dams were isolated in individual isolators. These calves were challenged orally with 1ml of live field strain of rotavirus having 1 × 108.5 TCID50 and the animals were observed for diarrheic signs for 7 days. All of the collected data was subjected to statistical analysis of (one way) ANOVA and t-test using SPSS. The <0.05 p-value determined the significance of the results through this study. The data collected through this study allowed for the creation of valuable inferences. According to the current results of this study, the prevalence of bovine rotavirus was shown to be 6% in Punjab. This 6% included 40% and 20% from the districts of Lahore and Faisalabad respectively. Keeping these results in mind, it is to be noted that the recorded prevalence percentage from this study is higher than the prevalence of 2% in Lahore according to a previous study done in the country. It is to be noted that while the 6% prevalence of rotavirus in Punjab detected through ELISA is lower than the prevalence of 16.83% which was detected by ELISA in diarrheic calves from pervious researches, the 12% prevalence detected by ELISA in this research is higher than the prevalence of 7.25% detected by ELISA in diarrheic calves from past data. In the present study of this thesis it was observed that the use of killed vaccine for bovines produced more efficient immune response in calves. It also enhanced the clostral rotavirus antibody titers as compared to previous studies where the use of the same strain of modified-live virus in a commercial vaccine administered IM with or without adjuvant did not significantly SUMMARY 120 elevate colostrum antibody titers. The results collected from the present research showed that the average antibody titers in the 25 cattle dams at 0, 14, 28 and 42 days post vaccination were 0%, 57%, 68% and 78% respectively. In a similar manner the average antibody titers in the 25 buffalo dams at 0, 14, 28 and 42 days post vaccination were 0%, 55%, 70% and 82% respectively. These results indicated the protective maternal antibody level against the rotavirus which will be transferred passively to calves. The results indicate that vaccinated dams were able to provide passive immunity to both buffalo and cattle calves in order to provide protection against the deadly virus. Availability: Items available for loan: UVAS Library [Call number: 2570-T] (1).

15. Comparative Hematology And Histopathology Of Parvo Virus And Corona Virus Infections In Dogs

by Qazi Abdul Aziz (2012-VA-984) | Mr. Irfan Irshad | Prof. Dr. Asim Aslam | Prof. Dr. Habibur Rehman.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Canine viral enteritis is a disease of dogs with an acute onset of vomiting and diarrhea, especially in puppies. Four viruses have been identified as the essential cause of severe enteritis in dogs: Canine Parvo Virus (CPV), Canine Corona Virus (CCV), Canine Rota Virus and Canine Distemper Virus (Jones et al. 1997; Buonavoglia et al. 2006). CPV is a contagious viral disease of dogs and is one of the most important causes of deaths in puppies (Decaro et al. 2005). Puppies aged between six and twenty weeks old, are most susceptible to CPV. CCV was first isolated in 1971 from gastro enteric dogs. Subsequently additional cases have been reported that were usually mild and self-limiting, unless complicated by CPV (Jones et al. 1997). CCV is mainly associated with respiratory, enteric, hepatic and central nervous system diseases. Nevertheless, organs such as kidney, heart, and eye can also be affected. Infections are usually self-limiting but may fatal in young animals (Pratelli et al. 2004; Evermann et al. 2005).Canine coronavirus (CCV) and canine parvovirus (CPV) are pathogens responsible for acute gastroenteritis in dogs (Decaro et al. 2008; Holzer and Parrish 2010). Canine coronavirus infection was regarded as a mild, self-limiting infection of the small intestine, especially in puppies (Decaro et al. 2008). CPV and CCV are immensely infectious viral diseases of dogsof all ages but young pups are mostly affected. Vulnerability of infection depends on age and immune status of animals. Infection is more severe in young dogs.A total of fifty clinically positive animals were selected with strong clue of gastro enteritis at various pet clinics in district Lahore. Gross pathological examinations of animals were done prior to sampling.Anorexia, emaciation, vomition, foul-smelling bloody diarrhea, temperature, depression, rough coat, and color of mucous membranes were gross pathological finding in positive animals. Less than six months ages of dogs were more affected as compared to dogs above from this age. Similarly small pupswere more challenging. Histopathology and hematology was done for positive animals. Hematological examination was statistically analyzed with one way ANOVA with the help of SAS version 9.1. Results were statistically significant and there was decrease in WBCs, Lymphocytes count and platelets count. Histopathological studies revealed the degeneration of intestinal epithelium, infiltration of mononuclear cells principally macrophages and neutrophils and blood vessels were filled with RBCs. Objectives of current study was to characterize the clinical, hematological and histopathological findings in dogs diagnosed with CCV and CPV natural infections in order to explore their usefulness as laboratory markers for the differential diagnosis of CPV and CCV. Availability: Items available for loan: UVAS Library [Call number: 2641-T] (1).

16. Histopathological Studies On Caprine Mastitis Correlating Lesions With Etiology In Natural Infection Prevailing In Lahore Abattoirs

by Salman Ahmed Abid (2014-VA-536) | Prof. Dr. Zafar Iqbal Chudhary | Prof. Dr. Asim Aslam | Prof. Dr. Aneela Zameer Durrani.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Mastitis is a common disease of cattle, buffaloes, dairy and non-dairy goats associated with the inflammation of mammary parenchyma, protracted production loss, risks of premature culling from the herd and the release of injurious toxins in the udder. IMIs in dairy goats can cause economic losses due to decreased milk production as well as risks to public health and discarded milk. A total of one hundred goats affected with mastitis were included in this study. Samples were collected from the abattoirs of Lahore. Mastitis was diagnosed on the basis of visible and palpable changes in udder and milk. Pre-slaughter and post slaughter examination of udder was performed and gross lesions were observed. Samples included udder parenchyma and supramammary lymph nodes from mastitis affected goats. Each sample was divided into two parts, one part was placed in small polyethene bag in an ice box under aseptic conditions for bacteriological examination and second part was fixed in 10% neutral buffered formalin solution for histopathological evaluation. Samples were cultured for identification of staphylococci, streptococci and E.coli on Staph 110, Blood agar and MacConkey’s agar respectively. Biochemical tests were also performed for confirmation of these bacteria. Confirmation was made on the pattern of reactivity of bacterial cultures to biochemical tests. Bacteriological investigation demonstrated the different species of bacteria involved commonly in caprine mastitis. Staphylococcus aureus was isolated from 21 cases, CNS from 10 cases, Streptococcus spp. from 7 cases and E.coli from 3 cases as single infection and 25 cases of mixed infection were observed in different combination of these bacteria. Results of the study Summary 47 revealed that Staphylococcus aureus is associated with statistically significant changes in udder parenchyma as well as in supramammary lymph nodes. Marked changes have been observed in case of tissue necrosis, exudation and gangrene. Moreover, tissue responses to mononuclear cell infiltration have also been observed significant in Staphylococcus aureus infection. CNS, Streptococci and E. coli revealed relatively comparable changes in tissue with slight variability. However, mixed infection of these bacteria in a single tissue led to relatively much pronounced histopathological changes as compared to the solitary infections. This could be attributed to the synergistic effects of various bacterial activities, enzymes, toxins and host responses to more than one type to bacteria. Availability: Items available for loan: UVAS Library [Call number: 2652-T] (1).

17. Hematological And Biochemical Study Of Haemoparasitism In Camel In Attock

by Tamoor Azeem (2014-VA-537) | Dr. M Yasin Tipu | Dr. Sajjad Ahmed | Prof. Dr. Asim Aslam | Dr. M Avais.

Material type: book Book; Literary form: not fiction Publisher: 2016Dissertation note: Camel is a multipurpose animal. Its role in the daily life cannot be ignored, it plays important role in food chain by adding meat, milk and milk products. Hides and hairs of camels are also used in tannery and cosmetic industry. Camels have ability to produce 15-20 litres of milk if proper management and feeding practices are established on farms. Fattening of camel calves can result in increase of 1kg of meat per day. Ability of camels to survive in both extreme hot and cold weather makes it a unique animal. Production of camel in Pakistan is at its modest level in most of the areas due to improper feeding and management. Haemoparasitism is a major setback in the production of camel. The study was carried out on camels in villages of six tehsils of district Attock under natural conditions. Blood samples were carefully collected from randomly selected camels from different villages. Wet smear was made on the spot by blood from marginal vein of ear and was fixed using ethanol then 10 ml of blood was collected from the same camel from jugular vein by using 10 ml sterile disposable syringe, 3ml blood was stored in ca-edta vacutainor and other 5 ml in plain vacutainer. Age, sex and breed of the animal were carefully noted. After the collection of sample, it was transported to government district laboratory where the serum was separated from the sample in a plain vacutainer by centrifugation and was freezed the sample was then transported to department of pathology, university of veterinary and animal sciences Lahore in cold chain. Wet blood smear was stain with field stain, and then these smears were used for haemoparasite detection. CBC and serum chemistry were done. Results showed that haemoparasitism causes alteration in haematology and serum chemistry of the sample which can be helpful for diagnosing as well as differential diagnosis of haemoparasites in camels. Availability: Items available for loan: UVAS Library [Call number: 2649-T] (1).

18. 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).

19. Effect Of Bacillus Subtilis And Sodium Butyrate On The Morphometry Of The Small Intestine And Immune System In Healthy And Salmonella-Challenged Broiler Chickens

by Arbab Sikandar (2005-VA-154) | Dr. Hafsa Zaneb | Prof. Dr. muhammad Younus | Dr. Sima Masood | Prof. Dr. Asim Aslam.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Supplementation ofBacillus subtilis and microencapsulated sodium butyrate in the feed is being practiced as a substitute for antibiotics growth promoters. An expansive range of encouraging health-related properties exhibited by B. subtilis and SB has been published, but their exact effect on gut and immune system is not completely understood. Consequently, the evaluation of B. subtilis andSB as feed supplements is desired. To achieve this goal, the present study was aimed to investigate the effects of B. subtilis and SB on performance, immune system, gut and lymphoid organs microarchitecture in healthy and Salmonella-challenged broiler chickens. In the first experiment the research was targeted to investigate the effects of B. subtilis on performance, immune system, gut and lymphoid organ microarchitecture in broilers. A total of 120 d-old broiler chicks were randomly distributed into four groups, each group with three replicates containing 10 birds per replicate. The birds were fed a corn-soy-based basal diet (BD, control) or BD supplemented with 10% zinc bacitracin (ZnB), and 0.05g/kg or 0.1g/kg of B. subtilis, respectively. On d 21 and 35, six birds from each group were killed to collect blood and visceral organs (thymus, spleen, bursa of Fabricius, liver and small intestine). Parameters evaluated included growth performance, immune responses, relative organ weights, lymphoid organs and gut mucosal morphometry, intraepithelial lymphocytes (IEL) count and goblet cell histochemistry in mucosa. Results showed that the group fed 0.1g/kg of B. subtilis had superior (P<0.05) mean body weight and weight gain, and lower FCR compared to the non-supplemented or ZnB-fed groups.The BS-0.1 group revealed higher antibody titer against Newcastle disease (ND) virus and the supplemented groups against sheep RBCs (SRBCs) on d 35. Cell-mediated immune response post-phytohemagglutinin-P injection was attained (P<0.05) by birds in the BS-0.1 group at 24h, and by both the BS-0.1 and BS-0.05 groups at 48 and 72h compared to the ZnB and control groups. The BS-0.1 group gained higher (P<0.05) relative bursal weight on d 21 compared to the other groups. Compared to the control group, the liver, spleen and thymus weighed more (P<0.05) in the experimental groups on d 35. The histomorphological study revealed increased (P<0.05) thymus cortical width, and cortex/medulla ratio in the BS-0.1 group compared to the control. The area of the bursal follicles and germinal centers of the spleen also improved (P<0.05) in the BS-0.1 group compared to the control. Compared to the ZnB and control, higher (P<0.05) villus height, villus surface area and villus crypt ratio of the duodenum and jejunum were recorded on d 21, and higher (P<0.05) villus heightof the duodenum and ileum was noted on d 35 in the BS-0.1 and BS-0.05 groups. The number of goblet cells having acid mucin was significantly higher in the ileal mucosae of the BS-0.1 group chickens compared to the ZnB and control. In conclusion, B. subtilis type probiotics effectuated better growth performance, improved immune system and modulated morphology of lymphoid organs and gut mucosa in broilers. The second experiment was carried out to evaluate the effects of sodium butyrate on growth performance, immune status, organ weights and the microarchitecture of lymphoid organs and the small intestine compared to the effects brought about by an antibiotic. The cell-mediated immune response at 48 h post-phytohemagglutinin-P injection, and antibody titer against NDV and sheep RBCs on d 35 was higher (P < 0.05) in SB-1 chicks compared to those in the ZnB and control groups. Higher (P < 0.05) weight gain, and lower (P < 0.05) FCR were attained by the supplemented groups compared to the control. The thymus and spleen weighed more (P < 0.05) in the SB-1 group and bursa registered more (P < 0.05) weight in both SB groups compared to the control. On d 21, areas of the thymus medulla and the spleen germinal centers were larger (P < 0.05) in SB-1 chicks compared to ZnB and control chicks. The VH and VSA increased (P < 0.05) in the duodenum and jejunum in both SB groups on d 21, and in SB-1 on d 35 compared to the ZnB and control groups. The villus to crypt ratio was higher (P < 0.05) in the duodenum in SB-1 chicks compared to ZnB and control chicks. On d 35, VH in all segments and VSA in the duodenum and jejunum increased (P < 0.05) in SB-1 chicks compared to ZnB and control chicks. Statistically, IEL count was not significant among supplemented groups. On d 21, the number of goblet cells containing acidic mucin increased (P < 0.05) in all the segments of the small intestines in the SB-1 group compared to the control group and on d 35 in the ileum compared to the other groups. In conclusion sodium butyrate elicited better growth performance, improved immune system and modulated the morphology of lymphoid organs and the gut mucosa in broiler chickens. The third experiment was focused to assess the effect of B. subtilis and SB on gut development, growth performance and immune system in broilers challenged with S. Gallinarum. Better growth performance was reported in the supplemented groups compared to the NC-S group due to better feed efficiency. The B. subtilis-supplemented group exhibited higher (P < 0.05) cellular immunity and antibody titer against NDV compared to the PC-S and NC-S groups. Furthermore, B. subtilis¬- and SB-supplemented groups reflected higher (P < 0.05) relative thymus and bursa weights, and improved microarchitecture of the lymphoid organs compared to the NC-S group. On d 21, villus surface area in the jejunum and ileum increased (P < 0.05) in sodium butyrate-treated birds. The crypt depth of the jejunum decreased (P < 0.05) in B. subtilis and sodium butyrate groups compared to NC-S and PC-S groups. On d 35, the villus height, villus surface area and VH:CD ratio of the duodenum increased (P < 0.05) in the supplemented groups compared to the NC-S group. The FCR, Salmonella population in ceca and mortality were higher (P < 0.05) in the NC-S group. In conclusion, the prophylactic use of the B. subtilis probiotic and SB alleviated stress associated with SalmonellaGallinarum infection and improved performance, immune function, lymphoid organs and gut mucosal development in infected broilers. Further analyses are needed to reveal the mechanism(s) by which B. subtilis and sodium butyrate produce such effects. Availability: Items available for loan: UVAS Library [Call number: 2790-T] (1).

20. A Study On Point Prevalence, Etiological And Biochemical Investigations Of Post Parturient Haemoglobinuria In Buffaloes In Tehsil Bhalwal

by Muhammad Azeem (2015-VA-430) | Dr. Muti-ur-Rehman Khan | Prof. Dr. Asim Aslam | Prof. Dr. Shafqat Fatima Rehmani.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Post-parturient haemoglobinuria is a disease of great economic importance of sub-continent affecting a large number of buffaloes. It is characterized by intravascular hemolysis, haemoglobinemia, haemoglobinuria ultimately leading to anemia. The exact pathogenesis is yet unknown as there are many diversified etiological factors have been associated with this disease. All the relevant information is relatively scanty. Consequently present study has been aimed to study all possible risk factors associated with this disease in tehsil Bhalwal of district Sargodha where a large number of increasing cases were reported by the local governmental body. Etiological, hematological and biochemical risk factors were quantified to facilitate control measures and upcoming research priorities. This study was conducted from the period of about 4 months from November 2016 to February 2017. Cross-sectional epidemiological observations were documented on hemoglobinuric and healthy buffaloes for hematological and biochemical study related to parturient haemoglobinuria. The sample size was determined to three hundred and eighty four animals.Present study was observed during the period of four months (November 2016 to February 2017). Out of 384, forty animals (n=40) were confirmed with post parturient hemoglobinuria. The point prevalence observed during the period of four months was 10.4%. Buffaloes showing signs of hemoglobinuria along with parturition history, pale mucous membranes, mild tachycardia and dyspnoea was assumed as affected with post-parturient haemoglobinuria while animals suffering from other problems like babesiosis causing red urine were omitted from the study after verification of diagnosis through giemsa staining. The blood samples were processed for haematological analysis for the final confirmation of positive   haemoglobinuric buffaloes. Blood sample collected and placed in EDTA vacutainerswas processed for hematology to study hemoglobin (Hgb) values, total erythrocytes count (TEC), erythrocytes sedimentation rate (ESR) and hematocrit (Hct), total leukocyte count (TLC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) in addition to mean corpuscular hemoglobin concentration (MCHC) by using haematological analyzer. Haematological analysis of all the samples was made from Department of Pathology, UVAS, Lahore.Serum samples of all buffaloes were analyzed for biochemical analysis asalkaline phosphatase (ALP), serum urea, glucose,bilirubin, creatinine, calcium, phosphorus, copper, and molybdenum. Moreover, urinalysis was done for gross and biochemical analysis. Results of the study revealed significant difference among complete blood count (CBC) includingHgb, TEC, Hct and TLC, ESR, MCV and MCH. However, there was no significant variation among MCHC values in affected buffaloes. Serum biochemistry also revealed significant difference of various parameters including ALP, creatinine,BUN, total bilirubin, phosphorus, copper and molybdenum. However, no significant difference was detected among the healthy and affected groups regarding blood glucose and serum calcium levels. There was significant elevation in pulse and respiration rates in buffaloes affected with hemoglobinuria. The results regarding mineral analysis of the soil shows significant difference in phosphorus and copper. Moreover, mineral levels of soil and serum of animals showed significant relation of phosphorus levels, followed by the levels of molybdenum. Calcium and copper levels also showed moderate relationship. Observations regarding parity/lactation number reveal the highest incidence rate of 35% among buffaloes at 3rd lactation, followed by buffaloes at 4th, 2nd, 5th, 1st and 6th lactation, respectively. Milk production showed direct relationship with buffaloes affected with post parturient hemoglobinuria. From the present study, it is concluded thathemoglobinuria was observed in buffaloes of tehsil Bhalwal may be due to variation of soil composition particularly the deficiency of Phosphorus which may lead to the lysis of erythrocytes and hemoglobinuria through various pathways. However, efficient replenishment of minerals content in fodder producing soil is necessary to overcome the disease in buffaloes affecting from parturient hemoglobinuria in the aforementioned area.   Availability: Items available for loan: UVAS Library [Call number: 2847-T] (1).

21. Pathobiological Investigations Of Peste Des Petits Ruminants (Ppr) Virus With Reference To Antiviral Activity Of Nigella Sativa (Black Seed)

by Kiran Aqil (2008-VA-456) | Dr. Muti Ur Rehman Khan | Prof. Dr. Asim Aslam | Dr. Aqeel Javeed.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: Peste des Petits Ruminants (PPR) is a highly contagious, infectious, acute or sub-acute transboundary viral disease of domestic and wild small ruminants. It is an economically important viral disease of sheep and goats causing varying degree of morbidity and mortality in susceptible animals which may be as high as 100 and 90 per cent, respectively. PPR is responsible for serious socioeconomic problems. There is no data available regarding pathogenesis and field virus characterization to compare it with vaccinal strain for any difference. Nigella sativa(Black Seed) has antiviral activity against many viruses. Therefore present studywas undertaken to investigate the antiviral effect of Black Seed in vivo and in vitro against PPR virus. Further more time course detection of virus is still needed to be studied.  Nigella sativa (Black seed) has antiviral activity against PPR virus.  Pathogenesis can better be studied through histopathology, necropcy findings and morphometric changes. A total of 250 clinically positive samples suspected for PPR virus were included in the study. Samples were consisted of nasal, ocular and anal swabs; whole blood in EDTA were collected from suspected animals. In case of mortality morbid material included lungs, liver, spleen and mysenteric lymph nodes were included in the study. Samples were subjected to immune capture Elisa for detection of viral antigen in suspected samples. Samples which found positive foe IC – Elisa were then subjected to RT-PCR for confirmation of virus. After confirmation of virus through IC – Elisa and RT-PCR the positive samples were subjected to virus isolation on vero cell. After isolation of virus, the TCID 50 of the virus was calculated for preparation of inoculum for further use. In this experiment mesenteric lymph nodes and spleen found to be major organ for isolation of PPRV.RT-PCR found to be most reliable and confirmatory diagnostic test for PPRV. Field Virus adaptation on vero cells found to be difficult to optimize. In this experiment antiviral activity of black seed was checked on vero cells infected with PPRV. Three extracts of N. Sativa were prepared to check the in vitro antiviral activity of black seed. In this study poly saccharides extracted from black seed found to be more effective against PPRV. Adaptation of field virus was done on Vero cell line. Antiviral activity of Black Seed extract was determined in vitro on Vero cell on bases of CPE (Cytopathic effect). The ethanolic and aqueous extract were found to be more toxic to consistency of monolayer of vero cells. The TCID50 of virus was calculated after treating cells with different extracts. In this study poly saccharides extract exhibit lower TCID50‘s as compared to ethanolic and aqueous extract which showed higher TCID50’s.So less cytopethic effect was observed in vero cells treated with black seed extracts. Antiviral activity was determined on base of CPE. Pathogenesis of virus in natural host was studied through time course detection of virus in body secretions, blood, organs. Histopathological changes were studied.20 goats were procured from market divided into four groups (n=5) A,B,C and D. In animals of group A prophylactic effect of N.Sativa was studied. In group B complete pathogenesis of PPR virus was studied without any prophylactic or therapeutic measure. In group C therapeutic effect of N. Sativa was studied after onset of clinical picture of disease. At the end of this experiment, clinical picture, gross pathology, histopathology, and morphometric changes revealed that N. Sativa has noticeable prophylactic effect on PPR infected goats. It can be used as a therapeutic agent in PPR infected goats but it can’t control pathological effect of virus after onset of infection. SUMMARY 130 Data collected were statistically analyzed by using Microsoft Excel (Microsoft Excel, 2007) and SPSS (for Windows, Version 16.0). The data were put the descriptive analysis and Chi square test was employed to test the significance and test of hypotheses It was concluded that Black Seed therapy possessed marvelous prophylective effect against PPR virus and RT-PCR was the most efficient methodology to confirm the virus. Availability: Items available for loan: UVAS Library [Call number: 2890-T] (1).

22. Pathological Association Of Nramp 1 Genotypes With Brucella Resistance And Susceptibility In Diseased And Non Diseased Cattle

by Muhammad Zaheer Iqbal (2005-VA-61) | Dr. Raheela Akhtar | Prof. Dr. Asim Aslam | Prof. Dr. Kamran Ashraf.

Material type: book Book; Literary form: not fiction Publisher: 2017Dissertation note: A total of 200 cattle were divided into five groups including: Group A Sahiwal cattle, Group B Jersy cattle, Group C Frisian cattle, group D Sahiwal cross Jersy and group E Sahiwal cross Frisian. Out of total of 200 serum samples from suspected cattle we found 155 samples positive by RBPT and 109 were positive for Brucella abortus by PCR. Comparison of presence of Brucella abortus was statically made in all five groups using chi square. The study was conducted on 200 animals of five breeds including Sahiwal, Jersey Cross Sahiwal, Frisian Cross Sahiwal, Fresian and Jersey around farms of Punjab. Blood sample (3mL) was collected in EDTA vaccutainers from each animal. Serum from blood samples were collected in 3 mL eppendorf tubes with the help of sterile pipettes. Serum samples were screened out for brucellosis by Rose Bengal Plate Test (RBPT). RBPT positive samples were stored at 40C for further processing. Rose Bengal Antigen was purchased from Veterinary Research Institute (VRI), Lahore. The antigen was stored at 40C in refrigeration during the study according to manufactures recommendation. DNA quantification was performed on “Thermo Scientific NanoDrop spectrophotometer ND-2000” and by running extracted DNA on 0.8% agarose gel. PCR for amplification was done with a total volume of 20 μL by adding primer pair and extracted DNA to the PCR master mix in following concentrations. 2ml of forward and reverse primer was taken respectively. 4ul of PCR grade water was added and DNA was taken in 2 ul quantity. The total volume of master mix obtained was 10 ul. The thermocycler (Bio-Rad, CFX ™, Singapore) was set with the conditions for the total of 35 cycles of PCR. For optimization process of primers different options for PCR reaction mixture Summary 41 and PCR cyclic conditions were tried for two objectives. To get maximum amplification, by using minimum volume of chemicals. Changing the volume of magnesium chloride, deoxynucleotide triphosphate (d NTPs) and Taq polymerase, amplification can be increased. Primers annealing temperature is considered critical for optimization. Denaturation of DNA samples were performed at 94 0C for 5minutes. Annealing step was performed at 63 0C for 30 second. Then, initial extension of DNA samples was conducted at 72 0C for 30 second. Finally, extension was performed at 72 0C for 5 minutes to get results. The PCR product was confirmed by running the PCR product on 0.8 % agarose gel electrophoresis was performed at 100 Volts for 30 minutes. The NRAMP1 gene encodes a divalent cation transporter, located in the phagolysosomal membrane of macrophages, which has been associated with resistance to intracellular pathogens. In cattle, natural resistance against brucellosis has been associated with polymorphisms at the 3′ untranslated region (3′UTR) of the NRAMP1 gene, which are detectable by single-strand conformational analysis (SSCA). Genetic selection of domestic animals resistant to pathogens has been applied mostly to farm animals, particularly cattle. Identification of genes linked to natural resistance may allow for a better understanding of natural resistance with obvious practical implications. These genes may also function as markers for prediction of genetic resistance against specific diseases. Recommendations: From this study we concluded that Nramp1BB gene is resistant to brucellosis, while Nramp1 AA is susceptible to brucellosis. Summary 42  By gene knock out technique breeds resistant to brucellosis can be produced.  Criss Crasper technique can be used for gene knock out process. Availability: Items available for loan: UVAS Library [Call number: 2953-T] (1).

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