Isolation and characterization of highly pathogenic avian influenza virus subtype H5N1 from donkeys.
BACKGROUND
The highly pathogenic H5N1 is a major avian pathogen that crosses species barriers and seriously affects humans as well as some mammals. It mutates in an intensified manner and is considered a potential candidate for the possible next pandemic with all the catastrophic consequences.


METHODS
Nasal swabs were collected from donkeys suffered from respiratory distress. The virus was isolated from the pooled nasal swabs in specific pathogen free embryonated chicken eggs (SPF-ECE). Reverse transcriptase polymerase chain reaction (RT-PCR) and sequencing of both haemagglutingin and neuraminidase were performed. H5 seroconversion was screened using haemagglutination inhibition (HI) assay on 105 donkey serum samples.


RESULTS
We demonstrated that H5N1 jumped from poultry to another mammalian host; donkeys. Phylogenetic analysis showed that the virus clustered within the lineage of H5N1 from Egypt, closely related to 2009 isolates. It harboured few genetic changes compared to the closely related viruses from avian and humans. The neuraminidase lacks oseltamivir resistant mutations. Interestingly, HI screening for antibodies to H5 haemagglutinins in donkeys revealed high exposure rate.


CONCLUSIONS
These findings extend the host range of the H5N1 influenza virus, possess implications for influenza virus epidemiology and highlight the need for the systematic surveillance of H5N1 in animals in the vicinity of backyard poultry units especially in endemic areas.
Abdel-Moneim
Ahmed S
AS
Department of Virology, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef 62511, Egypt. asa@bsu.edu.eg
Abdel-Ghany
Ahmad E
AE
Shany
Salama A S
SA
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D001483:Base Sequence; D004534:Egypt / Q000453:epidemiology; D018397:Equidae / Q000821:virology*; D005814:Genes, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification* / Q000472:pathogenicity; D017720:Molecular Epidemiology; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000235:genetics; D009976:Orthomyxoviridae Infections / Q000453:epidemiology / Q000635:transmission / Q000662:veterinary* / Q000821:virology; D010802:Phylogeny; D011200:Poultry / Q000821:virology; D012367:RNA, Viral / Q000235:genetics; D012141:Respiratory Tract Infections / Q000453:epidemiology / Q000635:transmission / Q000662:veterinary / Q000821:virology; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D012367:RNA, Viral; D009439:Neuraminidase
J Virol. 2005 Mar;79(5):2814-22
15709000
Virology. 1994 Nov 15;205(1):17-23
7975212
Virology. 1992 May;188(1):245-55
1314452
Science. 2004 Oct 8;306(5694):241
15345779
Science. 2006 Apr 21;312(5772):404-10
16543414
Emerg Infect Dis. 2006 Nov;12(11):1744-7
17283627
Virology. 1989 Nov;173(1):317-22
2815586
J Virol. 2000 Nov;74(22):10807-10
11044127
Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2845-50
16473931
Nature. 2006 Nov 16;444(7117):378-82
17108965
Bioinformatics. 2001 Dec;17(12):1244-5
11751241
Virology. 2006 Jan 20;344(2):432-8
16226289
J Virol. 1999 Apr;73(4):3184-9
10074171
J Virol. 2002 May;76(9):4420-9
11932409
J Virol. 1999 Feb;73(2):1146-55
9882316
J Virol. 2005 Aug;79(15):9926-32
16014953
Virology. 1983 Jun;127(2):361-73
6868370
Intervirology. 1993;35(1-4):16-25
8407243
J Comp Pathol. 2007 Aug-Oct;137(2-3):155-9
17689552
Emerg Infect Dis. 2008 Jul;14(7):1132-4
18598640
Emerg Infect Dis. 2004 Dec;10(12):2189-91
15663858
J Virol. 2007 Nov;81(22):12418-26
17855542
Nucleic Acids Res. 2007 Jul;35(Web Server issue):W275-9
17537820
J Virol. 2007 Sep;81(18):9950-5
17626098
Am J Respir Crit Care Med. 1995 Oct;152(4 Pt 2):S16-9
7551406
Comp Immunol Microbiol Infect Dis. 2009 Jul;32(4):255-73
18485480
Vet Pathol. 2003 May;40(3):304-10
12724572
BMC Struct Biol. 2009 Apr 07;9:21
19351406
J Virol. 2000 Dec;74(24):11825-31
11090182
J Mol Biol. 1990 Oct 5;215(3):403-10
2231712