Avian Influenza Virus Isolation, Propagation, and Titration in Embryonated Chicken Eggs

Author(s):  
Erica Spackman ◽  
Mary Lea Killian
2017 ◽  
Vol 91 (23) ◽  
Author(s):  
Mahmoud M. Naguib ◽  
Reiner Ulrich ◽  
Elisa Kasbohm ◽  
Christine L. P. Eng ◽  
Donata Hoffmann ◽  
...  

ABSTRACT The cocirculation of zoonotic highly pathogenic avian influenza virus (HPAIV) of subtype H5N1 and avian influenza virus (AIV) of subtype H9N2 among poultry in Egypt for at least 6 years should render that country a hypothetical hot spot for the emergence of reassortant, phenotypically altered viruses, yet no reassortants have been detected in Egypt. The present investigations proved that reassortants of the Egyptian H5N1 clade 2.2.1.2 virus and H9N2 virus of the G1-B lineage can be generated by coamplification in embryonated chicken eggs. Reassortants were restricted to the H5N1 subtype and acquired between two and all six of the internal segments of the H9N2 virus. Five selected plaque-purified reassortant clones expressed a broad phenotypic spectrum both in vitro and in vivo. Two groups of reassortants were characterized to have retarded growth characteristics in vitro compared to the H5N1 parent virus. One clone provoked reduced mortality in inoculated chickens, although the characteristics of a highly pathogenic phenotype were retained. Enhanced zoonotic properties were not predicted for any of these clones, and this prediction was confirmed by ferret inoculation experiments: neither the H5N1 parent virus nor two selected clones induced severe clinical symptoms or were transmitted to sentinel ferrets by contact. While the emergence of reassortants of Egyptian HPAIV of subtype H5N1 with internal gene segments of cocirculating H9N2 viruses is possible in principle, the spread of such viruses is expected to be governed by their fitness to outcompete the parental viruses in the field. The eventual spread of attenuated phenotypes, however, would negatively impact syndrome surveillance on poultry farms and might foster enzootic virus circulation. IMPORTANCE Despite almost 6 years of the continuous cocirculation of highly pathogenic avian influenza virus H5N1 and avian influenza virus H9N2 in poultry in Egypt, no reassortants of the two subtypes have been reported. Here, the principal compatibility of the two subtypes is shown by forcing the reassortment between copassaged H5N1 und H9N2 viruses in embryonated chicken eggs. The resulting reassortant viruses displayed a wide range of pathogenicity including attenuated phenotypes in chickens, but did not show enhanced zoonotic propensities in the ferret model.


2015 ◽  
Vol 28 (4) ◽  
pp. 192-200 ◽  
Author(s):  
Neda Barjesteh ◽  
Jennifer T. Brisbin ◽  
Shahriar Behboudi ◽  
Éva Nagy ◽  
Shayan Sharif

1985 ◽  
Vol 29 (4) ◽  
pp. 1195 ◽  
Author(s):  
D. T. Cappucci ◽  
D. C. Johnson ◽  
M. Brugh ◽  
T. M. Smith ◽  
C. F. Jackson ◽  
...  

2020 ◽  
Vol 11 (1) ◽  
pp. 1-6
Author(s):  
Amjad Ali Channa ◽  
Nazeer Hussain Kalhoro ◽  
Zaheer Ahmed Nizamani ◽  
Ayaz Hussain Mangi ◽  
Jamila Soomro

Background: Poultry is largest and rapidly growing sector of livestock in Pakistan. It is mainly influenced by viral pathogens such as Newcastle Disease Virus (NDV) and Avian Influenza Virus (H7N3). These viruses cause severe disease in poultry and leads to heavy economic losses throughout the world. The outbreaks of these pathogens have been increased in last few decades. Therefore, the study about antigenic prevalence is needed to know about the emergence of these pathogenic viruses, and to get rid of severe ailments associated with reduced poultry production. Objectives: To determine the prevalence of Newcastle Disease Virus (NDV), Avian Influenza Virus (H7N3) and co-infections in poultry flocks at Karachi. Methodology: For detection of NDV and H7N3, a total of 200 tracheal swabs were collected and tested through virus isolation (V.I); the sample with positive virus isolation were tested through agar gel precipitation (AGP) and then the RNA was isolated through TRI Reagent, which was further tested through reverse transcription polymerase chain reaction (RT-PCR). Results: The virus isolation showed that 58% of samples were positive for various viruses. Agar gel precipitation (AGP) revealed that the occurrence of NDV, H7N3 and ND+H7 were 50%, 8% and 38%, respectively. RT-PCR for F and HA gene of NDV and H7N3 confirmed the presence of NDV and H7N3 in the poultry. Conclusion: It is concluded that NDV and H7N3 are circulating in the flocks causing co-infections, therefore it is important to know the field challenge of viruses and to prepare vaccine of circulating serotype of virus to mitigate the rate of infection.


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