I= MOLECULAR CHARACTERIZATION OF PAKISTANI ISOLATES OF HYDROPERICARDIUM SYNDROME AND PRODUCTION OF ITS VACCINE
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Title of Thesis
MOLECULAR CHARACTERIZATION OF PAKISTANI ISOLATES OF HYDROPERICARDIUM SYNDROME AND PRODUCTION OF ITS VACCINE
Author(s)
SEEMA SHAMIM
Institute/University/Department Details
Department of Physiology/ University of Karachi
Session
2005
Subject
Physiology
Number of Pages
165
Keywords (Extracted from title, table of contents and abstract of thesis)
hydropericardium syndrome, vaccine, poultry, hemagglutination, agar gel precipitation, serum neutralization, avian adeno virus

Abstract
Hydropericardium syndrome (HPS), a disease of chickens, has drastically affected the poultry industry in Pakistan and needs an effective therapeutic approach like vaccination. Understanding of the causative agent is essential for such an approach. Various characteristic features of the causative agent and its molecular characterizations was carried out during the present investigation.

The passage has been made in susceptible chickens and the isolates collected during successive periods (1990-2003) were used. These isolates were characterized by experimental serological tests (agar gel precipitation, serum neutralization and indirect hemagglutination), propagation in various systems (chicken LD 50, intravenous pathogenicity index-and intracerebal pathogenicity index) and preparation of live (freeze-dried) and in-activated (using formalin and oil emulsion) vaccine was accomplished and documented by standard microbiological routine tests like sterility, potency and sensitivity tests. Finally, extraction of DNA form selected HPS isolates along with polymerase chain reaction (PCR), Restriction Fragment Length Polymorphism (RFLP), followed by agarose gel electrophoresis were carried out. There no differences in size of all the PCR products for H3 /H4 primer pair (approximately 1319 bp), which is characteristic of group 1 avian adenovirus. Restriction fragment length polymorphism of these PCR products by HPA restriction enzyme digestion yielded five bands approximately 0.15,0.2,0.25,0.3 and 0.5 Kbp. These profiles are identical to those of avian adenovirus isolates reported from India, Germany, Japan and Chile. Detection of small amount of DNA enabled the viral DNA to amplified directly from field isolates and avoid and avoid an extra seep to grow the virus in cell culture and the laborious work in serology. The present findings document that these Pakistani isolates belong to Avian adenovirus group I serotypes 4 (FAV4).
Download Full Thesis
3094.11 KB
S. No. Chapter Title of the Chapters Page Size (KB)
1 0 Contents
161.65 KB
2 1 General Introduction
130.11 KB
  1.1 General Introduction 2
  1.2 Hypothesis And Objectives 12
3 2 General Review Of Literature
226.7 KB
  2.1 Disease 14
  2.2 Mortality 14
  2.3 Pathology Features 16
  2.4. Histopathology 18
  2.5 Epidemiology 20
  2.6 Physico -Chemical Characterization 21
  2.7 Etiology 21
  2.8 Infection 26
  2.9 Avian Adeno Virus Associated With HPS 31
  2.10 Molecular Etiology 32
4 3 General Materials And Methods
128.35 KB
  3.1 Materials And Methods 34
  3.2 Experimental Chickens 34
  3.3 Sterilization 35
  3.4 Lips-Virus (Lips I Spvc ) 35
  3.5 Production Of Antisera 36
  3.6 Serological Test 36
  3.7 Sero Conversion In Chickens 38
  3.8 Serum Neutralization Test 38
  3.9 Indirect Haemagglutination Test 39
5 4 Propagation Of HPS Virus
1769.04 KB
  4.1 Introduction 50
  4.2 Materials And Methods 50
  4.3 Results And Discussion 53
6 5 P A Thogencity Of Bps Virus
128.97 KB
  5.1 Introduction 67
  5.2 General Observation Regarding Factors Affecting The Pathogenicity /Viability Of Hps Virus 67
7 6 Preparation Of Bps Vaccines And Their Comparison
103.42 KB
  6.1 Introduction 84
  6.2 Materials And Methods 84
  6.3 Preparation Of Vaccine By Using Susceptible Chicken 91
  6.4 Results And Discussion 93
8 7 Molecular Characterization Of Bps Virus
296.35 KB
  7.1 Introduction 98
  7.2 Materials And Methods 99
  7.3 Detection Of Genomic Nucleic Acids 101
  7.4 Restriction Fragment Length Polymorphism 109
  7.5 Results And Discussion 110
9 8 General Discussion And Conclusions
377.3 KB
  8.1 General Discussion 126
  8.2 Conclusions And Prospective Future Plans 137
  8.3 References 128