1.
Comparative Studies On The Sensitivity Of Polymerase Chain Reaction (Pcr) And Conventional Serological Methods For the Diagnosis of Bovine Brucellqsis
by Raheela Akhtar | Prof. Dr. Zafar Iqbal Chaudhary | Prof. Dr. Abdul Rauf Shakoori | Prof. Dr. Asim | Faculty of Veterinary Sciences.
Material type: ; Format:
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; Literary form:
not fiction
Publisher: 2007Dissertation note: The polymerase chain reaction was standardized and its efficacy was evaluated against Rose Bengal Plate Test (RBPT) and Milk Ring test (MRT) for the diagnosis of brucellosis in 200 cows and buffaloes from Lahore and Okara districts of Punjab. Under aseptic measures 200 serum and 200 milk samples were tested by RBPT, MRT and PCR on both milk and serum samples in both cows and buffaloes as described in materials and methods. RBPT showed high sensitivity values (27.7% in cows and 45.2% in buffaloes) than serum PCR (25% in cows and 3 9.6% in buffaloes) but on other hands MRT showed low sensitivity (11.1% in cows, 25.4% in buffaloes) and high specificity (98.4% in cows and 93.6% in buffaloes) than milk PCR with sensitivity of 13.8% in cows, 29.4% in buffaloes and specificities of 95.2% in cows and 89.3% in buffaloes respectively. The comparison of PCR assays conducted on both types of samples showed high sensitivity of serum PCR against milk PCR. The comparison of RBPT and MRT in both species showed high sensitivity of RBPT than MRT. But due to low positive predictive value of RBPT and instability in its results in both species it is concluded that there is no significant difference in PCR and serological methods so no single test can be used for the exact diagnosis of bovine brucellosis.
Availability: Items available for loan: UVAS Library [Call number: 0957,T] (1).
2.
Tissue Tropism Of Velogenic-Viscerotropic Newcastle Disease Virus In Broiler Chickens
by Tasra Bibi | Prof. Dr. Asim Aslam | Dr. Raheela Akhtar | Prof. Dr. Tahir.
Material type: ; Format:
print
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drama
Publisher: 2014Dissertation note: ND is an infectious, highly contagious and widespread disease of avian species and has a considerable economic impact on poultry industry. This study is a preliminary work to understand the mode of action of the recent VVNDV isolate of the UVAS, towards the tissue tropisms of both lymphatic and non lymphatic organs. One hundred chicks purchased from the local hatchery and reared for the trial including control, however, the VVNDV strain was received from the QOL, UVAS, Lahore, Pakistan. Three trials were conducted using the challenge dose 100,000 ELD50 (Group C) and 10,000 ELD 50 as (Group B) and 1000 ELD 50 as (Group A). Five chickens were selected randomly from each group and slaughtered on daily basis, including two chicks from control. These samples were used for histopathology and immunohistochemistry (IHC) test. Conclusively, the study indicates that NDV induces early necrosis in the lymphoid tissues of infected chickens which is correlated with the severity of the disease caused by each dilution. Necrosis does not seem only to be the direct viral replication and indirect effects may lead to death of the animals, due to depletion of lymphoid organs. However, the peak hours were recorded 72 hours to 96 hours post infection in all lymphoid and non lymphoid organs irrespective of the dilution factor of the VVNDV. Immunohistochemistry is not a routine practice in diagnostic test, however, this study may lead to a roadmap in understanding the interpretation of the clinical/pathological picture and the tropism may be helpful in future to study some other aspects like failure of commercial vaccines and to control the outbreaks of NDV in the country an endemic as well as a devastating disease of the poultry industry.
Availability: Items available for loan: UVAS Library [Call number: 1824,T] (1).
3.
Study Of Carrier Potential Of Newcastledisease Virus In Common Wild Bird Species
by Tahir Munir | Prof. Dr. Asim Aslam | Dr. Muhammad | Dr.Ishtiaq Ahmed.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 1892,T] (1).
4.
Optimization Of Multiplex Pcr For The Simultaneous Detection Of Babesia Caballi Theileria Equi And Trypanosoma
by Muhammad Zeeshan Khalid | Prof. Dr. Asim aslam | Dr. Yasin tipu | Prof. Dr.Habib ur rehman.
Material type: ; Format:
print
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2006,T] (1).
5.
Optimization Of Loop Mediated Isothermal Amplification Lamp Technique For The Molecular Diagnosis Of Surra In Domestic Animals
by Muhammad Saleem iqbal | Dr. Haroon akbar | Dr. Muhammad lateef | Prof. Dr. Asim.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2039,T] (1).
6.
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: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2070,T] (1).
7.
Comparisons Of Histopathological Effects Of Live Vector Immune Complax And Intermediate Plus Vaccines Of Infectiors Bursal Disease on the Bursa of Fabricius of Commercial Broiler
by Rana Waqas Arshad | Prof. Dr. Asim Aslam | Dr, Muhammad Saeed Imran.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2086,T] (1).
8.
Study Of Carrier Potential Of Salmonella Infections In Common Wild Birds Species
by Mutahir Ali Mir | Prof. Dr. Asim Aslam | Dr. Ishtiaq Ahmad | Dr. Muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2148,T] (1).
9.
Molecular Characterization Of Brucella Abortus Strains In Bovines
by Muhammad Ramzan | Dr. Raheela Akhtar | Prof. Dr. Aneela | Prof. Dr. Asim Aslam.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Abstract Availability: Items available for loan: UVAS Library [Call number: 2169,T] (1).
10.
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: ; 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
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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).
11.
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: ; 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).
12.
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: ; Literary form:
not fiction
Publisher: 2015Dissertation note: CD Error Availability: Items available for loan: UVAS Library [Call number: 2264-T] (1).
13.
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: ; 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).
14.
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: ; 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).
15.
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: ; 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).
16.
Study On Pathogenesis Of Mixed Infection Of Infectious Bronchitis Virus (Ibv) And Avian Influenza Virus (H9n2) In Experimentally Infected Broiler Chicken
by Arsala Khan (2013-VA-605) | Prof. Dr. Asim Aslam | Dr. Muhammad Yasin Tipu | Prof. Dr. Khushi Muhammad.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2015Dissertation note: Avian influenza and infectious bronchitis are the most devastating contagious and viral diseases, affected poultry in Pakistan and many other Asian countries. Regarding the virulence, Avian influenza may be low, or high pathogenic. Mainly due to its pandemic potential and contagious nature, the low pathogenic avian influenza virus (H9N2) is of major concern in poultry industry especially in Pakistan. Co-infection of poultry with more than one bacterial and /or viral agent is common in field especially in our country due to inadequate biosecurity measures which causes high mortality and confusing the diagnosis of these viruses. Hence the present project was designed to study the pathogenesis of mix infection of IBV and H9N2 through gross, histopathological lesions and antibody titer in experimentally infected broiler chicken. For this purpose a total of 80 specific pathogen free chicks were procured, randomly divided into four groups A, B, C and D; and raised under uniform managmental condition in experimental sheds of UVAS Lahore. Group A and B were intarnasally challenged with 0.2 ml infectious bronchitis virus (EID50 = 104.5 per 0.1 ml) at 23rd day of age, likewise group B and C were inoculated with 0.2 ml of H9N2 (EID50 = 106 per ml) at 26th day of age while, group D served as a negative control group. After infection the clinical signs, feed consumed and body weight gain were regularly monitored, the serum samples taken on days 23rd, 26th, 29th, 35th, and 40th day of age were tested for HI antibody titer simultaneously, the necropsy of birds (n=3) sacrificed were conducted to observe the gross lesions, tissue samples including lungs, liver, kidneys and trachea were collected for histopathological evaluation. In IBV infected group, respiratory distress i.e., tracheal rales, nasal discharge, coughing, sneezing, gasping and reduced feed intake were observed during early stages, later up to 10 days post infection watery diarrhea with ruffled feathers were observed. In mix infected group clinical signs manifested rapidly and were persistent with
Summary
59
high severity. Gross lesions in mixed infection were more profound, including; severely congested and hyperemic lungs, tracheitis with catarrhal exudates in lumen; pale, swollen and enlarged kidneys with urates deposition in tubules. Six birds died in mix infected group, revealed caseous exudate in trachea extended up to lower bronchi while, in IBV infected group lesions were mild and confined to trachea, lungs and kidneys. Mortality was high in mix infected group (30%) followed by IBV infected group in which two birds died. While, in H9N2 infected and control group none of the bird died. Histopathological lesions in mix infected group were aggravated markedly tracheal epithelium degeneration and sloughing; congestion, interstitial nephritis, leukocytes infiltration, tubular degeneration and necrosis were observed while, in lungs pneumonia of peribronchiolar area and interstitium with lymphocyte and macrophages infiltration, additionally degeneration and vacuolization of hepatocytes with focal necrotic areas in liver were also noted. In IBV and H9N2 infected group microscopic lesions were of mild degree. GMTs against both IBV and H9N2 in mix infected group were significantly different at P>0.05 than individually infected birds. Among the groups, statistically significant increase in FCR of birds in mix infected group was observed. On the basis of these findings it might be conclude that mix infection of IBV and H9N2 causes severe disease as compared to single infection. Availability: Items available for loan: UVAS Library [Call number: 2296-T] (1).
17.
Study On The Pathogenesis Of Co-Infection Of Infectious Bronchitis (Ibv) And Escherichia Coli (E. Coli) In Experimental Chickens
by Sohail Khan (2013-VA-606) | Prof. Dr. Asim Aslam | Dr. Muti Ur Rehman Khan | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Infectious bronchitis and colibacillosis are infectious diseases affecting chicken of all ages and breeds. They are of major economic importance in commercial chicken flocks, causing huge losses. As both in humans and animals it is well documented that preceding respiratory infection of virus predispose individual to bacterial infection. Moreover, mix infection in poultry which occur when different organisms simultaneously invade birds is a major threat to poultry industry causes highly epidemic debilitating disease with high mortality which eventually leads to economic catastrophe. In recent past prevalence studies of field, E. coli had been reported with high prevalence and exaggerated disease along with other respiratory pathogens, additionally IBV had also isolated from same flocks in same season. Although a plethora of pioneering work had been done on IBV and E. coli in the previous decades but still a window in time exist in revealing there co-infection. Looking to field scenario in our country, the present study was designed to study an ideal challenge model for IBV and E. coli, by reproducing the natural infection.
80 SPF day old broiler birds were arranged into four groups, (A, B, C and D). Each group was comprised of 20 birds. Group D served as uninoculated control while, Group A and B were challenged with IBV on 23rd day of trails, and Group B and C were inoculated with E. coli infection on day 26th. Birds, (n=3) from each group were slaughtered on various days post infection, gross and histopathological lesions were observed and serum samples for HI were taken, throughout experiment. Variable clinical signs were recorded in various groups. In IBV infected group, respiratory distress i.e., tracheal rales, coughing, sneezing and gasping were noted during early stages, later up to 10 days post infection watery diarrhea with ruffled feathers were observed. In mix infected group clinical signs manifested rapidly and were persistent with high severity. Gross lesions in mixed infection were more profound,
Summary
56
including; airsacullitis, tracheitis with catarrhal exudation throughout respiratory tract; severe sepicemic lesions i.e. perihepaitis, pericarditis, pneumonia and polyserositis with swollen and pale kidneys distended by urates. 5 birds died in mix infected group revealed ascites with asphyxiation of trachea with caseous exudate. While in IBV infected group lesions were mild and confined to trachea, airsac and kidneys. Mortality was high in mix infected followed by IBV in which two birds died. While in E. coli and control group mortality were not noted. Histopathological lesions in mix infected group were aggravated markedly tracheal epithelium degeneration, deciliation and sloughing; congestion, interstitial nephritis, leukocytes infiltration, tubular degeneration and necrosis while were observed. In lungs, pneumonia of peribronchiolar area and interstitium with lymphocyte and macrophages infiltration, additionally degeneration and vacuolization of hepatocytes with focal necrotic areas were also noted. In IBV and E. coli group microscopic lesions were of mild degree. GMT of both IBV and mix infected birds were high but were not significant different (P>0.05). Among the groups, statistically significant increase in FCR of birds in mix infected group was observed followed by E. coli, with IBV infected came third in the row. On the bases of these findings we might conclude that mix infection of IBV and E. coli causes severe lesions with high morbidity and mortality. Availability: Items available for loan: UVAS Library [Call number: 2306-T] (1).
18.
Effects Of Omega 3 And Vitamin E Against Experimentally Infected Low Pathogenic Avian Influenza Virus H9n2 In Broiler Chickens
by Muhammad Sulman Ali Taseer (2008-VA-089) | Prof. Dr. Asim Aslam | Prof. Dr. Zafar Iqbal Chaudhry | Prof. Dr. Kamran Ashraf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Avian influenza is a highly contagious viral disease of domestic and wild birds. It is one of the most devastating viral disease of poultry industry. It was first identified in Italy in early 1900,s and is now known to exist worldwide. Total of 125 day old chickens were divided into five groups (A, B, C, D, E) with 25 chickens in each group. Group A was negative control group. In groups B, C, D and E low pathogenic avian influenza (H9N2) virus infection was introduced at day 28 of age. Group B was given with Omega 3. In group C chickens were given Vitamin. E. In group D chickens were fed both Omega 3 and Vitamin E. Group E was positive control group without any additional supplementation.
At days 27, 30, 35, 42, blood was collected aseptically from wing vein, from three birds in each group to check H/L ratio and to perform HI test to check antibody titer for H9. After collection of blood five birds from each group were slaughtered to observe postmortem signs and for the histopathology of lungs and trachea. Heterophill to lymphocyte ratio was significantly high in groups D (Omega 3 and Vit.E) and group E (Positive Control). Among the various treatment groups of broilers the significantly highest HI antibody titer was recorded in group E which was positive control group. In treatment groups C (Vitamin E supplement) and D (Omega 3 and Vit.E) HI antibody titer was near to protective titer against H9.
Major histopathological lesions involved deciliation of trachea and sloughing of epithelium of trachea. There was infiltration of monocytes and neutrophils as well as vascular congestion in the form of hemorrhagic areas in lungs. There was increase in congestion in the lungs of the chicks in group E (Positive Control).
37
FCR was evaluated on weekly basis. A comparatively better feed conversion ratio was recorded in group D (Omega 3 and Vit.E). There was no significant difference in feed conversion ratio of the other treatment groups. Availability: Items available for loan: UVAS Library [Call number: 2355-T] (1).
19.
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: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Theses submitted with cd. Availability: Items available for loan: UVAS Library [Call number: 2353-T] (1).
20.
Studies On The Effect Of Garvit-Pro® (A Commercial Product) On Broiler Chickens Vaccinated Against Infectious Bursal Disease
by Jawad Ahmad (2008-VA-109) | Prof. Dr. Asim Aslam | Dr. Muti Ur Rehman | Prof. Dr. Kamran Ashraf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Infectious Bursal Disease (IBD) has become very notorious poultry disease in Pakistan. The disease affects the primary lymphoid organs (bursa of Fabricius) of the poultry and cause mild to severe immune suppression in affected birds. In accordance of the mentioned fact, present experimental project was designed to evaluate the effects of immune boosters after IBD vaccination and compare the effects of GarVit-Pro with only vaccinated birds. Humoral immune response against IBD virus was measure through Enzyme Linked Immunosorbent Assay (ELISA). There were total 90 bird, divided into 3 groups with 30 birds in each group. The blood sample were collected from different experimental groups of broiler chickens on 07, 14, 21, 28 & 35 days of age. Among the various experimental groups of broiler chickens the significantly highest antibody titer was recorded in group B fed with GarVit-Pro (Garlic Supplement) as compared to the other vaccinated and unvaccinated groups. The histopathological scoring of Bursa of Fabricius and thymus in different experimental groups was observed at day 07, 14, 21, 28 and 35. The bursal lesion scores of Group B (GarVit-Pro Treated) was lower than the other two groups. The GarVit-Pro helped to enhance the antibody titer against IBD virus after vaccination. These findings suggest and advocates that GarVit-Pro (garlic supplement) can effectively stimulate and enhance the immunity in broiler chickens. GarVit-Pro can be potential ameliorator against different vaccines and their unwanted/suppressive effects in broiler chicks. It was proved that GarVit-Pro is able to implement immune response and have a patent immunomodulatory effect in chickens Availability: Items available for loan: UVAS Library [Call number: 2363-T] (1).
21.
Hematological And Histopathological Study Of Paramphistomum Cervi In Large Ruminants
by Abdul Majeed Saim (2012-va-814) | Prof. Dr. Asim Aslam | Mr. M. Saeed Imran | Prof. Dr. Kamran Ashraf .
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Paramphistomes are the parasites in the rumen, reticulum, duodenum and liver of goats, sheep, cattle and buffaloes. Their premature stages are in duodenum and then traveled towards the rumen. The damage caused by this illness in bovine affects production. These parasites aggravate a loss of weight, a lower nutritious conversion and a reduce in milk production. Lahore is one of the larger district in the Punjab province of Pakistan. The present study was conducted in order to implement efforts and regulation to eradicate the spread of disease in this area. From this study it was evaluated the changes in hematological parameters values of Large Ruminants and evaluated the Histopathological changes in rumen, reticulum, duodenum and liver.
Samples of blood and tissues were collected at slaughter house from cattle and buffaloes after slaughtering. These were divided into three groups i.e group-A (Infected Buffaloes=50), group-B (Infected Cattle=50) and group-C (Buffaloes, Cattle=20) serve as control. The blood samples and tissue samples of rumen, reticulum, duodenum and liver were taken from infected cattle and buffalo tested positive by fecal examination through direct smear technique and was processed for hematological and histopathological examination.
The infected cattle and buffaloes in this study described a highly significant decrease (p<0.05) in the mean red blood cells, packed cell volume, total erythrocyte count and hemoglobin. There was a significant increase (p<0.05) in the neutrophils number and eosinophil number of disease buffaloes and cows as compared to the non-infected buffaloes and cows. The infection of paramphistomum cervi is characterized by severe anemia, eosinophilia, neutrophilia. Anemia can be responsible for mortality in cattle and buffaloes especially young ones.
Pathological changes were observed grossly and confirmed histopathologically in the rumen, reticulum, duodenum and liver. Pathological changes were mostly limited to small intestine, especially duodenum. The disease caused by the parasite in the rumen was increased cornification of the stratum corneum, atrophy, severe infiltration and thickening of mucosa occurs in the rumen papillae, but no ulceration was found. Changes varied from a localized enteritis and villous atrophy in the duodenum in light disease to severe destructions of the mucosa extending into most of the jejunum in heavy infections. Mucosa at places was found congested with petechial haemorrhages. The histopathological analysis of the diseased liver of buffalos and cows has described that severe harm has occurred in this infection. In most of the sections, the normal structure of liver tissue, arrangement of hepatocyte cords, veins and portal tube areas were undergone clear with full bile ducts indicating bile duct hyperplasia, necrosis of liver tissue and cause necrosis. The immature forms of Paramphistomium cervi caused more severe damage in the duodenal tissue, where as adult forms inflicted mild tissue damage in the rumen.
The present study was conducted in order to implement efforts and regulation to eradicate the spread of disease in this area. From this study it was evaluated the changes in hematological parameters values of Large Ruminants and evaluated the Histopathological changes in rumen, reticulum, duodenum and liver.
Availability: Items available for loan: UVAS Library [Call number: 2368-T] (1).
22.
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: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Submitted with blank CD. Availability: Items available for loan: UVAS Library [Call number: 2369-T] (1).
23.
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: ; 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).
24.
Studies on Pathogenesis and Molecular Diagnosis of Infectious Bursal Disease Virus In Broiler Chicken
by Beenish Zahid (2003-VA-134) | Prof. Dr. Asim Aslam | Dr. Yasin Tipu | Prof. Dr. Tahir Yaqub.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Blank CD. Availability: Items available for loan: UVAS Library [Call number: 2510-T] (1).
25.
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).
26.
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
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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).
27.
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).
28.
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).
29.
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).
30.
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: ; 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
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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
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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).
31.
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).
32.
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).
33.
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).
34.
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).
35.
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).
36.
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: ; 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).
37.
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).
38.
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).
39.
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
Summary
71
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).
40.
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).
41.
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
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).
