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تحديد القرابة الوراثية لعزلات عراقية ضارية لفيروس مرض النيوكاســــل من تتابع القواعد النتروجينية في موروث الهموكلوتنين نيورامنيديز == Phylogenetic Determination of Newcastle Disease Virus Hn Gene Sequences Iraqi Virulent Isolate
Author name:
مرتضى عبد المهدي محمد حسن المظفر
Supervisor name:
هدى عبد الهادي علي النصراوي | احمد مجيد حمزة الشمري
General topic:
Veterinary Medicine
Specific topic:
Common Diseases
Degree:
Master
University:
University of Al-Qadisiyah - College Of Veterinary Medicine
Language:
English
University location:
Qadisiyah
First pages:
21T494 - p.pdf
Abstract:
يعد مرض النيوكاسل من اخطر الامراض الفيروسية المعدية التي تصيب قطعان الدواجن وتسبب خسائر اقتصادية كبيرة في صناعة الدواجن واول تسجيل لمرض النيوكاسل كمرض مشترك بين الانسان والحيوان كان من قبل العالمBurnet في سنة 1943. والهدف من الدراسة هو عزل وتحليل تسل | Newcastle disease is considered to be the most contagious poultry disease and may cause severe economic loss in the poultry industry. The first report in which Newcastle disease virus (NDV) was consider as zoonotic disease and a human pathogen that was published by Burnet, in 1943. The aim of this study was to isolate and determination of nucleotide sequence of the Hemagglutinin - neuraminidase gene (HN) of two NDV Iraqi virulent isolates. These isolates were named as Najaf isolate (Najaf APMV1/ Chicken/ Iraq - Najaf/ ICCMGR/2012) and Baghdad isolate (Baghdad ICCMGR). This work was conducted in Iraq for the first time, the reason behind it was to correlate phylogenetically between these two isolates and the NDV strains in the countries of the region. This will help in determining the source of NDV outbreaks occurred in Iraq. Both Najaf and Baghdad isolates were propagated in embryonated chicken eggs, after inoculation of 11 days embryonated chicken eggs was showed and embryos were killed in different times (more than 40 hours and less than 72 hours). To assess the activity of both virus isolates, the main features of the infected embryos that reflect characteristics of congestion and sever hemorrhage was compared with control one. Hemorrhage was markedly severed in the infected embryos and it was similar in its severity by both isolates. The allantoic fluid was harvested post infection and the agglutination activity reflected titer of 1024 for Najaf isolate and 512 for Baghdad isolate. In order to isolate the Hemagglutinin - neuraminidase gene (HN) from both Najaf and Baghdad NDV isolates, specific primers for this gene were designed. After viral RNA purification, one - step reverse transcriptase - PCR was undertaken to amplify the NH gene and isolated it from the gel. Nucleotide sequence of the isolated NH gene from both virus isolates was determined. The purpose was to focus on molecular and phylogenetic analysis of the hemagglutinin - neuraminidase (HN) gene in both Iraqi isolates. The resulted nucleotide sequence of 647 nucleotides of the HN gene for both isolate were submitted to the GenBank database under the accession numbers KJ632972. To determine the origin of both Iraqi virulent isolates, HN gene sequence of both isolates was aligned with sequences of NDV isolates previously published in GenBank. The resulted alignment was then analyzed in terms of phylogenetic relation and variation. phylogenetic analysis of the isolated virus showed that both Iraqi isolates share significant similarity with 14 other international stains. Two strains of India origin and 12 was China isolates. The identity percentage was 99% - 100% with clustered group. Interestingly, Iraqi isolate was different from the neighbor countries such as Iran, and other Middle East countries. These results may indicate that certain migratory birds might have contributed to the distribution of NDV in Iraq. However imported infected beards from south East Asia may contribute to the current NDV outbreaks caused by foreign strain.
