51.
Immuno-Modulatory Effects Of Beta-Glucans And Nucleotides Based Commercial Products In Broiler Chicken Experimentally Infected With Velogenic Newcastle Disease Virus
by Hafiza Zain ul Fatima (2008-VA-232) | Prof. Dr. Asim Aslam | Dr. Raheela Akhter | Dr. Aamir Ghafoor.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Immune-potentiation effect of commercially available products containing beta-glucans (betaplex and catalyzer) and nucleotides (electroimmune and avimmune) in the broilers was evaluated. A total of 120 broiler chicks were reared under standard husbandry conditions. The birds were vaccinated by live NDV vaccine at 5, 14 days and killed Newcastle disease virus (NDV) vaccine at 7 days of age. Additionally, live infectious bronchitis and infectious bursal disease vaccines were also given through water at 5 and 10 days of age, respectively. The birds were randomly divided into six groups each comprising 20 birds. Four groups (treatment groups) were offered four commercial immune-booster products at 7, 14 and 21 days of age for 72 hours. Fifth group (untreated vaccinated) was not offered any drug while sixth group (untreated and unvaccinated) was not given any drug and also remained unvaccinated against NDV. Anti NDV antibodies were measured at 7, 14, 21 and 28 days of age and heterophil/lymphocyte (H/L) ratio was measured at 14, 21 and 28 days. Weight of immune organs and performance of birds was also measured from each group. Finally, 15 birds from each group were inoculated with virulent NDV through intranasal route at 28 days of age to assess the protection against challenge infection.
Mean GMT anti NDV antibody titers in vaccinated groups were 3.65 (±0.59) at one week of age which were raised to the levels of 5.85 (±0.88), 5.50 (±1.00), 6.10 (±0.64), 5.90 (±0.91) and 5.45 (±1.05) in the betaplex, catalyzer, electroimmune, avimmune, and untreated control groups, respectively. Maximum antibody titer was achieved in electroimmune treated group while minimum in the catalyser treated ones. In non-vaccinated control group the antibody titers were minimum in the beginning that reduced to negligible level at four weeks of age. The H:L ratio of
Summary
48
all treatment groups did not significantly vary at 14 days of age while at 21 and 28 days of age, the birds receiving electroimmune have significantly low H:L ratio in comparison to other treatment groups. Weight of liver and spleen in nucleotide treated groups was significantly higher than other treatment and control groups while there was no significant difference in the weight of bursa of fabricious in all treatment groups. Relative weight of all immune organs to the live body weight did not significantly vary in all treatment and control groups. Performance of the birds as a measure of feed conversion ratio (FCR) was highest in electroimmune treated birds. The challenge protection data shows that minimum mortality of 40 % (6/15) was observed in nucleotides treated groups while betaplex and catalyzer treatment groups have 47 % (7/15) and 53 % (8/15) mortality ratios in comparison to 60 % (9/15) mortality in untreated birds. Results of present study suggest that nucleotide or beta glucans supplementation in the diet have beneficial effects to enhance the humoral immune response and better bird performance in broilers. Use of these products not on delay the onset of clinical disease but also helpful in providing better protection against challenge to velogenic NDV. Availability: Items available for loan: UVAS Library [Call number: 2516-T] (1).
52.
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: ; 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).
53.
Amelioration Of Pathological Changes Due To Infectious Bursal Disease By The Administration Of Mentofin And Asi-Mirus In Broiler Chicken
by Muhammad Umair Shah (2011-VA-15) | Prof. Dr. Asim Aslam | Dr. Ghulam Mustafa | Dr. Ali Ahmed Sheikh.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Poultry industry is the second largest industry in Pakistan but despite of its rapid growth rate it is facing huge economic losses due to many infectious diseases. Infectious Bursal Disease is one of them. Huge economic losses in case of infectious bursal disease are due to immunosuppression and high mortality.
In Pakistan, commercially available vaccines are abruptly used to control different viral diseases but unfortunately failure of these products occur from time to time. Hence, current study was designed to determine the immunostimulatory effect of two commercially available products (Mentofin and ASI-MIRUS) against IBD vaccine.
A total 300 broiler chicks were taken, divided into six groups each having 50 birds and were replicated under controlled conditions. A, B and D groups were vaccinated with the IBD live virus vaccine. B and C groups were treated with Mentofin. D and E groups were treated with ASI-MIRUS while F group served negative control. To detect antibody titer against IBDV at every week (0-42 days of age), a commercial ELISA kit, IDEXX Flock Chek standard (IDEXX Corporation, Westbrook, ME, USA) was used. In order to analyze gross and microscopic changes in bursa, postmortem examination and histopathology of bursa was done.
The volatile oils in Mentofin and ASI-MIRUS have effective immunomodulatory effects on humoral immune response in broiler chicks. Eucalyptus and peppermint oils increase bursa to body weight (B/BW) ratio as compared to untreated birds. Results of present study indicated the highest antibody titer in group D supplemented with ASI-MIRUS and vaccinated as compared to group B supplemented with Mentofin and vaccinated. Significantly high bursa to body weight ratio also observed in vaccinated group D (ASI-MIRUS treated) comparing with other
CHAPTER 6
SUMMARY
Summary
35
vaccinated groups A and B. In Group B (Mentofin treated), bursal samples showed necrosis at medullary region of bursal follicle. Group D (ASI-MIRUS treated) showed the active follicle consist of lymphoid cells and shown no obvious histopathological lesion. So present study showed that ASI-MIRUS is reduced the severity of IBDV which has more beneficial effect on immune response against IBD vaccinated Broiler Chicken as compared to Mentofin. Availability: Items available for loan: UVAS Library [Call number: 2557-T] (1).
54.
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: ; 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).
55.
Pathological Investigation And Molecular Detection Of Avian Pathogenic E.Coli Serogroups In Broiler Birds
by Muhammad Azeem Riaz (2008-VA-132) | Prof. Dr. Asim Aslam | Dr. Muti Ur Rehman Khan | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: The present study was designed to identify the serogroups present in field and to study their pathological effects in experimentally infected broiler chicks. The present study was attempted to scan the rfb gene clusters in APEC predominant serotypes O1, O2 and O78 strains and to develop Multiplex PCR method for serotyping of the O-antigens. The Multiplex PCR method was used for the identification of serotypes of APEC. The second part of the study was to study the pathological lesions caused by most prevalent serogroup in experimentally infected broiler chicks.
A total of 100 tissue samples (lungs and livers) were collected from colibacillosis suspected broiler birds. Streaking was done from these samples on three different media and it was found that 80% isolates were positive on MacConkey media, 60% were positive on EMB media and 40% were found pathogenic for E.coli on Congo red media.
The colonies which were of pink color on congo red media were considered as pathogenic. DNA was extracted from these colonies by boiling method by picking single colony from each petri plate. Extracted DNA was further used for PCR to confirm the three serogroups i.e O1, O2 and O78.
The PCR results showed that 8% isolated samples were found as pathogenic as O2 strain was found dominant among all. Only two genomic DNA samples were found of O1 serogroup After confirmation of serogroups inoculum of Avian pathogenic E.coli O2 strain was prepared to experimentally infect the broiler birds. Birds were infected at the age of day 7 via intratracheal route.
Following the experimental infection of birds, they were monitored for any pathological lesions which were not present significantly while some birds were off feed, reluctant to move, head down posture and were keeping themselves in isolation.
Summary
46
Postmortem of dead birds was performed and pathological lesions were noted. Livers were found to be congested, enlarged and white fibrinous layer over liver was present. Lungs were also affected with the disease and white layer was present on lungs too. Lungs were consolidated and congested.
Histopathology of lungs and livers was performed. It was noted that there was mononuclear cells infiltration and thin fibrinous layer over liver. Thickening of the liver capsule was noted due to infiltration of mononuclear cells and there was marked congestion in hepatic portal areas and the central vein. There was atrophy of adjoining hepatic cords due to greatly distended and congested sinusoids. Besides these changes, hepatocytes in various stages of degeneration along with hemorrhages, areas of congestion and fatty changes in a few places could be seen.
There was infiltration of heterophils, severe congestion, lymphocytes and macrophages in the wall of the bronchus as well as in the peribronchial alveoli. There was marked presence of granuloma in lungs. Some birds displayed thickening of the pleura and consolidated areas covered with yellowish fibrin in lungs.
The experimental infection of avian pathogenic E.coli confirmed the hypothesis that it causes pronounced histopathological lesions in broiler birds. Availability: Items available for loan: UVAS Library [Call number: 2591-T] (1).
56.
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: ; 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).
57.
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: ; 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).
58.
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: ; 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).
59.
Comparison Of Conventional And Modern Diagnostic Techniques For Bovine Tuberculosis With Associated Risk Factors
by Arslan Tariq (2014-VA-960) | Prof. Dr. Asim Aslam | Dr. Yasin Tipu | Prof. Dr. Mansur-Ud-Din Ahmad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2016Dissertation note: Bovine tuberculosis is a chronic bacterial disease of cattle that occasionally affects other species of mammals. It is a zoonotic disease that can be spread to humans, typically by the inhalation of aerosols or the ingestion of unpasteurized milk. The present study was conducted to diagnose tuberculosis in cattle and buffalo by using conventional method of Tuberculin. Later on, the positive samples of tuberculin test were tested on ELISA and PCR. Additionally, associated risk factors including the age, sex, breed and body condition were also investigated.
One hundred and ninety-two sexually mature animals of age 2-8 years for detection of tuberculosis were selected randomly from four organized farms in eastern wing Lahore district. The animals were divided into two main experimental groups labeled as group B (Buffalos) and group C (Cows). Each group was further sub-divided into 3 sub-groups namely B1, B2, B3 and C1, C2, C3 with age group of below 4y, 4-6y and over 6y respectively.
On the basis of tuberculin test number of positive cases of bovine tuberculosis in buffalo were 7.29 % while in cattle were 11.46% out of total 96 animals of each group. On the basis of diagnosis with ELISA-IFN-γ assay, percentage of positive cases were 71.43% and 72.73 % respectively out of tuberculin positive samples. Whereas the percentage of positive cases in case of PCR of buffalo and cattle were 85.71% and 90.91% respectively.
Number of positive cases in buffalo at age below 4 years, 4-6 years more than 6 years were observed as 1, 2 and 4 respectively. While in cattle, number of positive cases were found to be 2, 3 and 6. In case of buffalo, number of positive cases of bovine tuberculosis in male and female were 2 and 5 respectively. In cattle, 4 males and 7 females were declared as positive. In case of buffalo, number of positive cases of bovine tuberculosis in Nili Ravi (pure breed) and non-descript breeds were 2 and 5 respectively. Number of positive cases in case of indigenous and exotic breeds
Summary
41
of cattle were 3 and 8 respectively. In buffalo, only 1 animal with good body condition was found positive against bovine tuberculosis, while 2 animals with fair body and 4 animals with poor body condition were observed as positive cases. In case of cattle, only 1 animal with good body condition, 3 with fair body condition while 7 with poor body condition were declared as positive cases.
It can be concluded from the study that Modern techniques are more specific than conventional techniques. It is also deduced from study that cattle are more susceptible to bovine tuberculosis rather than buffalo. Moreover, female animals were more prone to infection than that of male, older animals were found to be infected more frequently than that of younger animals. Animals with poor body conditions were at high risk of infection than animals with good body conditions. Availability: Items available for loan: UVAS Library [Call number: 2648-T] (1).
60.
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: ; 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).
61.
A Comparative Study Of Non-Antibiotic Feed Additives On Experimental Colonization Of Salmonella Enterica Serovar Enteridis And Intestinal Pathomorphology In Broiler Chickens
by Adeem Rehman Raffie (2010-VA-233) | Prof. Dr. Asim Aslam | Dr. Muhammad Yasin Tipu | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: The utility of antimicrobial agents as a preventive measure has been questioned, given extensive documentation of the evolution of antimicrobial resistance among pathogenic bacteria. Non-antibiotic feed additives (probiotics, prebiotics, essential oils and organic acids) are being considered to fill this gap and already a few farmers in the country are using them with good results. The present study enable us to understand and compare the beneficial effects of non-antibiotic feed additives on salmonella enterica colonization and changes in intestinal morphology.
This study is designed to evaluate the effect of non-antibiotic feed additives on salmonella enterica serovar enteritidis colonization in intestine of broilers chickens and compare the intestinal morphology between normal healthy, non-antibiotic feed additives supplemented and salmonella challenged broiler chickens.
A total of 125 commercial day-old broiler chicks were procured from the local market. The chicks were divided into six groups A (Basal diet, negative control group), B (Challenge + Basal diet, positive control group), C (probiotic + Challenge + Basal diet), D (prebiotic + Challenge + Basal diet), E (essential oils + Challenge + Basal diet) and F (organic acids + Challenge + Basal diet) with 20 chicks in each group and given separate treatments.
Two separate experiments were carried out for salmonella recovery from cecal tonsils and intestinal pathomorphic evaluation. Villus length, villus width, villus surface area and crypt depth were measured by micrometery. The collected data from both experiments was analyzed using the statistical technique of comparing more than two groups i.e. Analysis of variance (ANOVA) through SPSS 16.0.
Summary
45
There was an overall increase in all the parameters of intestinal morphometric analysis for all the treatment groups except for the control negative group which showed lowest values. Maximum villus height of 1794.2±63.96 μm in duodenum was achieved by group E, which was given essential oils. Whereas maximum villus surface area index of 1662.6±389.16 mm2 was recorded in group D, which was treated with prebiotics. Maximum villus height of 940.35±23.96 μm and surface area index of 568.92±36.27 mm2 in ileum mucosa was recorded in group D, treated with prebiotic. . Recoverable salmonella was most reduced by probiotics and organic acids.
Final results show that there is an overall increase in histological parameters of the mucosa of duodenum and ileum in the groups fed non-antibiotics feed additives as compared with control groups. Prebiotics showed the maximum positive effects in histological parameters whereas probiotics showed maximum positive effect for decreased recoverable salmonella count. Hence this study suggests that a combination of non-antibiotic feed additives will be beneficial for the intestinal health of broiler chickens but there is a need for more research on combinations of non-antibiotic feed additives. Availability: Items available for loan: UVAS Library [Call number: 2844-T] (1).
62.
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: ; 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).
63.
Detection Of Genetic Variants In Interferon Gamma Gene And Its Association With Resistance Against Mycobacterium Bovis In Buffalo
by Awais Nawaz (2010-VA-219) | Prof. Dr. Asim Aslam | Dr. Muhammad Yasin Tipu | Dr. Jawad Nazir.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Bovine Tuberculosis (bovine TB) is a chronic disease of animals and has been known for the significant zoonotic impact. Immune mechanisms necessary for protection against Bovine TB are poorly understood. Interferon-γ cytokine has been reported critically and it is important to study its role in immunity against Bovine TB.
Blood samples were collected from 100 Animals from Peri-urban areas of Lahore, Gujranwala and Okara, Pakistan. Genomic DNA was extracted from the samples. Specific primers were designed to amplify specific portion of IFN-γ gene. Amplified products were sequenced and analyzed by bioinformatics tools. Interferon-γ assay was performed from blood collected in heparin coated vacutainers for the quantification of interferon-γ cytokine in different groups of animals. Blood samples from mycobacterium infected symptomatic and symptomatic animals were processed in Haematology analyzer for complete blood count.
Genetic sequencing of bovine Interferon gamma gene (IFN- γ) help in finding out the Genetic Variations to characterize its role in resistance against Mycobacterium bovis infection. This study help in finding out the confirmed markers for natural resistance against bovine TB that can be used in future selection and breeding programmes. The comparison of hematological values and Interferon gamma level of different groups of animals help us for the detailed diagnosis and prognosis of the disease.
The collected data from hematological analysis of Mycobacterium infected symptomatic animals (Group A), Mycobacterium infected asymptomatic animals (Group B) and non-infected animals/control Group (Group C) was analyzed using the statistical technique of comparing more than two groups i.e. Analysis of variance (ANOVA), One way ANOVA through SPSS 16.0.
CHAPTER 6
SUMMARY
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Mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration were found non-significant (p>0.05). White blood cells, Lymphocytes, Platelets, Mean platelet volume and Mean corpuscular volume were found significant (p<0.05). Granulocytes, Red blood cells and Red cell distribution width values were found highly significant.
Interferon gamma assay provided confirmation about the presence of disease in the animals by indicating interferon gamma level to insight the undergoing pathogenesis which was helpful in the detailed diagnosis of the disease. Later on it helped us in the confirmation of false positive results by Tuberculin test.
Final results revealed four intronic variations in different groups of animals. Three of them were found in Group A and B and one was found in Group C (non-infected animals) by Primer 1 (P1). Intronic variations don’t have significant effect but they may have an impact on the regulation of the gene.
We found Transversions (T > A), (A > T), (T > G) were found in mycobacterium infected symptomatic group of animals (Table: 4.7). Transversion (C > G) at and deletion (G >_) was found in this group and exclusive presence of these SNP’s in this group can be considered significant and responsible for the infection. Transversion (A > C) and addition (_ > G) were found in mycobacterium infected asymptomatic group of animals. These two SNP’s are significant as they have been found only in this group. We can infer that the presence of these two SNP's is responsible for the infection along with making them asymptomatic towards the disease.
It was noted that Transition (G > A), (T > C) has been found common in mycobacterium infected symptomatic and asymptomatic group of animals. This common mutation at same position in both groups is quite significant and could be attributed to the occurrence of disease.
Summary Availability: Items available for loan: UVAS Library [Call number: 2881-T] (1).
64.
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: ; 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.
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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).
65.
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: ; 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
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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
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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).
