Cell Host & Microbe

Article

Circulating Avian Influenza Viruses

Closely Related to the 1918 Virus
Have Pandemic Potential
Tokiko Watanabe,1,2,12 Gongxun Zhong,1,12 Colin A. Russell,3,12 Noriko Nakajima,4 Masato Hatta,1 Anthony Hanson,1
Ryan McBride,5 David F. Burke,6 Kenta Takahashi,4 Satoshi Fukuyama,2 Yuriko Tomita,2 Eileen A. Maher,1
Shinji Watanabe,2,7 Masaki Imai,8,9 Gabriele Neumann,1 Hideki Hasegawa,4 James C. Paulson,5 Derek J. Smith,6
and Yoshihiro Kawaoka1,2,10,11,*
1Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 575 Science Drive, Madison,

WI 53711, USA
2ERATO Infection-Induced Host Responses Project, Japan Science and Technology Agency, Saitama 332-0012, Japan
3Department of Veterinary Medicine, University of Cambridge, Madingley Road, Cambridge CB3 0ES, UK
4Department of Pathology, National Institute of Infectious Diseases, Shinjuku, Tokyo 162-8640, Japan
5Department of Cell and Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA
6Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ, UK
7Laboratory of Veterinary Microbiology, Department of Veterinary Sciences, University of Miyazaki, Miyazaki 889-2192, Japan
8Department of Veterinary Medicine, Faculty of Agriculture, Iwate University, Iwate 020-8550, Japan
9Influenza Virus Research Center, National Institute of Infectious Diseases, Tokyo 208-0011, Japan
10Division of Virology, Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639,

Japan
11Department of Special Pathogens, International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo,

Minato-ku, Tokyo 108-8639, Japan

12Co-first authors

*Correspondence: kawaokay@[Link]
[Link]

SUMMARY infectious diseases. Most of these so-called emerging infec-

tious diseases are caused by zoonotic pathogens that originate
Wild birds harbor a large gene pool of influenza A vi- in animals (Jones et al., 2008; Taylor et al., 2001). These path-
ruses that have the potential to cause influenza pan- ogens are responsible for various human diseases, such as
demics. Foreseeing and understanding this potential AIDS, SARS, and pandemic influenza. Animal-origin pathogens
is important for effective surveillance. Our phyloge- must overcome an immense hurdle to cause a zoonosis (i.e.,
netic and geographic analyses revealed the global interspecies transmission from natural or intermediate hosts
to humans). If such pathogens acquire the ability to transmit
prevalence of avian influenza virus genes whose
among humans, they become an appreciable threat to
proteins differ only a few amino acids from the 1918
mankind.
pandemic influenza virus, suggesting that 1918- The 1918 influenza pandemic, the most devastating dis-
like pandemic viruses may emerge in the future. To ease outbreak recorded, killed an estimated 40–50 million
assess this risk, we generated and characterized a vi- people worldwide (Johnson and Mueller, 2002; Taubenberger
rus composed of avian influenza viral segments with and Morens, 2006). Sequence analyses identified the 1918
high homology to the 1918 virus. This virus exhibited pandemic strain as an H1N1 influenza A virus of avian origin,
pathogenicity in mice and ferrets higher than that in although this conclusion remains controversial (Rabadan
an authentic avian influenza virus. Further, acquisi- et al., 2006; Reid et al., 2004; Smith et al., 2009). Since avian
tion of seven amino acid substitutions in the viral species harbor a large influenza virus gene pool that may
polymerases and the hemagglutinin surface glyco- contain influenza viral segments encoding proteins with high
homology to the 1918 viral proteins (designated as 1918
protein conferred respiratory droplet transmission
virus-like virus proteins), the possibility exists for a 1918 vi-
to the 1918-like avian virus in ferrets, demonstrating
rus-like avian virus to emerge in the human population as a
that contemporary avian influenza viruses with 1918 pandemic virus; however, the likelihood of such an event re-
virus-like proteins may have pandemic potential. mains unknown. To assess the risk of emergence of pandemic
influenza viruses reminiscent of the 1918 influenza virus, we
examined the properties of influenza viruses composed
INTRODUCTION of avian influenza viral segments that encode proteins with
high homology to the 1918 viral proteins (designated as
Despite having conquered many infectious diseases, we 1918-like avian viruses), which we generated by using reverse
continue to face a threat from novel, previously unrecognized genetics.

692 Cell Host & Microbe 15, 692–705, June 11, 2014 ª2014 Elsevier Inc.
Cell Host & Microbe
Pandemic Potential of Avian Influenza Virus

RESULTS growth was comparable to that of the 1918 virus (8.8 ± 0.1 and
8.4 ± 0.2 log10 pfu/ml in MDCK cells and eggs, respectively, at
Generation of a 1918-like Avian Virus Possessing Avian 24 hr postinfection).
Influenza Viral Segments that Encode Proteins with High
Homology to the 1918 Viral Proteins The 1918-like Avian Virus Exhibits Intermediate
We first determined whether influenza A virus gene segments Pathogenicity Compared with the 1918 Virus and an
that encode proteins with high homology to the 1918 viral pro- Authentic Avian Virus in Mouse and Ferret Models
teins exist in the avian influenza virus gene pool. We focused To assess the pathogenicity of the 1918-like avian virus, we first
on amino acid sequence comparisons because our goal was determined the mouse lethal dose 50 (MLD50; the dose required
to identify avian influenza viral proteins that closely resemble to kill 50% of infected mice) values of the 1918-like avian influ-
1918 virus proteins in structure and function and may therefore enza virus and authentic 1918 virus. As a control for the authentic
allow the emergence of a 1918-like virus. To this end, we per- avian influenza virus, we used A/duck/ALB/35/76 (H1N1;
formed a sequence similarity search using BLAST ([Link] DK/ALB) because this avian strain was well characterized in a
[Link]/[Link]) to identify the closest relatives to previous study (Van Hoeven et al., 2009). The 1918-like avian
the 1918 viral proteins. Interestingly, for most viral proteins influenza virus showed intermediate pathogenicity (with an
(except for hemagglutinin [HA], neuraminidase [NA], and PB1- MLD50 of 5.5 log10 pfu) compared with DK/ALB (with an MLD50
F2), we found avian influenza virus proteins that differed from of 6.8 log10 pfu) and the 1918 virus (with an MLD50 of 2.7 log10
their 1918 counterparts by only a limited number of amino acids pfu). These data indicate that the avian influenza virus genes
(Table S1). For example, for polymerase basis 2 (PB2) we found that were selected because of their close relationship with
one avian influenza PB2 protein that differed from 1918 PB2 by 1918 virus proteins not only cooperate at a functional level, but
eight amino acids. Similarly, we found avian influenza PB1, poly- also support pathogenicity higher than that of an authentic avian
merase acid (PA), nucleoprotein (NP), matrix protein 1 (M1), M2, influenza virus.
nonstructural protein 1 (NS1), and NS2 proteins that closely The ferret is considered the best current model for influenza
resembled their 1918 counterparts. Most of the viruses from virus infection because infected animals exhibit symptoms
which these proteins were derived were isolated recently, sug- that resemble those of humans infected with influenza A virus.
gesting that 95 years after the devastating 1918 pandemic, avian Therefore, we next tested the pathogenicity of the 1918-like
influenza virus genes encoding 1918-like proteins continue to avian virus in ferrets. Ferrets (three animals per group) were
circulate in nature. For the 1918 HA and NA proteins, we identi- intranasally inoculated with 106 pfu/500 ml of the 1918-like avian
fied the closest avian H1 and N1 relatives, respectively. These virus, 1918 virus, or DK/ALB virus (Figure S1, available online).
proteins differed from the 1918 proteins by 33 and 31–33 amino All animals infected with the 1918 virus became symptomatic;
acids, respectively, a finding that was expected due to the higher their body weights declined drastically (the mean maximum
evolutionary rates in these genes relative to the other influenza body weight loss was 17.5% ± 2.2%), and one of them died
viral genes. on day 8 postinfection (Figure S1A). By contrast, none of the
To assess the risk of emergence of a 1918-like virus and to ferrets infected with DK/ALB exhibited noticeable clinical signs
delineate the amino acid changes that are needed for such a (no appreciable body weight loss in any of the animals; Fig-
virus to become transmissible via respiratory droplets in mam- ure S1C), whereas animals infected with the 1918-like avian vi-
mals, we attempted to generate an influenza virus composed rus became symptomatic and showed substantial body weight
of avian influenza viral segments that encoded proteins with loss (the mean maximum body weight loss was 11.9% ± 3.9%;
high homology to the 1918 viral proteins. In particular, we see Table 1 and Figure S1B). Of the three viruses tested, the
selected the following genes to generate a 1918-like avian 1918 virus replicated the most efficiently in the upper and lower
influenza virus (referred to as 1918-like avian virus) (see Supple- respiratory tract (p < 0.01; Figure 1 and Table S2). The 1918-
mental Experimental Procedures): the PB2 segment of A/blue- and 1918-like avian virus-infected ferrets displayed numerous
winged teal/Ohio/926/2002 (H3N8), the PB1 segment of viral antigen-positive cells in tracheal, bronchial, bronchiolar,
A/blue-winged teal/Alberta (ALB)/286/77 (H3N6), the PA and glandular epithelial cells and necrotized changes in some
segment of A/pintail duck/ALB/219/77 (H1N1), the NP segment trachea-bronchial glands on day 6 postinfection (Figure 2A),
of A/blue-winged teal/Ohio/908/2002 (H1N1), the M segment whereas the 1918-like avian virus-infected ferrets presented
of A/duck/Germany/113/95 (H9N2), the NS segment of few antigen-positive cells on day 3 postinfection (Figure S2).
A/canvasback duck/Alberta/102/76 (H3N6), the HA segment of In the lungs, no significant differences in pathologic changes
A/pintail duck/ALB/238/79 (H1N1), and the NA segment were detected between the 1918 and 1918-like avian virus
of A/mallard/duck/ALB/46/77 (H1N1), as described in the Sup- groups (Figures 2A, 2B, and S2). In contrast, the DK/ALB infec-
plemental Information. The resulting virus differs by 8 (PB2), 6 tion caused only minimal to mild bronchitis or bronchiolitis with
(PB1), 20 (PB1-F2), 9 (PA), 7 (NP), 33 (HA), 31 (NA), 1 (M1), 5 little viral antigen expression on days 3 and 6 postinfection (Fig-
(M2), 4 (NS1), and 0 (NS2) amino acids from the 1918 virus. ures 2A, 2B, and S2). Taken together with the results of recent
The 1918-like avian virus possessing these eight 1918-like studies that showed no appreciable clinical signs in ferrets
avian viral segments was successfully recovered from 293T cells infected with more commonplace avian influenza viruses of
transfected with the plasmids required to generate this virus. The various subtypes (Koçer et al., 2012; Watanabe et al., 2013),
virus grew well in Madin-Darby canine kidney (MDCK) cells and the finding that the 1918-like avian influenza virus is of interme-
embryonated chicken eggs (8.3 ± 0.1 and 8.6 ± 0.2 log10 plaque- diate pathogenicity in mammals may suggest that the progeni-
forming units (pfu)/ml at 24 hr postinfection, respectively), and its tor of the 1918 virus was an unusual avian influenza virus whose

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Table 1. Pathogenicity and Transmissibility of Reassortants and Mutants of 1918-like Avian Virus in Ferrets
Virus-Inoculated Ferrets Contact Ferrets
Survival (No. of Virus Detection in Nasal Wash Seroconversion (Positive
Virus Weight Loss (%)a Dead/Total) (Positive and Total Numbers) and Total Numbers)b
1918 17.5 ± 2.2 1/3 2 of 3 2 of 3
1918-like Avianc 11.9 ± 3.9 0/3 0 of 3 0 of 3
DK/ALB 1.9 ± 3.3 0/3 0 of 3 0 of 3
1918 PB2/Avian 17.8 ± 2.2 0/3 0 of 3 0 of 3
1918 HA/Avian 16.9 ± 5.8 2/3 0 of 3 0 of 3
1918 PB2:HA/Avian 18.7 ± 4.0 2/3 0 of 3 0 of 3
1918 PB2:HA:NA/Avian 18.6 ± 3.7 0/3 2 of 3 2 of 3
1918(3P+NP):HA/Avian 21.2 ± 4.4 2/3 1 of 3 1 of 3
1918-like Avian 10.3 ± 3.6 0/3 0 of 3 0 of 3
PB2-627K
1918-like Avian 19.4 ± 9.2 1/3 1 of 3 1 of 3
PB2-627K
HA-89ED/190D/225D
1918-like Avian 21.0 ± 5.1 0/3 2 of 3 2 of 3
PB2-627K/684D:HA-89ED/113SN/
190D/225D/265DV:PA-253M
See also Figures S1 and S3 and Table S3. For each pair of ferrets, one animal was intranasally inoculated with 106 pfu of virus (0.5 ml) (virus-inoculated
ferret), and 1 day later a naive ferret was placed in an adjacent cage (contact ferret). Virus-inoculated ferrets were monitored for 14 days to determine
survival rates and body weight changes.
a
Maximum percentage weight loss (mean ± SD) is shown.
b
Serum samples were collected 18 days after infection; HI assays were carried out with homologous virus and turkey red blood cells.
c
The 1918-like avian virus possessing the avian influenza viral segments that encoded proteins with high homology to the 1918 viral proteins described
in the Supplemental Experimental Procedures was generated by using reverse genetics. The effectiveness of current vaccines and antiviral drugs
against the 1918-like avian virus was examined (see Tables S4 and S5).

pathogenicity in mammals was higher than that of most avian or the 1918 HA gene were higher than that of the 1918-like avian
influenza viruses. virus on day 3 postinfection (Table S2), although these differ-
ences were not statistically significant. These results suggest
The 1918 PB2 and HA Genes Contribute to Enhanced that the 1918 PB2 and HA genes confer high pathogenicity to
Pathogenicity and Transmissibility in Ferrets the 1918-like avian virus in the ferret model.
To identify the 1918 viral segments that are responsible for the For an influenza virus to cause a pandemic, it must achieve
intermediate pathogenicity of the 1918-like avian virus relative efficient human-to-human transmission. Therefore, we tested
to the 1918 and authentic avian viruses, we examined the path- transmissibility in ferrets of the authentic 1918 virus, as well as
ogenicity of reassortant viruses possessing 1918 viral genes in the 1918-like avian virus and its reassortants. Three animals
the genetic background of the 1918-like avian virus in ferrets. were each intranasally inoculated with 106 pfu of virus. On day
We focused on the HA and PB2 genes, which are known to 1 after infection, a naive ferret was housed in a cage adjacent
play important roles in the adaptation of avian influenza viruses to each of the infected ferrets. This setup prevented direct con-
to mammals (Hatta et al., 2001; Li et al., 2005; Matrosovich tact between animals but allowed the spread of influenza virus
et al., 2008; Neumann and Kawaoka, 2006; Subbarao et al., through respiratory droplets. Viral titers were determined in nasal
1993). We generated 1918 PB2/Avian, 1918 HA/Avian, and washes collected from both the inoculated and contact ferrets
1918 PB2:HA/Avian viruses, which possess the PB2, HA, or every other day postinfection/contact (for up to 9 days). The
PB2 and HA genes of the 1918 virus, respectively, and the 1918 virus was recovered from two of the three contact ferrets
remaining genes from the 1918-like avian virus (Figure S3). We (Figure 3A), demonstrating respiratory droplet transmission of
also created 1918 PB2:HA:NA/Avian and 1918(3P+NP):HA/ the 1918 virus. In contrast, the 1918-like avian and DK/ALB
Avian viruses, which possess the 1918 virus PB2, HA, and NA viruses failed to transmit among ferrets; no virus was detected
genes, or the 1918 virus PA, PB1, PB2, NP, and HA genes, in nasal washes collected from contact animals, although the
respectively, and the remaining genes from the 1918-like avian inoculated animals did shed virus (Figures 3B and 3C). Similarly,
virus (Figure S3). All animals infected with these viruses became no virus was recovered from contact animals for the 1918 PB2/
symptomatic; they lost appetite and body weight (Table 1 and Avian, 1918 HA/Avian, and 1918 PB2:HA/Avian virus groups
Figure S1). Two of three ferrets died upon infection with the (Figures S4A–S4C). However, virus was recovered from one of
1918 HA/Avian, 1918 PB2:HA/Avian, and 1918(3P+NP):HA/ the three contact animals in the 1918(3P+NP):HA/Avian and
Avian viruses (Table 1). In the ferret tracheae, the mean virus two of three contact ferrets in the 1918 PB2:HA:NA/Avian virus
titers of all reassortant viruses possessing the 1918 PB2 gene group (Figures S4D and S4E), suggesting potential roles for the

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Figure 1. Replicative Ability of 1918 Virus,

an Authentic Avian Virus, 1918-like Avian
Virus, and 1918-like Avian Mutant Viruses
Virus replication in respiratory organs of ferrets.
Ferrets were intranasally infected with 106 pfu of
virus. Six animals per group were euthanized on
days 3 and 6 postinfection for virus titration. Virus
titers in nasal turbinates, tracheae, and lungs were
determined by plaque assay in MDCK cells. Hori-
zontal bars indicate the mean virus titers. Asterisks
indicate virus titers significantly different from
those of the 1918-like avian virus (*p < 0.05; **p <
0.01). See also Table S2.

but we were able to obtain this virus after

propagation of 293T cell supernatant
in MDCK cells. However, this virus con-
sisted of a mixed virus population pos-
sessing E or D at position 89 of HA
(89ED) in addition to the PB2-627K and
HA-190D/225D mutations (designated
as 1918-like avian PB2-627K:HA-89ED/
190D/225D; Figure S3 and Table 2).
These mutant viruses replicated well
in the nasal turbinates and tracheal tis-
sues of ferrets (Figure 1 and Table S2).
Infection of ferrets with 1918-like avian
PB2-627K and 1918-like avian PB2-
627K:HA-89ED/190D/225D caused sub-
RNA replication complex, HA, and NA in virus transmission stantial body weight loss (10.3% ± 3.6% and 19.4% ± 9.2%,
among ferrets. Taken together, our data suggest that the 1918 respectively; Table 1) and appreciable pathologic changes in
PB2 and HA genes confer enhanced pathogenicity and trans- the trachea and lungs of the infected animals (Figures 2A and
missibility to the 1918-like avian virus in ferrets. S2). We tested the transmissibility of these viruses and found
that no virus was recovered from contact ferrets for the 1918-
Identification of Amino Acid Substitutions Potentially like avian PB2-627K virus group. Interestingly, for the 1918-like
Associated with the Efficient Transmission of the avian PB2-627K:HA-89ED/190D/225D virus group, virus and
1918-like Avian Virus in Ferrets seroconversion were detected for one of the three contact fer-
HA and PB2 are known to play important roles in restricting viral rets (Table 1, Figures 3E and S4F, and Table S3), indicating its
transmission from avian species to humans (Van Hoeven et al., partial transmissibility in this animal model.
2009). The receptor-binding specificity of the HA protein is a ma- We sequenced the virus isolated on days 5 and 9 postcontact
jor determinant of influenza viral host range (Matrosovich et al., from the nasal washes of the contact ferret in the 1918-like avian
2008). E-to-D and G-to-D mutations at positions 190 and 225 PB2-627K:HA-89ED/190D/225D virus group and found three
of HA (H3 numbering), respectively, are essential for avian virus additional mutations, HA-S113N, PB2-A684D, and PA-V253M
HAs of the H1 subtype to bind to human-type receptors (Matro- (Table 2). After propagating this virus in MDCK cells, the virus
sovich et al., 2000). In addition, Lys at position 627 of the PB2 stock possessed the following mutations: PB2-627K, PB2-
protein is important for avian viruses to efficiently replicate in 684D, PA-253M, HA-190D, HA-225D, HA-89E/D, HA-113S/N,
mammalian cells and at the lower temperatures of the upper and HA-265D/V (the mixed populations of amino acids were found
human airway (Hatta et al., 2001, 2007; Subbarao et al., 1993). at positions 89, 113, and 265) (designated as 1918-like avian
To test whether these mammalian-adapting mutations in HA PB2-627K/684D:HA-89ED/113SN/190D/225D/265DV:PA-253M)
and PB2 affect the replicative ability, pathogenicity, and trans- (Table 2 and Figure S3).
missibility of the 1918-like avian virus, we attempted to generate The 1918-like avian PB2-627K/684D:HA-89ED/113SN/190D/
mutant 1918-like reassortant avian viruses possessing these 225D /265DV:PA-253M virus replicated in trachea and lungs
amino acid substitutions (i.e., 1918-like avian PB2-627K and more efficiently than the 1918-like avian virus (p < 0.05; Figure 1
1918-like avian PB2-627K:HA-190D/225D viruses). 1918-like and Table S2) and caused severe weight loss in the infected fer-
avian PB2-627K virus was generated upon inoculation of the rets (maximum body weight loss was 21.0% ± 5.1%), although no
supernatant of transfected 293T cells into embryonated chicken infected animals died (Table 1). The 1918-like avian PB2-627K/
eggs (Figure S3). We were unable to generate 1918-like avian 684D:HA-89ED/113SN/190D/225D/265DV:PA-253M infection
PB2-627K:HA-190D/225D virus in embryonated chicken eggs, caused more progressive inflammation in the lungs of ferrets on

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Figure 2. Pathological Analyses of 1918 Virus, an Authentic Avian Virus, 1918-like Avian Virus, and 1918-like Avian Mutant Viruses
(A) Histopathological findings in virus-infected ferrets. Shown are representative pathological changes in tracheae and lungs of ferrets infected with 106 pfu of the
indicated viruses on day 6 postinfection. Three ferrets per group were infected intranasally with 106 pfu of virus, and tissues were collected on day 6 after infection
for pathological examination. No virus was detected from the lungs of the DK/ALB-infected ferrets. Left: H&E staining. Right: immunohistochemical staining for
influenza viral antigen (NP). Scale bars, 100 mm.
(B) Pathological severity scores for infected ferrets. To represent comprehensive histological changes, respiratory tissue slides were evaluated by scoring
pathological changes as described in the Supplemental Experimental Procedures. The sum of the pathologic scores for all five lung lobes was calculated for each
ferret. The means ± SD from three ferrets are shown. Asterisks indicate virus pathological scores significantly different from that of the 1918-like avian virus
(Dunnett’s test; p < 0.05). See also Figure S2.

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day 3 postinfection compared with that caused by the 1918-like 190D/225D (Figures 4 and S5); however, the biological signifi-
avian virus (Dunnett’s test; p < 0.05; Figures 2B and S2). The cance is currently unknown.
ferrets infected with this mutant virus presented numerous In addition to receptor binding, another critical function of HA
viral antigen-positive cells in tracheal, bronchial, bronchiolar, is membrane fusion. A recent study showed a difference in the
and glandular epithelial cells on days 3 and 6 postinfections (Fig- pH at which HA is activated for fusion between transmissible
ures 2A and S2). We then tested the transmissibility of this virus and nontransmissible viruses (de Vries et al., 2014; Imai et al.,
and found that two of the three contact ferrets were positive for 2012). Therefore, we next examined HA fusion activity under
virus between days 5 and 9 after contact; these animals were different pH conditions by testing the polykaryon formation
also seropositive (Figures 3F and S4F and Table S3). Sequence efficiency of HeLa cells expressing 1918 HA, the 1918-like
analysis showed that the virus recovered from the contact animal avian HA, the 1918-like avian mutant HAs, or DK/ALB HA (Fig-
for pair 1 possessed an additional I-to-T amino acid substitution ure S5C). The 1918-like avian HA induced membrane fusion at
at position 187 of HA (Table 2), which is located at the receptor- pH 5.3 or below, whereas membrane fusion was observed at
binding site of HA (Figure 4A). For the pair 3 contact ferret, the pH 5.6 or below in the cells expressing 1918 or DK/ALB HA
recovered virus possessed an additional T-to-I mutation at posi- (Figure S5C). None of the amino acid changes at positions
tion 232 of NP (Table 2). Taken together, our results demonstrate 89, 113, 190, or 225 affected the fusion pH of the 1918-like
that ten amino acid substitutions (E627K and A684D in PB2; avian HA (i.e., the value at which HA was activated was pH
E89D, S113N, I187T, E190D, G225D, and D265V in HA; V253M 5.3) (Figure S5C), suggesting that the amino acid changes
in PA; and T232I in NP) may be associated with efficient 1918- found in the HA of the 1918-like avian transmissible mutant
like avian virus transmission in ferrets. have no effect on the pH at which fusion of the 1918-like avian
HA is activated.
The Effects of Amino Acid Changes in the HA of the Our previous studies demonstrated that H5 transmissible
Transmissible 1918-like Avian Viruses on mutants exhibit considerable tolerance to high temperature
Receptor-Binding Specificity and HA Stability compared with a nontransmissible H5 mutant, suggesting a
To better understand the molecular mechanisms behind role for HA stabilization in efficient replication and transmission
the enhanced replicative ability and transmissibility of 1918-like in ferrets (de Vries et al., 2014; Imai et al., 2012). Therefore, we
avian virus carrying HA and PB2 with the amino acid substitu- next tested whether the identified amino acid changes in HA
tions found in the transmissible 1918-like avian viruses, we affect its heat stability. For this experiment, we used the same
examined the effects of these amino acid changes on the func- set of recombinant viruses that were used in the glycan array
tions of HA and the viral polymerases. First, we conducted a described earlier. We tested the loss of hemagglutination activity
glycan microarray to examine the receptor specificity of the and infectivity after incubation at 50
C for various times (Figures
1918-like avian virus HA possessing human-type amino acid S5D and S5E). Introduction of the 190D/225D mutation into
substitutions. For this experiment, we used reassortant viruses 1918-like HA resulted in faster loss of hemagglutination activity,
possessing HA segments derived from 1918, DK/ALB, 1918- whereas the additional introduction of the HA-89D or HA-89D/
like avian HA-190D/225D, 1918-like avian HA-89D/190D/225D, 113N mutation reversed this trend (Figure S5D). The HA-190D/
or 1918-like avian HA-89D/113N/190D/225D virus, their NA 225D virus lost its infectivity rapidly after a 180 min incubation
segment from 1918-like avian virus, and their remaining genes at 50
C (6.8 log10 decrease in virus titers; Figure S5E), whereas
from A/Puerto Rico/8/34 (H1N1; PR8); however, the 1918-like the HA-89D/190D/225D and the HA-89D/113N/190D/225D vi-
avian virus HA and NA genes were tested in the background of ruses were more tolerant of high temperature (3.7 and 4.2 log10
1918-like avian virus since these genes could not be rescued decreases in virus titers under the same conditions, respectively;
in the background of PR8. The 1918 HA bound to a2,6-linked sia- Figure S5E). These results suggest that mutations critical for
losides (human-type receptor) (Figure 4B), whereas the 1918-like human-type receptor binding (i.e., HA-190D/225D) reduced HA
avian (Figure 4C) and DK/ALB (Figure S5A) HAs bound to a2,3- stability, which was restored by additional mutations associated
linked sialosides (avian-type receptor). As reported previously with respiratory droplet transmissibility (i.e., HA-89D and HA-
(Chutinimitkul et al., 2010; Liu et al., 2010; Matrosovich et al., 89D/113N). This trend (i.e., loss of HA stability due to mutations
2000; Watanabe et al., 2011; Yang et al., 2010), Gly residues at conferring human virus receptor recognition and recovery of HA
positions 190 and 225 of HA can confer binding to avian-type re- stability by the acquisition of an additional mutation) is consistent
ceptors, whereas Asp residues at these positions confer binding with our previous findings (de Vries et al., 2014; Imai et al., 2012).
to human-type receptors. Indeed, the 1918-like avian mutant However, the HA heat stability of 1918 HA, as evaluated by hem-
HAs possessing Asp at positions 190 and 225 efficiently bound agglutination activity, was similar to that of the HA-190D/225D
to a2,6Gal-sialylated glycans (human-type receptor) (Figure 4D). virus (Figure S5D), indicating complex interplay among recep-
In addition, mutant HA possessing the additional mutations tor binding specificity, HA stability, and virus transmissibility in
found in the virus recovered from the contact ferret infected ferrets.
with transmitted 1918-like avian PB2-627K:HA-89ED/190D/
225D virus (i.e., HA-89D and HA-113N) also bound to the hu- The Effects of Amino Acid Changes in the Viral RNA
man-type receptor (Figures 4E and S5B). The 1918-like avian Polymerase Complex of the Transmissible 1918-like
mutant HAs exhibited remarkably similar glycan-binding speci- Avian Viruses on Polymerase Activity
ficity. For HAs possessing the E-to-D change at position 89 The viral polymerase complex affects viral replicative ability and
(i.e., HA-89D/113N/190D/225D and HA-89D/190D/225D), there pathogenicity (de Vries et al., 2014; Gabriel et al., 2008; Hatta
was weak binding to glycan 51 that was not detected with HA- et al., 2001; Li et al., 2005; Subbarao et al., 1993). To determine

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A 1918 Virus-inoculated Contact

8 8
6 6
Pair 1

4 4 Pair 2

2 2 Pair 3

0 0
1 3 5 7 9 1 3 5 7 9
B 1918-like avian
8 8
6 6
4 4
2 2
0 0
1 3 5 7 9 1 3 5 7 9

C DK/ALB
8 8
6 6
Virus titers (log10 PFU/ml)

4 4
2 2
0 0
1 3 5 7 9 1 3 5 7 9
D 1918-like avian PB2-627K
8 8
6 6
4 4
2 2
0 0
1 3 5 7 9 1 3 5 7 9
E 1918-like avian PB2-627K:HA-89ED/190D/225D
8 8
6 6
4 4
2 2
0 0
1 3 5 7 9 1 3 5 7 9
F 1918-like avian PB2-627K/684D:HA-89ED/113SN/190D/225D/265DV:PA-253M
8 8
6 6
4 4
2 2
0 0
1 3 5 7 9 1 3 5 7 9
Days after virus-inoculation/contact

(legend on next page)

698 Cell Host & Microbe 15, 692–705, June 11, 2014 ª2014 Elsevier Inc.
Cell Host & Microbe
Pandemic Potential of Avian Influenza Virus

Table 2. Amino Acid Changes in Viral Proteins Acquired during the Transmission of 1918-like Avian Viruses in Ferrets
Amino Acid Sequence and Position
HAa PB2 PA NP
Virus 89 113 187 190 225 265 627 684 253 232
1918b E S T D D G K A V T
1918-like Avianb E S I E G D E A V T
1918-like Avian PB2- 627K:HA-89ED/190D/225Dc E/Dd S I D D D K A V T
1918-like Avian PB2-627K/684D:HA-89ED/ E/Dd S/Nd I D D D K D M T
113SN/190D/225D:PA253Me
1918-like Avian PB2-627K/684D:HA89ED/ E/Dd S/Nd I D D D/Vd K D M T
113SN/190D/225D/265DV:PA253Mf
Pair 1 inoculated E/Dd S/Nd I/Td D D D/Vd K D M T
d
contact E/D S T D D V K D M T
Pair 3 inoculated E/Dd S/Nd I D D D/Vd K D M T
d
contact E/D N I D D D K D M I
See also Figures S3 and S4.
a
Amino acid positions of HA are based on H3 HA numbering.
b
The stock of this egg-grown virus was sequenced.
c
The stock of this MDCK-grown virus was sequenced.
d
Indicates a mixed population.
e
Virus isolated from the nasal washes of the contact ferret in this 1918-like avian PB2-627K:HA-89ED/190D/225D virus group was sequenced.
f
Virus isolated from the nasal washes of the contact ferret in this 1918-like avian PB2-627K:HA-89ED/190D/225D virus group was propagated in MDCK
cells, and the resulting virus stock was sequenced.

whether amino acid changes in PA and/or PB2 affect the viral (Table S4). In contrast, the sera reacted robustly with the 1918
polymerase activity of the 1918-like avian virus, we conducted virus and the mutant 1918-like avian viruses that possessed
a luciferase activity-based minireplicon assay in human 293T additional mutations associated with respiratory droplet trans-
cells as described previously (Ozawa et al., 2007). The poly- missibility (i.e., the 1918-like avian PB2-627K:HA-89ED/190D/
merase activity of the 1918 polymerase complex was signifi- 225D and 1918-like avian PB2-627K/684D:HA-89ED/113SN/
cantly greater than that of the 1918-like avian polymerase com- 190D/225D/265DV:PA-253M viruses), although the HI titers
plex at 33
C and 37
C (Figure S5F; p < 0.05). The PB2-627K against the 1918-related viruses were substantially lower than
mutation strongly increased the polymerase activity of the those against the homologous strain (i.e., A/California/04/2009)
1918-like avian polymerase complex (p < 0.05), whereas (Table S4). Given that the amino acids at positions 190 and
neither PB2-684D nor PA-253M appreciably affected the poly- 225 are located close to the antigenic sites of HA, the amino
merase activity (Figure S5F). These findings suggest that only acid substitutions at these positions may alter the antigenicity
the PB2-627K mutation contributes to the enhanced polymer- of the 1918-like avian virus. We next examined the oseltamivir
ase activity. sensitivity of our test viruses. A previous study showed that
1918 virus was highly susceptible to the NA inhibitor oseltamivir
The Effectiveness of Current Vaccines and Antiviral (Tumpey et al., 2002). The IC50 (half maximal inhibitory concen-
Drugs against the 1918-like Avian Virus tration) value of the 1918-like avian virus (<10 nM) was
From a biosafety perspective, it is important to know whether similar to that of the 1918 virus and that of A/Kawasaki/UT-
current control measures (vaccines and antiviral drugs) are effec- K25/2008 (H1N1; an oseltamivir-sensitive control; Table S5). In
tive against the 1918-like avian virus. We therefore examined contrast, the IC50 values of the oseltamivir-resistant controls,
the reactivity of sera from humans vaccinated with the current A/Kawasaki/IMS22B-955/2003 (H3N2) and A/Osaka/180/2009
influenza vaccine, which includes A/California/04/2009 (H1N1), (H1N1), were 34,600 and 1,360 nM, respectively (Table S5).
against the 1918-like avian virus and its mutants. The HI test re- Because all of the viruses tested in this study possess the NA
sults revealed that these sera reacted poorly with 1918-like avian gene from either the 1918 virus or the 1918-like avian virus, these
virus (i.e., to the same extent as the negative control virus A/ viruses should be highly susceptible to oseltamivir, so appro-
mallard/Gurjev/263/82 [H14N5], which possess an HA that is priate control measures would be available to combat the
antigenically distant from that of the current vaccine strains) 1918-like avian virus used in this study.

Figure 3. Respiratory Droplet Transmission in Ferrets

Groups of three ferrets were infected intranasally with 106 pfu of 1918 (A), 1918-like avian (B), DK/ALB (C), 1918-like avian PB2-627K (D), 1918-like avian PB2-
627K:HA-89ED/190D/225D (E), and 1918-like avian PB2-627K/684D:HA-89ED/113N/190D/225D/265DV:PA-253M (F) viruses. After 1 day, a naive ferret (contact
ferret) was placed in a cage adjacent to each infected ferret. Nasal washes were collected from infected ferrets on day 1 after inoculation and from contact ferrets
on day 1 after cohousing and then every other day (for up to 9 days) for virus titration. The lower limit of detection is indicated by the horizontal dashed line. See
also Figure S4 and Table S3.

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700 Cell Host & Microbe 15, 692–705, June 11, 2014 ª2014 Elsevier Inc.
Cell Host & Microbe
Pandemic Potential of Avian Influenza Virus

Global Distribution and Evolution of 1918-like Avian demonstrates the continued circulation of avian influenza viruses
Viruses that possess 1918 virus-like proteins and may acquire 1918
Wild birds harboring a large pool of influenza A virus genomes virus-like properties.
facilitate the global spread of viruses. Here, we experimentally
demonstrated that an avian influenza virus encoding proteins DISCUSSION
with high homology to the 1918 viral proteins and possessing
a limited number of additional mutations exhibited relatively Here, we conducted experiments to assess the risk represented
high pathogenicity in mammals and transmissibility between by avian influenza viral genes encoding proteins that closely
ferrets (Figure 3 and Table 1), suggesting its potential to cause resemble 1918 virus proteins that still circulate in the avian influ-
a pandemic. To assess the risk of the emergence of such enza viral gene pool. We found that a 1918-like avian virus ex-
avian influenza viruses, we examined the prevalence of avian hibited pathogenicity in mice and ferrets higher than that of an
influenza viruses encoding viral proteins that differed from authentic avian virus. Moreover, we demonstrated that acquisi-
the 1918 virus proteins by a few amino acids. We constructed tion of only a few amino acid substitutions can confer respiratory
phylogenetic trees, with a time series of virus isolation (from droplet transmission to 1918-like avian influenza viruses in a
1990 to 2011), of amino acid sequences for each viral protein ferret model, suggesting that the potential exists for a 1918-
by using the 1918 viral protein as the root sequence (Figures like pandemic virus to emerge at any time from the avian virus
5A and S6). To assess the spatial patterns and divergence gene pool.
of the 1918-like avian viruses, we also generated geographic Generally, in experimental infections, avian influenza viruses
maps and histograms to show the distribution of the amino replicate poorly in humans, and vice versa (Beare and Webster,
acid differences from their respective 1918 viral proteins 1991; Hinshaw et al., 1984; Murphy et al., 1984; Webster et al.,
(Figures 5B, 5C, and S6). The geographic patterns of amino 1978), because host restriction constrains interspecies trans-
acid sequence similarity by gene segment showed that avian mission. Recently, however, a newly emerged H7N9 avian influ-
viruses encoding PB2, PB1, NP, M, and NS genes of closest enza virus appeared to break this host barrier and directly infect
similarity to those of the 1918-like avian virus have pre- humans in China, resulting in a total of 238 confirmed cases and
dominantly circulated in North America and Europe, with 57 fatalities as of January 24, 2014 (Schnirring, 2014). We and
mostly sporadic circulation in other parts of the world (Figures others demonstrated that H7N9 viruses isolated from Chinese
5 and S6). patients possess human-adaptive mutations in PB2 and HA,
Further, we examined whether there are any avian influenza vi- which possibly confer efficient replication of avian influenza vi-
ruses possessing human-type amino acids, such as PB2-627K, ruses in the upper respiratory tract of humans, and caused
HA-190D, or HA-225D. We found that 168 of 4,293 avian PB2 limited respiratory droplet transmission in ferrets (Belser et al.,
genes (4%) possess the PB2-627K mutation and that 142 of 2013; Richard et al., 2013; Watanabe et al., 2013; Zhang et al.,
those 168 are from H5N1 viruses that were isolated mainly 2013; Zhu et al., 2013), raising concerns about the pandemic
from wild and domestic birds in Europe, the Middle East, and potential of the H7N9 virus. Similarly, other avian influenza
Africa; the others are H1N1 viruses isolated from poultry in the viruses, including the 1918-like avian viruses described in this
US (Table S7). The HA-190D mutation was found in 9 of 266 study, have the potential to cause a pandemic if they acquire
avian H1 HA sequences (about 3% of avian H1 HA genes), one such human-adaptive mutations. The worst-case scenario is
that also possessed HA-225D (Table S7). Our BLAST search the emergence of a novel avian influenza virus that exhibits
revealed that all of the avian viruses possessing either HA- high pathogenicity in human, like highly pathogenic avian
190D or HA-225D or both probably originated from swine or H5N1 viruses, and efficient transmissibility in humans, like
human viruses that infected an avian species (Table S7). seasonal influenza viruses. To prepare for such a scenario, it is
Taken together, our results reveal the global distribution of important to understand the molecular mechanisms of pathoge-
avian influenza virus genes encoding proteins that differ from nicity and transmissibility of avian influenza viruses. Such infor-
the 1918 virus proteins by only a few amino acids and the exis- mation provides support for pandemic preparedness activities
tence of avian influenza viruses possessing human-type amino (vaccines and antivirals are effective control measures), demon-
acid residues (i.e., PB2 E627K, HA E190D, and HA G225D). strates the value of continued surveillance of avian influenza
Considering the fact that many of these avian influenza viruses viruses, and emphasizes the need for evaluation and integration
were isolated in recent years (i.e., from 1990 to 2011), our study of improved risk assessment measures.

Figure 4. HA Structural Analysis and Glycan Microarray Analysis

(A) Localization of amino acid changes identified in viruses recovered from ferrets in the transmission study. Shown is the 3D structure of the monomer of A/Brevig
Mission/1/18 (H1N1) HA in complex with human receptor analogs (Protein Data Bank [PDB] ID code 2WRG). A close-up view of the globular head is also shown to
the right. Mutations known to increase affinity to human-type receptors are shown in red (E190D and G225D). Mutations that emerged in HA during replication
and/or transmission in ferrets are shown in green (E89D, S113N, I187T, and D265V). The amino acid changes at positions 89 and 113 are located close to an
amino acid at position 110 (103 with H5 numbering) that was previously found to be associated with the transmissibility of an H5 virus (Herfst et al., 2012). Images
were created with MacPyMOL ([Link]
(B–E) The receptor specificities of viruses possessing 1918 HA (B), 1918-like avian HA (C), 1918-like avian HA-190D/225D (D), and 1918-like avian HA-89D/113N/
190D/225D (E) were assessed by using a glycan microarray containing a diverse library of a2,3- and a2,6-linked sialosides (Xu et al., 2013). Viruses, directly
labeled with biotin, were applied at 128 hemagglutination units/ml for 1 hr and, after washing, were incubated with Streptavidin-Alexa Fluor 647 (1 mg/ml) for 1 hr to
detect bound virus. Error bars represent the SD calculated from six replicate spots of each glycan. A complete list of glycans is found in Table S6. See also
Figure S5.

Cell Host & Microbe 15, 692–705, June 11, 2014 ª2014 Elsevier Inc. 701
 Cell Host & Microbe
Pandemic Potential of Avian Influenza Virus

1990
1991
1992
1993
1994
1995
1996
1997
1998
1999
2000
2001
2002
2003
2004
2005
2006
2007
2008
2009
2010
2011
A
A/BREVIG-MISSION/1/1918-PB2
A/MALLARD/NETHERLANDS/30/2006-A/H1N4-2006-XX-XX
A/BLUE-WINGED-TEAL/OHIO/926/2002-A/H3N8-2002-09-07
A/COMMON-TEAL/NETHERLANDS/1/2005-A/H8N4-2005-XX-XX
A/DUCK/SHANTOU/2954/2003-A/H6N8-2003-XX-XX
A/MALLARD/SWEDEN/88/2002-A/H2N1-2002-12-09
A/MALLARD/MINNESOTA/SG-00164/2007-A/H3N8-2007-09-14
A/MALLARD/CALIFORNIA/6471/2008-A/H3N8-2008-11-02
A/MALLARD/CZECH-REPUBLIC/13577-24K/2010-A/H3N8-2010-09-16
A/MALLARD/JIANGXI/6845/2003-A/H6N1-2003-XX-XX
A/MALLARD/SWEDEN/12/2003-A/H2N3-2003-XX-XX
A/MALLARD/SWEDEN/90/2005-A/H11N9-2005-XX-XX
A/NORTHERN-SHOVELER/GEORGIA/1/2010-A/H2N3-2010-XX-XX
A/BLACK-HEADED-GULL/NETHERLANDS/1/2005-A/H6N8-2005-XX-XX
A/MALLARD/NETHERLANDS/13/1999-A-1999-XX-XX
A/DUCK/ALTAI/1285/1991-A/H5N3-1991-08-15
A/MALLARD/SWEDEN/58/2005-A-2005-XX-XX
A/MALLARD/SWEDEN/41/2002-A/H10N6-2002-11-13
A/GULL/MOSCOW/3100/2006-A/H6N2-2006-XX-XX
A/MALLARD/CALIFORNIA/5491/2009-A/H5N2-2009-07-23
A/MALLARD/STRALSUND/55-4/1981-A/H2N3-1981-XX-XX
A/MALLARD/STRALSUND/55-6/1981-A/H2N3-1981-XX-XX
A/BARNACLE-GOOSE/NETHERLANDS/1/2005-A/H6N2-2005-XX-XX
A/CHICKEN/IRAN/SS1/1998-A/H9N2-1998-XX-XX
A/CHICKEN/IRAN/SS4/2009-A/H9N2-2009-01-27
A/CHICKEN/IRAN/SS6/2010-A/H9N2-2010-12-25
A/CHICKEN/IRAN/SS7/2011-A/H9N2-2011-11-17
A/DUCK/HUNAN/689/2006-A/H5N1-2006-12-15
A/MALLARD/ITALY/4223-2/2006-A/H5N2-2006-11-11
A/MALLARD/SWEDEN/30/2005-A/H6N1-2005-XX-XX
A/CHICKEN/QUEENSLAND/1995-A/H7N3-1995-XX-XX
A/CHICKEN/QUEENSLAND/1994-A/H7N3-1994-12-21
A/AQUATIC-BIRD/KOREA/W113/2006-A/H5N2-2006-11-XX
A/DUCK/VICTORIA/1976-A/H7N7-1976-XX-XX
A/MALLARD/SWEDEN/60/2005-A-2005-XX-XX
A/CHICKEN/NETHERLANDS/2586/2003-A/H7N7-2003-XX-XX
A/DUCK/ITALY/194659/2006-A/H3N2-2006-XX-XX
A/DUCK/TAIWAN/0526/72-A/H6N1-1972-XX-XX
A/BLACK-HEADED-GULL/NETHERLANDS/1/2006-A/H4N5-2006-XX-XX
A/AMERICAN-WIGEON/CALIFORNIA/8121/2008-A/H6N1-2008-10-19
A/NORTHERN-PINTAIL/CALIFORNIA/8470/2008-A/H6N1-2008-10-22
A/MALLARD/CALIFORNIA/8399/2008-A/H6N1-2008-10-22
A/GADWALL/CALIFORNIA/8504/2008-A/H6N1-2008-10-25
A/GADWALL/CALIFORNIA/8535/2008-A/H6N1-2008-10-25
A/NORTHERN-PINTAIL/CALIFORNIA/6763/2008-A/H6N1-2008-11-15
A/MALLARD/NETHERLANDS/12/2008-A/H2N3-2008-XX-XX
A/MALLARD/ITALY/36/2002-A/H5N3-2002-XX-XX
A/MALLARD/CZECH-REPUBLIC/15008-11K/2009-A/H5N3-2009-09-09
A/MALLARD/MARYLAND/209/2001-A/H10N7-2001-XX-XX
A/MALLARD/MARYLAND/214/2001-A/H10N7-2001-XX-XX
A/CHICKEN/HEBEI/1/2002-A/H7N2-2002-XX-XX
A/DUCK/SHANTOU/2823/2006-A/H6N2-2006-02-XX
A/BEWICKS-SWAN/NETHERLANDS/2/2005-A/H6N8-2005-XX-XX
A/EURASIAN-WIGEON/NETHERLANDS/3/2005-A/H9N2-2005-XX-XX
A/DUCK/HONG-KONG/D73/1976-A/H6N1-1976-XX-XX
A/AUSTRALIAN-SHELDUCK/WESTERN-AUSTRALIA/1756/1983-A/H15N2-1983-XX-XX
A/AUSTRALIAN-SHELDUCK/WESTERN-AUSTRALIA/1762/1979-A/H15N9-1979-XX-XX
A/EURASIAN-COOT/WESTERN-AUSTRALIA/2727/1979-A/H6N2-1979-XX-XX
A/SPUR-WINGED-GOOSE/NIGERIA/210/2008-A/H5N2-2008-04-03
A/DUCK/HOKKAIDO/8/80-A/H3N8-1980-XX-XX
A/DUCK/JIANG-XI/1286/2005-A/H5N2-2005-XX-XX
A/DUCK/EASTERN-CHINA/52/2002-A/H6N2-2002-10-XX
A/MALLARD/NETHERLANDS/14/2007-A/H2N2-2007-XX-XX
A/CHICKEN/ZHEJIANG/HJ/2007-A/H9N2-2007-XX-XX
A/DUCK/GUANGXI/2736/2006-A/H6N8-2006-07-XX
A/AVIAN/JAPAN/8KI0135/2008-A/H6N5-2008-10-08
A/AVIAN/JAPAN/8KI0150/2008-A/H3N8-2008-10-08
A/AVIAN/JAPAN/8KI0180/2008-A/H3N8-2008-10-08
A/MALLARD/HUADONG/S/2005-A/H5N1-2005-XX-XX
A/DUCK/MEMPHIS/546/1974-A/H11N9-1974-XX-XX
A/MALLARD/OHIO/83/1986-A/H4N6-1986-10-24
A/PEKING-DUCK/WAGUN/1689-4/1985-A/H2N3-1985-XX-XX
A/SOFTBILL/CA/33445-158/1992-A/H7N1-1992-XX-XX
A/DUCK/BEIJING/61/05-A/H3N8-2005-XX-XX
A/MALLARD/CZECH-REPUBLIC/14516/2007-A/H3N8-2007-09-17
A/GOOSE/ZAMBIA/06/2008-A/H3N8-2008-07-XX
A/DUCK/ZAMBIA/12/2009-A/H11N9-2009-09-XX
A/CHICKEN/FRANCE/03426/2003-A/H5N2-2003-XX-XX
A/CHICKEN/BRESCIA/1902-A/H7N7-1902-XX-XX
A/CHICKEN/BRESCIA/1902-A/H7N7-1902-XX-XX
A/DUCK/POTSDAM/1402-6/1986-A/H5N2-1986-XX-XX
A/DUCK/EASTERN-CHINA/47/2008-A/H6N2-2008-04-XX
A/DUCK/EASTERN-CHINA/46/2008-A/H6N2-2008-04-XX
A/DUCK/EASTERN-CHINA/13/2010-A/H6N6-2010-03-XX
A/DUCK/EASTERN-CHINA/16/2010-A/H6N6-2010-04-XX
A/GOOSE/EASTERN-CHINA/17/2010-A/H6N6-2010-04-XX
A/SWAN/SLOVENIA/53/2009-A/H7N7-2009-01-20
A/DUCK/ITALY/551/2000-A/H7N1-2000-XX-XX
A/TURKEY/ITALY/1084/2000-A/H7N1-2000-02-10
A/CHICKEN/ITALY/2335/2000-A/H7N1-2000-03-31
A/OSTRICH/ITALY/2332/00-A/H7N1-2000-XX-XX
A/QUAIL/ITALY/396/2000-A/H7N1-2000-01-19
A/TURKEY/ITALY/2984/2000-A/H7N1-2000-05-08
A/CHICKEN/ITALY/5093/99-A/H7N1-1999-XX-XX
A/CHICKEN/ITALY/5093/99-A/H7N1-1999-XX-XX
A/TURKEY/ITALY/977/99-A/H7N1-1999-XX-XX
A/TURKEY/ITALY/3185/99-A/H7N1-1999-09-03
A/TURKEY/ITALY/4301/1999-A/H7N1-1999-11-22
A/CHICKEN/ITALY/322/2001-A/H7N1-2001-XX-XX
A/MALLARD/ALASKA/708/2005-A/H8N4-2005-08-19
A/MALLARD/HEI-LONGJIANG/131/2006-A/H6N2-2006-XX-XX
A/CHICKEN/GUANGDONG/ZHJ/2011-A/H9N2-2011-06-XX
A/CHICKEN/FUJIAN/G9/2009-A/H9N2-2009-12-XX
A/CHICKEN/SHANDONG/SG2/2009-A/H9N2-2009-11-27
A/CHICKEN/TIBET/S4/2009-A/H9N2-2009-09-10
A/CHICKEN/TIBET/S1/2009-A/H9N2-2009-09-10
A/DUCK/TIBET/S2/2009-A/H9N2-2009-09-10
A/PINTAIL-DUCK/ALB/86/1976-A/H3N2-1976-08-13
A/MALLARD/ALBERTA/77/1977-A/H2N3-1977-08-08
A/MALLARD/ALBERTA/300/1977-A/H4N3-1977-08-30
A/MALLARD-DUCK/ALB/761/1978-A/H6N8-1978-08-25
A/RUDDY-TURNSTONE/DELAWARE-BAY/261/1999-A/H9N7-1999-05-24
A/SHOREBIRD/DELAWARE-BAY/210/1999-A/H3N2-1999-05-24
A/MALLARD/ALBERTA/156/2001-A/H3N8-2001-08-06
A/MALLARD/ALBERTA/160/2001-A/H3N8-2001-08-06
A/REDHEAD/ALBERTA/192/2002-A/H3N6-2002-08-16
A/NORTHERN-PINTAIL/ALBERTA/11701/2005-A/H3N8-2005-08-05
A/GREEN-WINGED-TEAL/NOVA-SCOTIA/14917/2005-A/H6N1-2005-09-14
A/NORTHERN-SHOVELER/INTERIOR-ALASKA/6MP1339/2006-A/H3N8-2006-09-09
A/MALLARD/OREGON/44221-105/2006-A/H3N6-2006-10-14
A/MALLARD/MINNESOTA/SG-00045/2007-A/H4N6-2007-08-03
A/MALLARD/MINNESOTA/SG-00060/2007-A/H4N6-2007-08-03
A/MALLARD/ALBERTA/284/2007-A-2007-08-04
A/NORTHERN-PINTAIL/ALASKA/7MP1393/2007-A/H4N6-2007-08-14
A/MALLARD/ALBERTA/115/2007-A-2007-08-22
A/MALLARD/ALBERTA/108/2007-A-2007-08-22
A/MALLARD-BLACK-DUCK-HYBRID/NOVA-SCOTIA/02330/2007-A/H4N6-2007-08-26
A/REDHEAD/ALBERTA/337/2007-A-2007-08-28
A/BLUE-WINGED-TEAL/TEXAS/SG-00077/2007-A/H4N6-2007-09-15
A/BLUE-WINGED-TEAL/LOUISIANA/SG-00099/2007-A-2007-09-15
A/BLUE-WINGED-TEAL/LOUISIANA/SG-00218/2007-A/H3N8-2007-09-15
A/BLUE-WINGED-TEAL/TEXAS/SG-00159/2007-A/H4N8-2007-09-16
A/MALLARD/MINNESOTA/SG-00214/2007-A/H6N1-2007-09-16
A/MALLARD/INTERIOR-ALASKA/7MP1718/2007-A/H4N5-2007-09-16
A/GREEN-WINGED-TEAL/NEW-BRUNSWICK/02592/2007-A/H3N8-2007-09-20
A/GREEN-WINGED-TEAL/NEW-BRUNSWICK/02588/2007-A/H3N8-2007-09-20
A/GREEN-WINGED-TEAL/NEW-BRUNSWICK/02586/2007-A/H3N8-2007-09-20
A/GREEN-WINGED-TEAL/NEW-BRUNSWICK/02601/2007-A/H4N6-2007-09-21
A/MALLARD/CALIFORNIA/11032/2008-A/N8-2008-12-03
A/MALLARD/CALIFORNIA/5351/2009-A/H1N1-2009-08-04
A/BLUE-WINGED-TEAL/WISCONSIN/2649/2009-A/H6N1-2009-10-17
A/BLUE-WINGED-TEAL/GUATEMALA/CIP049-11/2009-A/H11N2-2009-11-11
A/BLUE-WINGED-TEAL/GUATEMALA/CIP049-09/2010-A/H5N3-2010-01-31
A/MALLARD/MARYLAND/316/2003-A/H2N3-2003-XX-XX
A/MALLARD/MARYLAND/807/2007-A/H3N8-2007-XX-XX
A/TURKEY/MO/21939/1987-A/H1N1-1987-XX-XX
A/WATERFOWL/COLORADO/457952-2/2006-A/H5N2-2006-XX-XX
A/BLUE-WINGED-TEAL/ALB/286/1977-A/H3N6-1977-08-10
A/MALLARD-DUCK/ALB/161/1977-A/H4N6-1977-09-03
A/MALLARD/ALBERTA/54/1991-A/H3N8-1991-08-09
A/MALLARD/CALIFORNIA/19524-001/2005-A/H6N8-2005-08-14
A/AMERICAN-GREEN-WINGED-TEAL/CALIFORNIA/28855/2007-A/H7N3-2007-02-03
A/BLUE-WINGED-TEAL/MINNESOTA/SG-00028/2007-A/H4N6-2007-08-02
A/AMERICAN-WIGEON/CALIFORNIA/HKWF041C/2007-A/H6N1-2007-10-21
A/CANADA-GOOSE/WYOMING/473197-12/2006-A/H5N2-2006-XX-XX
A/GADWALL/ILLINOIS/3860/2009-A/H6N1-2009-11-15
A/MALLARD/MARYLAND/965/2006-A/H4N6-2006-XX-XX
A/MALLARD/WISCONSIN/428/1975-A/H5N1-1975-XX-XX
A/PINTAIL-DUCK/ALB/599/1979-A/H4N2-1979-08-11
A/MALLARD/MINNESOTA/SG-00057/2007-A/H10N7-2007-08-03
A/NORTHERN-PINTAIL/SASKATCHEWAN/22910/2007-A/H3N2-2007-08-10
A/SHOREBIRD/DELAWARE-BAY/214/1999-A/H3N2-1999-05-24
A/RUDDY-TURNSTONE/DELAWARE-BAY/144/1999-A-1999-05-24
A/SHOREBIRD/DELAWARE-BAY/74/1999-A/H3N2-1999-05-24
A/MALLARD-DUCK/ALB/191/1990-A/H6N3-1990-08-01
A/MALLARD-DUCK/ALB/155/1990-A/H6N3-1990-08-04
A/PINTAIL-DUCK/ALB/159/1977-A/H4N6-1977-08-31
A/MALLARD/ALBERTA/162/2007-A/H12N5-2007-08-03
A/GREEN-WINGED-TEAL/CALIFORNIA/1841/2009-A/H7N3-2009-01-18
A/BLUE-WINGED-TEAL/NEW-BRUNSWICK/03757/2009-A/H3N6-2009-09-14
A/MALLARD-DUCK/ALB/250/1978-A/H6N2-1978-08-07
A/MALLARD-DUCK/ALB/290/1978-A/H6N2-1978-08-09
A/RING-NECKED-DUCK/MINNESOTA/SG-00066/2007-A/H3N8-2007-08-03
A/MALLARD/MINNESOTA/SG-00047/2007-A/H3N8-2007-08-03
A/MALLARD/MINNESOTA/SG-00059/2007-A/H3N8-2007-08-03
A/NORTHERN-PINTAIL/CALIFORNIA/3452/2010-A/H1N1-2010-12-04
A/BLUE-WINGED-TEAL/ALBERTA/120/1991-A/H3N8-1991-08-10
A/SHOREBIRD/DELAWARE-BAY/18/2002-A/H1N9-2002-05-20
A/RUDDY-TURNSTONE/DELAWARE/96/2002-A/H2N9-2002-XX-XX
A/BLUE-WINGED-TEAL/WISCONSIN/3061/2009-A-2009-10-17
A/NORTHERN-SHOVELER/ILLINOIS/3767/2009-A-2009-11-14
A/MALLARD/ALBERTA/192/1993-A/H3N8-1993-08-22
A/MALLARD/CALIFORNIA/1297/2010-A/H4N6-2010-07-30
A/DUCK/NJ/7717-70/1995-A/H1N1-1995-XX-XX
A/MALLARD/ILLINOIS/3048/2009-A/H11N2-2009-10-17
A/NORTHERN-SHOVELER/CALIFORNIA/HKWF115/2007-A/H6N1-2007-10-24
A/GREATER-WHITE-FRONTED-GOOSE/CALIFORNIA/44358-077/2007-A/H6N1-2007-10-27
A/MALLARD/WASHINGTON/44338-120/2007-A/H6N2-2007-09-10
A/AMERICAN-WIGEON/CALIFORNIA/HKWF1174/2007-A/H6N1-2007-12-05
A/MALLARD/ALB/5/1995-A/H10N1-1995-08-01
A/MALLARD/ILLINOIS/3974/2009-A/H5N2-2009-10-17
A/NORTHERN-PINTAIL/ILLINOIS/464067-4/2006-A/H5N9-2006-XX-XX
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/8BM3658/2008-A/H4N6-2008-09-24
A/MALLARD/WASHINGTON/44338-039/2007-A/H6N1-2007-09-04
A/AMERICAN-BLACK-DUCK/PRINCE-EDWARD-ISLAND/02708/2007-A/H4N6-2007-08-23
A/COMMON-SCOTER/MARYLAND/297/2005-A/H10N8-2005-12-05
A/MALLARD/MINNESOTA/348/2000-A/H4N6-2000-XX-XX
A/MALLARD/MN/105/2000-A/H5N5-2000-XX-XX
A/MALLARD/SWEDEN/6/2002-A/H2N3-2002-10-25
A/GREEN-WINGED-TEAL/OHIO/86/1986-A/H6N2-1986-10-26
A/LEAST-SANDPIPER/SOUTH-CENTRAL-ALASKA/3/2007-A/H4N8-2007-05-09
A/BLUE-WINGED-TEAL/MINNESOTA/SG-00038/2007-A/H4N6-2007-08-03
A/BLUE-WINGED-TEAL/GUATEMALA/CIP049-01/2008-A/H7N9-2008-02-07
A/MALLARD/CALIFORNIA/6744/2009-A/H6N1-2009-10-25
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00489/2008-A/H4N6-2008-05-18
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00524/2008-A/H4N6-2008-05-22
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00565/2008-A/H4N6-2008-06-04
A/BLUE-WINGED-TEAL/ALBERTA/380/2007-A-2007-08-28
A/BLUE-WINGED-TEAL/ALBERTA/376/2007-A/H3N6-2007-08-28
A/GREEN-WINGED-TEAL/INTERIOR-ALASKA/6MP0909/2006-A/H3N8-2006-08-17
A/MALLARD/OHIO/407/1987-A/H12N5-1987-XX-XX
A/MALLARD/MINNESOTA/SG-00106/2007-A/H6N2-2007-09-16
A/MALLARD/MINNESOTA/SG-00200/2007-A/H3N8-2007-09-22
A/NORTHERN-SHOVELER/CALIFORNIA/44287-088/2007-A/H10N7-2007-01-24
A/MALLARD/ALBERTA/66/2005-A/H4N6-2005-07-27
A/SHOREBIRD/DELAWARE-BAY/77/2001-A/H6N2-2001-05-17
A/SHOREBIRD/DELAWARE-BAY/113/2001-A-2001-05-17
A/MALLARD/ALB/17/1991-A/H9N2-1991-08-09
A/MALLARD/ALB/124/1991-A/H11N2-1991-08-26
A/MALLARD/OHIO/556/1987-A-1987-11-20
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/8BM3669/2008-A/H4N6-2008-09-25
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/8BM3736/2008-A/H12N5-2008-09-27
A/MALLARD/ALBERTA/54/1993-A/H3N9-1993-08-17
A/MALLARD/WISCONSIN/2576/2009-A/H5N1-2009-10-06
A/BLUE-WINGED-TEAL/TEXAS/SG-00083/2007-A/H3N6-2007-09-16
A/PINTAIL/MN/479/99-A/H3N6-1999-XX-XX
A/MALLARD/ALB/206/1996-A/H6N8-1996-XX-XX
A/MALLARD/INTERIOR-ALASKA/6MP0747/2006-A-2006-08-08
A/MALLARD/MD/185/2003-A/H5N2-2003-XX-XX
A/MALLARD/MARYLAND/190/2001-A/H10N7-2001-XX-XX
A/MALLARD/MARYLAND/307/2002-A/H1N1-2002-XX-XX
A/MALLARD/MARYLAND/881/2002-A/H6N2-2002-08-29
A/MALLARD/OHIO/660/2002-A/H4N6-2002-08-19
A/MALLARD/OHIO/657/2002-A/H4N6-2002-08-19
A/BLACK-DUCK/MARYLAND/834/2002-A/H4N8-2002-08-28
A/DUCK/KOREA/U5-2/2007-A/H3N8-2007-05-03
A/AVIAN/NEW-YORK/448534/2006-A/H5N2-2006-XX-XX
A/AVIAN/NEW-YORK/466812/2006-A/H5N2-2006-XX-XX
A/GADWALL/CALIFORNIA/44287-543/2007-A/H10N7-2007-01-28
A/DUCK/NEW-YORK/465976/2006-A/H5N2-2006-XX-XX
A/DUCK/NEW-YORK/504371/2007-A/H5N2-2007-XX-XX
A/DUCK/PENNSYLVANIA/465759/2006-A/H5N2-2006-XX-XX
A/TURKEY/MN/24552/1982-A/H7N3-1982-XX-XX
A/BLUE-WINGED-TEAL/ALB/221/1978-A/H7N2-1978-08-07
A/MALLARD/ILLINOIS/3051/2009-A/H11N3-2009-10-17
A/TURKEY/MINNESOTA/957/80-A/H6N6-1980-09-02
A/AMERICAN-GREEN-WINGED-TEAL/WISCONSIN/2743/2009-A/H1N1-2009-10-17
A/AMERICAN-GREEN-WINGED-TEAL/ILLINOIS/3443/2009-A-2009-10-31
A/BUFFLEHEAD/CALIFORNIA/JN1016/2006-A/H2N9-2006-12-06
A/MALLARD/INTERIOR-ALASKA/6/2007-A/H4N6-2007-09-06
A/MALLARD/INTERIOR-ALASKA/7MP0512/2007-A/H4N6-2007-09-07
A/MALLARD/MINNESOTA/24/2000-A/H5N1-2000-XX-XX
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00522/2008-A/H3N7-2008-05-22
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00553/2008-A/H3N7-2008-05-23
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00529/2008-A/H3N2-2008-05-28
A/MALLARD/CALIFORNIA/3134/2010-A/H1N1-2010-11-24
A/MALLARD/MISSOURI/MO32/2005-A/H11N9-2005-XX-XX
A/NORTHERN-SHOVELER/CALIFORNIA/HKWF1021/2007-A/H3N7-2007-12-02
A/MALLARD/WASHINGTON/44338-052/2007-A/H3N1-2007-09-12
A/BLUE-WINGED-TEAL/MINNESOTA/SG-00452/2008-A/H4N2-2008-08-20
A/DUCK/HUNAN/1590/2007-A/H6N9-2007-07-XX
A/MALLARD/MINNESOTA/417/2000-A/H3N1-2000-XX-XX
A/MALLARD/MONTANA/456423/2006-A/H5N3-2006-XX-XX
A/SHOREBIRD/DELAWARE-BAY/113/2001-A-2001-05-17
A/RUDDY-TURNSTONE/DELAWARE/81/2001-A/H10N7-2001-XX-XX
A/RED-KNOT/DE/1587/2001-A/H10N7-2001-XX-XX
A/RUDDY-TURNSTONE/DELAWARE/2046/2001-A/H5N7-2001-XX-XX
A/RUDDY-TURNSTONE/NJ/1698/2001-A/H5N7-2001-XX-XX
A/RUDDY-TURNSTONE/DELAWARE/1057/2001-A/H10N7-2001-XX-XX
A/RUDDY-TURNSTONE/NJ/1956/2001-A/H10N7-2001-XX-XX
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/7MP1822/2007-A/H3N8-2007-09-05
A/GREEN-WINGED-TEAL/OHIO/80/1993-A/H6N8-1993-XX-XX
A/GREEN-WINGED-TEAL/INTERIOR-ALASKA/6MP0936R1/2006-A/H3N8-2006-08-17
A/PINTAIL/ALASKA/779/2005-A/H6N8-2005-08-20
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/6MP1080R1/2006-A/H3N8-2006-08-20
A/AMERICAN-WIDGEON/ALASKA/7MP1061/2007-A/H3N8-2007-09-10
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/7MP0345BR2/2007-A/H3N8-2007-08-16
A/NORTHERN-SHOVELER/INTERIOR-ALASKA/7MP1670/2007-A/H3N8-2007-09-02
A/MALLARD/INTERIOR-ALASKA/1/2007-A/H3N8-2007-09-06
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/2/2007-A/H4N8-2007-09-07
A/DUCK/INTERIOR-ALASKA/7MP1591R1/2007-A/H3N8-2007-09-01
A/MALLARD/CALIFORNIA/9573/2008-A/H4N6-2008-11-30
A/RED-KNOT/NEW-JERSEY/SG-00478/2008-A/H12N5-2008-05-16
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00500/2008-A/H12N5-2008-05-16
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00495/2008-A/H12N5-2008-05-17
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00493/2008-A/H12N5-2008-05-18
A/RUDDY-TURNSTONE/DELAWARE/SG-00470/2008-A/H4N6-2008-05-21
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00530/2008-A/H12N5-2008-05-22
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00526/2008-A/H10N7-2008-05-23
A/MALLARD/MINNESOTA/SG-00161/2007-A/H3N8-2007-09-16
A/WIDGEON/ALB/256/1982-A/H6N6-1982-08-13
A/BLUE-WINGED-TEAL/ALB/685/1982-A/H6N4-1982-08-20
A/MALLARD/QUEBEC/11247/2006-A/H3N2-2006-08-24
A/GREEN-WINGED-TEAL/MINNESOTA/SG-00222/2007-A/H6N2-2007-09-16
A/MALLARD/MARYLAND/750/2002-A/H4N6-2002-08-27
A/PINTAIL/ALBERTA/202/2000-A/H10N7-2000-08-18
A/MALLARD/INTERIOR-ALASKA/8BM3627R1/2008-A/H4N6-2008-09-16
A/MALLARD/INTERIOR-ALASKA/8BM3596/2008-A/H3N8-2008-09-15
A/AMERICAN-GREEN-WINGED-TEAL/ILLINOIS/3053/2009-A-2009-10-17
A/RUDDY-TURNSTONE/DELAWARE/891/2006-A/H7N3-2006-XX-XX
A/RUDDY-TURNSTONE/NEW-JERSEY/176/2006-A/H7N3-2006-XX-XX
A/RUDDY-TURNSTONE/NEW-JERSEY/549/2006-A/H7N3-2006-XX-XX
A/RUDDY-TURNSTONE/NEW-JERSEY/166/2006-A/H7N3-2006-XX-XX
A/RUDDY-TURNSTONE/NEW-JERSEY/171/2006-A/H7N3-2006-XX-XX
A/RUDDY-TURNSTONE/NEW-JERSEY/565/2006-A/H7N3-2006-XX-XX
A/RUDDY-TURNSTONE/NEW-JERSEY/589/2006-A/H7N3-2006-XX-XX
A/RUDDY-TURNSTONE/DELAWARE/892/2006-A/H7N7-2006-XX-XX
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/6MP0792/2006-A/H2N3-2006-08-10
A/GREEN-WINGED-TEAL/INTERIOR-ALASKA/6MP1077/2006-A/H3N8-2006-08-20
A/MALLARD/MISSISSIPPI/442/2010-A/H1N1-2010-01-19
A/AMERICAN-WIGEON/CALIFORNIA/8352/2008-A/H12N5-2008-10-19
A/PARROT/GUANGDONG/258/2004-A/H5N2-2004-12-29
A/MALLARD/ALBERTA/233/2003-A/H4N6-2003-08-08
A/MALLARD/MINNESOTA/145/1999-A/H4N6-1999-XX-XX
A/NORTHERN-SHOVELER/WASHINGTON/467923/2006-A/H5N2-2006-XX-XX
A/MALLARD/MARYLAND/178/2002-A/H1N1-2002-XX-XX
A/MALLARD/MD/369/2002-A/H1N1-2002-XX-XX
A/MALLARD/MD/382/2002-A/H1N1-2002-XX-XX
A/MALLARD/MD/352/2002-A/H1N1-2002-XX-XX
A/MALLARD/SWEDEN/84/2003-A/H2N3-2003-XX-XX
A/MALLARD/SWEDEN/99/2002-A/H2N3-2002-XX-XX
A/MALLARD/NETHERLANDS/10/1999-A/H1N8-1999-10-11
A/MALLARD/SWEDEN/3/2002-A/H1N2-2002-10-20
A/MALLARD/SWEDEN/65/2002-A/H10N9-2002-11-17
A/MALLARD/ITALY/37/2002-A/H5N3-2002-XX-XX
A/MALLARD/SWEDEN/5/2005-A/H2N3-2005-XX-XX
A/MALLARD/LOUISIANA/476670-4/2007-A/H5N2-2007-XX-XX
A/MALLARD/MONTANA/458329-2/2006-A/H5N3-2006-XX-XX
A/MALLARD/INDIANA/465297-2/2006-A/H5N2-2006-XX-XX
A/SHOREBIRD/DELAWARE-BAY/277/2000-A/H9N7-2000-05-15
A/AMERICAN-COOT/CALIFORNIA/20116-002/2007-A/H7N3-2007-01-18
A/NORTHERN-PINTAIL/NEW-BRUNSWICK/03547/2009-A/H3N8-2009-09-11
A/MALLARD/MARYLAND/691/2005-A/H3N2-2005-XX-XX
A/MALLARDMARYLAND/708/2005-A/H3N2-2005-XX-XX
A/SANDERLING/DELAWARE-BAY/70/1999-A/H3N8-1999-05-23
A/SANDERLING/DELAWARE-BAY/65/1999-A-1999-05-23
A/SHOREBIRD/DELAWARE-BAY/106/1999-A/H3N2-1999-05-24
A/SHOREBIRD/DELAWARE-BAY/31/1996-A/H9N7-1996-05-14
A/MALLARD/CALIFORNIA/430410/2005-A/H5N9-2005-XX-XX
A/RED-KNOT/NEW-JERSEY/SG-00483/2008-A/H12N5-2008-05-20
A/MALLARD/ALBERTA/190/2006-A-2006-08-04
A/WIGEON/OHIO/379/1988-A/H5N2-1988-10-15
A/GREEN-WINGED-TEAL/PRINCE-EDWARD-ISLAND/03846/2009-A-2009-08-19
A/MALLARD/OHIO/684/2002-A/H4N6-2002-XX-XX
A/MALLARD/OHIO/651/2002-A/H3N8-2002-08-19
A/MALLARD/OHIO/649/2002-A/H3N8-2002-08-19
A/DUCK/NY/6874/1978-A/H3N2-1978-XX-XX
A/MALLARD-DUCK/NEW-YORK/6874/1978-A/H3N2-1978-08-01
A/CHICKEN/JILIN/HJ/2003-A/H5N1-2003-XX-XX
A/DUCK/GUANGXI/585/2005-A/H6N5-2005-XX-XX
A/MALLARD/MANITOBA/23912/2007-A/H4N7-2007-08-24
A/MALLARD/NORTH-CAROLINA/6412-009/2004-A/H10N7-2004-XX-XX
A/SHOREBIRD/DELAWARE-BAY/268/1999-A/H3N8-1999-05-24
A/TURKEY/ITALY/214845/2002-A/H7N3-2002-XX-XX
A/TURKEY/ITALY/8000/2002-A/H7N3-2002-XX-XX
A/TURKEY/ITALY/8912/2002-A/H7N3-2002-XX-XX
A/QUAIL/ITALY/3347/2004-A/H7N3-2004-XX-XX
A/TURKEY/ITALY/251/2003-A/H7N3-2003-XX-XX
A/MALLARD/MARYLAND/182/2006-A/H5N2-2006-XX-XX
A/CHICKEN/HUBEI/WI/1997-A/H5N1-1997-XX-XX
A/TREE-SPARROW/HENAN/2/2004-A/H5N1-2004-XX-XX
A/MALLARD/MARYLAND/1127/2005-A/H3N8-2005-06-27
A/MALLARD/OHIO/468159-4/2006-A/H5N2-2006-XX-XX
A/BLUE-WINGED-TEAL/OHIO/1339/2005-A/H4N6-2005-XX-XX
A/MALLARD/OHIO/1506/2006-A/H3N6-2006-XX-XX
A/SHOREBIRD/DELAWARE/189/2007-A/H5N1-2007-05-23
A/GREEN-WINGED-TEAL/DELAWARE/468157-6/2006-A/H5N2-2006-XX-XX
A/RUDDY-TURNSTONE/DELAWARE/103/2007-A/H5N1-2007-05-22
A/MALLARD/MARYLAND/802/2007-A/H5N1-2007-XX-XX
A/GREEN-WINGED-TEAL/NEW-BRUNSWICK/02630/2007-A/H4N6-2007-09-22
A/MALLARD/OHIO/94/1989-A/H4N2-1989-10-19
A/CINNAMON-TEAL/CALIFORNIA/44287-234/2007-A/H11N9-2007-01-27
A/MALLARD/OHIO/37/1986-A/H2N1-1986-XX-XX
A/BLACK-DUCK/OHIO/194/1986-A/H11N1-1986-11-06
A/NORTHERN-PINTAIL/WISCONSIN/2737/2009-A/H3N2-2009-10-17
A/BALD-EAGLE/VIRGINIA/SG-00154/2008-A-2008-XX-XX
A/RUDDY-TURNSTONE/DE/773/1988-A/H9N6-1988-05-17
A/RUDDY-TURNSTONE/DE/2368/1988-A/H4N9-1988-05-31
A/HERRING-GULL/DE/698/1988-A/H2N1-1988-05-16
A/RUDDY-TURNSTONE/DELAWARE/519/1988-A/H2N1-1988-05-15
A/MALLARD-DUCK/ALB/144/1987-A/H3N2-1987-08-13
A/MALLARD/ALBERTA/144/1987-A-1987-08-13
A/MALLARD/OHIO/275/1987-A/H4N2-1987-10-19
A/GREEN-WINGED-TEAL/OHIO/1844/2005-A/H11N9-2005-XX-XX
A/TURKEY/MINNESOTA/833/1980-A/H4N2-1980-XX-XX
A/MALLARD/SWEDEN/57/2002-A/H2N3-2002-XX-XX
A/MALLARD/CZECH-REPUBLIC/15902-17K/2009-A/H6N2-2009-09-30
A/QUAIL/LEBANON/272/2010-A/H9N2-2010-08-XX
A/MALLARD/CALIFORNIA/1335/2010-A/H10-2010-08-03
A/MALLARD-DUCK/ALB/1012/1979-A/H3N4-1979-08-20
A/MALLARD/BRITISH-COLUMBIA/18311/2006-A/H5N2-2006-08-18
A/MALLARD/CALIFORNIA/1353/2010-A/H10N7-2010-08-06
A/MALLARD/WISCONSIN/2653/2009-A/H4N6-2009-10-17
A/MALLARD/PENNSYLVANIA/9001/1984-A/H2N6-1984-09-06
A/REDHEAD-DUCK/ALBERTA/357/1983-A/H11N9-1983-08-07
A/MALLARD/ALBERTA/881/1984-A/H2N3-1984-08-20
A/NORTHERN-SHOVELER/CALIFORNIA/HKWF1005/2007-A/H10N3-2007-11-02
A/RED-CRESTED-POCHARD/MONGOLIA/1915/2006-A/H3N6-2006-09-19
A/NORTHERN-PINTAIL/SWEDEN/1/2003-A/H2N3-2003-09-17
A/CHICKEN/JENA/4836/1983-A/H2N2-1984-XX-XX
A/DUCK/YOKOHAMA/AQ10/2003-A/H5N1-2003-XX-XX
A/MALLARD/NETHERLANDS/15/1999-A/H11N9-1999-11-13
A/SHOVELER/NETHERLANDS/19/1999-A/H11N9-1999-XX-XX
A/CHICKEN/VICTORIA/224/1992-A/H7N3-1992-07-30
A/QUAIL/ITALY/1117/1965-A/H10N8-1965-XX-XX
A/WOOD-DUCK/NEW-YORK/60/1982-A/H6N8-1982-12-01
A/MALLARD/WASHINGTON/44338-015/2007-A/H4N6-2007-08-29
A/MALLARD/WASHINGTON/44338-014/2007-A/H4N6-2007-08-29
A/GREEN-WINGED-TEAL/INTERIOR-ALASKA/6MP0736/2006-A/H3N8-2006-08-07
A/MALLARD/INTERIOR-ALASKA/6MP0972R1/2006-A/H3N8-2006-08-17
A/GULL/MARYLAND/18/1977-A/H2N9-1977-08-01
A/CHUKAR/NEW-YORK/11653-1/2005-A/H7N2-2005-02-02
A/GUINEA-FOWL/NEW-YORK/23164-3-05/2005-A/H7N2-2005-03-04
A/GUINEA-FOWL/NEW-YORK/11646-3/2005-A/H7N2-2005-02-01
A/CHICKEN/NEW-YORK/16330/2005-A/H7N2-2005-02-16
A/CHICKEN/NEW-YORK/88291-8/2005-A/H7N2-2005-07-20
A/CHICKEN/NEW-YORK/88291-6/2005-A/H7N2-2005-07-20
A/CHICKEN/NEW-YORK/10196-6/2005-A/H7N2-2005-01-31
A/CHICKEN/NEW-YORK/22067-8/2005-A/H7N2-2005-06-27
A/DUCK/NEW-YORK/21211-6/2005-A/H7N2-2005-03-01
A/DUCK/NEW-YORK/98616-5/2005-A/H7N2-2005-06-27
A/DUCK/NEW-YORK/143646-5/2005-A/H7N2-2005-XX-XX
A/GUINEA-FOWL/NEW-YORK/143646-3/2005-A/H7N2-2005-XX-XX
A/CHICKEN/NEW-YORK/143646-2/2005-A/H7N2-2005-XX-XX
A/GUINEA-FOWL/NEW-YORK/19495-6/2006-A/H7N2-2006-02-21
A/GUINEA-FOWL/NEW-YORK/22071/2005-A/H7N2-2005-03-03
A/GUINEA-FOWL/NEW-YORK/31621-8/2005-A/H7N2-2005-03-24
A/PINTAIL/ALASKA/211/2005-A/H1N8-2005-08-10
A/MALLARD/INTERIOR-ALASKA/6MP0163AR2/2006-A/H3N8-2006-08-12
A/MALLARD/INTERIOR-ALASKA/6MP0975/2006-A/H3N8-2006-08-18
A/MALLARD/INTERIOR-ALASKA/6MP0935R1/2006-A/H3N8-2006-08-17
A/MALLARD/INTERIOR-ALASKA/6MP1107R1/2006-A/H3N8-2006-09-08
A/NORTHERN-PINTAIL/ALASKA/7MP0344/2007-A/H3N8-2007-08-16
A/MALLARD/OHIO/99/1989-A/H10N7-1989-10-19
A/RED-KNOT/DELAWARE/1329/2000-A/H10N7-2000-XX-XX
A/MALLARD-DUCK/ALB/797/1983-A/H11N3-1983-08-23
A/MALLARD/ALBERTA/27/2001-A/H7N3-2001-07-26
A/AMERICAN-GREEN-WINGED-TEAL/CALIFORNIA/27790/2007-A/H11N9-2007-01-24
A/NORTHERN-SHOVELER/CALIFORNIA/HKWF1128/2007-A/H2N7-2007-12-07
A/MALLARD/ALBERTA/24/01-A/H7N3-2001-XX-XX
A/BLUE-WINGED-TEAL/ALB/136/1990-A/H4N3-1990-08-22
A/MALLARD/ALBERTA/208/2000-A/H10N7-2000-08-19
A/DUCK/HONG-KONG/293/1978-A/H7N2-1978-XX-XX
A/CHICKEN/NY/12273-11/1999-A/H7N3-1999-XX-XX
A/MALLARD/ALBERTA/226/2004-A/H4N6-2004-08-15
A/MALLARD/MISSOURI/350/2009-A/H11N9-2009-12-06
A/NORTHERN-SHOVELER/WASHINGTON/44249-731/2006-A/H10N7-2006-11-30
A/WILD-DUCK/KOREA/HDR02/2005-A/H1N1-2005-01-XX
A/SPOTBILL-DUCK/XUYI/6/2005-A/H11N2-2005-XX-XX
A/BAIKAL-TEAL/HONGZE/14/2005-A/H11N9-2005-XX-XX
A/RUDDY-TURNSTONE/NEW-JERSEY/398/2002-A/H2N9-2002-XX-XX
A/CHICKEN/NY/14677-13/98-A/H6N2-1998-XX-XX
A/MALLARD/MISSISSIPPI/407/2010-A-2010-01-16
A/CHICKEN/PA/13609/1993-A/H5N2-1993-XX-XX
A/WHITE-FRONT-GOOSE/ALASKA/446843/2006-A/H5N2-2006-XX-XX
A/MALLARD/ALBERTA/34/2007-A/H3N8-2007-08-18
A/MALLARD/ALBERTA/28/2007-A/H3N8-2007-08-18
A/MALLARD/INTERIOR-ALASKA/6MP0983R1/2006-A/H3N8-2006-08-18
A/MALLARD/WASHINGTON/456273-3/2006-A/H5N2-2006-XX-XX
A/PINTAIL/ALASKA/53/2005-A/H3N6-2005-08-08
A/TUNDRA-SWAN/ALASKA/462958/2006-A/H5N2-2006-XX-XX
A/NORTHERN-SHOVELER/ALASKA/7MP1708/2007-A/H3N8-2007-09-03
A/BLUE-WINGED-TEAL/ALB/16/1997-A/H2N9-1997-10-24
A/MALLARD/OHIO/249/1998-A/H6N1-1998-10-17
A/GREEN-WINGED-TEAL/OHIO/203/1998-A/H6N2-1998-10-17
A/GREEN-WINGED-TEAL/OHIO/196/1999-A-1999-10-23
A/MALLARD/MN/330/1999-A/H3N1-1999-XX-XX
A/MALLARD/MN/68/99-A/H3N2-1999-XX-XX
A/MALLARD/ALBERTA/202/1996-A/H2N5-1996-XX-XX
A/MALLARD/INTERIOR-ALASKA/7MP1772/2007-A/H1N1-2007-09-17
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00557/2008-A/H11N5-2008-06-04
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00515/2008-A/H1N7-2008-05-28
A/TUNDRA-SWAN/ALASKA/450959/2006-A/H5N2-2006-XX-XX
A/MALLARD/CALIFORNIA/11119/2008-A/H11N9-2008-12-06
A/NORTHERN-SHOVELER/ALASKA/7MP1669B/2007-A/H3N8-2007-09-02
A/COMMON-MURRE/OREGON/20361-001/2007-A/H10N7-2007-07-24
A/GREEN-WINGED-TEAL/LOUISIANA/272GW/1987-A/H10N2-1987-XX-XX
A/MALLARD/ALBERTA/353/1988-A/H2N3-1988-09-03
A/MALLARD-DUCK/ALB/353/1988-A/H2N3-1988-09-03
A/MALLARD/ALB/126/1991-A/H11N9-1991-08-26
A/MALLARD/OHIO/118/1993-A/H1N1-1993-10-15
A/MALLARD/OHIO/324/1988-A/H4N6-1988-10-13
A/MALLARD/OHIO/353/1986-A-1986-10-30
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00561/2008-A/H11N9-2008-05-18
A/SEMI-PALMATED-SANDPIPER/BRAZIL/43/1990-A/H2N1-1990-04-18
A/DUCK/INTERIOR-ALASKA/7MP1570/2007-A/H4N6-2007-09-01
A/MALLARD/INTERIOR-ALASKA/2/2007-A/H4N6-2007-09-10
A/NORTHERN-SHOVELER/ALASKA/7MP1113/2007-A/H4N6-2007-09-11
A/MALLARD/ALASKA/44244-097/2006-A/H3N8-2006-09-01
A/AMERICAN-GREEN-WINGED-TEAL/INTERIOR-ALASKA/4/2007-A/H3N8-2007-09-07
A/AMERICAN-GREEN-WINGED-TEAL/INTERIOR-ALASKA/3/2007-A/H3N8-2007-10-17
A/NORTHERN-SHOVELER/INTERIOR-ALASKA/8BM3576/2008-A/N3-2008-09-13
A/GREEN-WINGED-TEAL/INTERIOR-ALASKA/8BM3706/2008-A-2008-09-26
A/CINNAMON-TEAL/BOLIVIA/4537/2001-A/H7N3-2001-XX-XX
A/ROSY-BILLED-POCHARD/ARGENTINA/CIP051-557/2007-A/H6N2-2007-05-XX
A/BLUE-WINGED-TEAL/ALB/295/1977-A/H7N3-1977-09-01
A/MALLARD/OHIO/1851/2005-A/H11N1-2005-XX-XX
A/WHISKERED-TERN/EGYPT/04/2004-A/H6N2-2004-12-05
A/TUFTED-DUCK/PT/13771/2006-A/H7N3-2006-03-22
A/DUCK/GUIZHOU/5302/2007-A/H6N2-2007-09-XX
A/BLUE-WINGED-TEAL/OHIO/1387/2005-A/H6N2-2005-XX-XX
A/MALLARD/INTERIOR-ALASKA/8BM2714/2008-A/H8N4-2008-08-14
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/8BM3284R1/2008-A/N4-2008-08-20
A/MALLARD/INTERIOR-ALASKA/8BM3584R1/2008-A/H8N4-2008-09-14
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/8BM2621R1/2008-A/H8N4-2008-08-14
A/MALLARD/INTERIOR-ALASKA/9BM1799/2009-A/H12N5-2009-07-01
A/DUCK/GUIZHOU/888/2006-A/H6N5-2006-02-XX
A/MALLARD/WASHINGTON/20010-002/2006-A/H3N8-2006-10-01
A/RUDDY-TURNSTONE/DELAWARE-BAY/142/1998-A/H2N8-1998-05-18
A/RUDDY-TURNSTONE/DELAWARE/105/1998-A/H6N8-1998-05-18
A/NORTHERN-PINTAIL/WISCONSIN/4198/2009-A/H11N9-2009-12-03
A/MALLARD/OHIO/648/2002-A/H6N2-2002-08-19
A/MALLARD/JIANGXI/12147/2005-A/H6N2-2005-XX-XX
A/WILD-DUCK/KOREA/UP122/2007-A/H1N1-2007-03-XX
A/DUCK/VIETNAM/G32/2008-A/H11N9-2008-01-XX
A/MUTE-SWAN/NETHERLANDS/1/2006-A/H10N7-2006-XX-XX
A/MALLARD/KOREA/1242/2010-A/H10N6-2010-12-XX
A/WILD-BIRD/KOREA/A12/2010-A/H10N1-2010-02-XX
A/MALLARD/CZECH-REPUBLIC/15902-18K/2009-A/H11N9-2009-09-30
A/AQUATIC-BIRD/KOREA/W164/2007-A/H5N2-2007-01-XX
A/AQUATIC-BIRD/KOREA/W74/2005-A/H5N2-2005-XX-XX
A/AQUATIC-BIRD/KOREA/W209/2007-A/H5N3-2007-11-XX
A/AQUATIC-BIRD/KOREA/W206/2007-A/H5N2-2007-11-XX
A/AQUATIC-BIRD/KOREA/W125/2006-A/H5N2-2006-11-XX
A/DUCK/VICTORIA/23/1981-A/H1N1-1981-XX-XX
A/MALLARD/ALBERTA/378/1988-A/H3N8-1988-09-14
A/DUCK/JIANGSU/10-D4/2011-A/H11N3-2011-10-XX
A/DUCK/ZHEJIANG/0611-15/2011-A/H1N3-2011-06-XX
A/DUCK/ZHEJIANG/0611-19/2011-A/H1N3-2011-06-XX
A/BAR-HEADED-GOOSE/MONGOLIA/143/2005-A/H12N3-2005-08-06
A/DUCK/HOKKAIDO/VAC-2/04-A/H7N7-2004-XX-XX
A/RED-NECKED-STINT/AUSTRALIA/4500/1980-A/H3N8-1980-12-06
A/RED-NECKED-STINT/WESTERN-AUSTRALIA/4188/1984-A/H4N8-1984-XX-XX
A/MALLARD/PA/454069-9/2006-A/H5N1-2006-XX-XX
A/MALLARD/ALBERTA/330/2007-A/H4N6-2007-08-28
A/MALLARD/CALIFORNIA/10064/2008-A/H1N6-2008-12-21
A/CHICKEN/OSAKA/AQ58/2001-A/H9N2-2001-XX-XX
A/CHICKEN/ROSTOCK/45/1934-A/H7N1-1933-XX-XX
A/CHICKEN/FPV/ROSTOCK/1934-A/H7N1-1934-XX-XX
A/MALLARD/ALBERTA/782/1986-A/H3N8-1986-08-26
A/DUCK/NEW-YORK/492652/2007-A/H5N2-2007-XX-XX
A/AVIAN/NY/31588-3/00-A/H5N2-2000-XX-XX
A/AQUATIC-BIRD/KOREA/W217/2007-A/H5N2-2007-11-XX
A/PINTAIL-DUCK/ALB/303/1977-A/H10N7-1977-09-03
A/TUNDRA-SWAN/ALASKA/442960/2006-A/H5N2-2006-XX-XX
A/RUDDY-TURNSTONE/DELAWARE-BAY/125/1994-A/H3N8-1994-05-22
A/RUDDY-TURNSTONE/DELAWARE-BAY/158/1994-A/H3N8-1994-05-22
A/RUDDY-TURNSTONE/DELAWARE-BAY/78/1994-A-1994-05-22
A/RUDDY-TURNSTONE/DELAWARE-BAY/176/1994-A/H3N8-1994-05-22
A/RED-KNOT/DELAWARE-BAY/218/1994-A/H3N8-1994-05-23
A/RUDDY-TURNSTONE/DELAWARE-BAY/118/1994-A/H3N8-1994-05-22
A/NORTHERN-SHOVELER/TX/56/2000-A/H10N7-2000-XX-XX
A/SHOREBIRD/DELAWARE-BAY/282/1994-A-1994-05-22
A/CHICKEN/NEW-YORK/3112-1/95-A/H7N2-1995-XX-XX
A/CHICKEN/PA/149092-1/02-A/H7N2-2002-XX-XX
A/CHICKEN/NJ/118878-5/01-A/H7N2-2001-XX-XX
A/TURKEY/VA/158512/2002-A/H7N2-2002-XX-XX
A/RUDDY-TURNSTONE/NJ/327/2004-A/H5N7-2004-XX-XX
A/RUDDY-TURNSTONE/NEW-JERSEY/110/2004-A/H10N7-2004-XX-XX
A/MALLARD-DUCK/MINNESOTA/1979-A/H3N1-1979-XX-XX
A/NORTHERN-PINTAIL/SOUTH-DAKOTA/SG-00456/2008-A/H6N1-2008-08-22
A/CANADA-GOOSE/COLORADO/474037-5/2006-A/H5N2-2006-XX-XX
A/MALLARD/NETHERLANDS/1/2002-A/H4N6-2002-01-07
A/BAERS-POCHARD/HUNAN/414/2010
A/PINTAIL/KOREA/1173/09-A/H7N7-2009-12-XX
A/DUCK/HOKKAIDO/49/98-A/H9N2-1998-XX-XX
A/EMU/NEW-SOUTH-WALES/775/1997-A/H7N4-1997-12-08
A/CHICKEN/TAIWAN/1203/03-A/H6N1-2003-XX-XX
A/CHICKEN/TAIWAN/0705/99-A/H6N1-1999-XX-XX
A/CHICKEN/TAIWAN/2838V/00-A/H6N1-2000-XX-XX
A/CHICKEN/TAIWAN/1215/01-A/H6N1-2001-XX-XX
A/CHICKEN/TAIWAN/1205/01-A/H6N1-2001-XX-XX
A/CHICKEN/TAIWAN/0208/02-A/H6N1-2002-XX-XX
A/CHICKEN/TAIWAN/0320/02-A/H6N1-2002-XX-XX
A/MALLARD/ALBERTA/130/2003-A/H1N1-2003-08-01
A/BLUE-WINGED-TEAL/ALB/243/1977-A/H4N6-1977-08-27
A/MALLARD/ALBERTA/114/1997-A/H3N8-1997-08-05
A/MALLARD/WISCONSIN/4196/2009-A-2009-12-03
A/PINTAIL-DUCK/ALBERTA/211/1980-A/H2N3-1980-08-06
A/MALLARD/SWEDEN/45/2002-A/H11N8-2002-11-13
A/MALLARD/QUEBEC/11040/2006-A/H3N2-2006-08-19
A/CHICKEN/NEW-YORK/439236/2006-A/H5N2-2006-XX-XX
A/MALLARD/MARYLAND/1143/2005-A-2005-XX-XX
A/MALLARD/QUEBEC/11182/2006-A/H4N6-2006-08-24
A/BLUE-WINGED-TEAL/OHIO/566/2006-A/H7N9-2006-XX-XX
A/MALLARD-DUCK/ALBERTA/331/1985-A/H3N6-1985-08-18
A/MALLARD-DUCK/ALBERTA/323/1988-A/H2N1-1988-09-03
A/RUDDY-TURNSTONE/NEW-JERSEY/138/1990-A/H3N6-1990-05-26
A/AMERICAN-GREEN-WINGED-TEAL/WISCONSIN/08OS2270/2008-A/H3N8-2008-10-19
A/MALLARD-DUCK/ALB/321/1988-A/H9N2-1988-09-03
A/MALLARD/ALBERTA/35/1992-A/H3N6-1992-08-11
A/MALLARD-DUCK/ALBERTA/7/1987-A/H8N4-1987-08-04
A/MALLARD/OHIO/298/1987-A/H4N6-1987-10-19
A/MALLARD/MN/158/2000-A/H3N9-2000-XX-XX
A/TURKEY/TX/10-49-89/1989-A/H9N2-1989-XX-XX
A/MALLARD/INTERIOR-ALASKA/7MP0747/2007-A/H1N1-2007-08-10
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/7MP0278/2007-A/H1N1-2007-08-17
A/CHUKAR/MN/14591-7/1998-A/H5N2-1998-XX-XX
A/RUDDY-TURNSTONE/NEW-JERSEY/1143/2000-A/H10N7-2000-XX-XX
A/MALLARD/MARYLAND/232/2001-A/H2N3-2001-XX-XX
A/CHICKEN/NEW-YORK/13828-3/1995-A/H2N2-1995-XX-XX
A/CHICKEN/NEW-YORK/4438-3/1995-A/H2N2-1994-10-12
A/MALLARD/ALBERTA/284/2007-A-2007-08-04
A/GUINEA-FOWL/NEW-JERSEY/7290-19/1994-A/H2N2-1994-12-19
A/DUCK/PENNSYLVANIA/9411697/1996-A/H2N3-1996-XX-XX
A/MALLARD/ALBERTA/12/1993-A-1993-08-20
A/RUDDY-TURNSTONE/DELAWARE/1771/2002-A/H5N9-2002-XX-XX
A/SANDERLING/DELAWARE/882/2002-A/H2N1-2002-XX-XX
A/SANDERLING/DELAWARE/654/2002-A/H6N4-2002-XX-XX
A/SHOREBIRD/DELAWARE-BAY/133/2002-A/H9N1-2002-05-21
A/CHICKEN/TX/167280-4/02-A/H5N3-2002-XX-XX
A/AMERICAN-WIGEON/ALBERTA/215/1992-A/H3N8-1992-08-24
A/MALLARD/MN/280/99-A/H3N5-1999-XX-XX
A/RUDDY-TURNSTONE/DELAWARE-BAY/170/1994-A/H3N2-1994-05-22
A/RUDDY-TURNSTONE/DELAWARE-BAY/81/1993-A/H2N1-1993-05-15
A/SHOREBIRD/DELAWARE-BAY/64/1994-A/H3N2-1994-05-22
A/RED-KNOT/DELAWARE-BAY/252/1994-A/H3N2-1994-05-23
A/RUDDY-TURNSTONE/DELAWARE-BAY/129/1994-A/H2N3-1994-05-22
A/MALLARD/OHIO/668/2002-A/H4N6-2002-XX-XX
A/MUSCOVY-DUCK/NEW-YORK/23165-13/2005-A/H7N2-2005-03-04
A/CHICKEN/NEW-YORK/23165-11/2005-A/H7N2-2005-03-04
A/CHICKEN/NEW-YORK/30732-10/2005-A/H7N2-2005-03-23
A/CHICKEN/MEXICO/31381-7/1994-A/H5N2-1994-XX-XX
A/CHICKEN/MEXICO/15407/1997-A/H5N2-1997-XX-XX
A/CHICKEN/CHIAPAS/28159-488/1995-A/H5N2-1995-XX-XX
A/CHICKEN/PUEBLA/14585-622/1994-A/H5N2-1994-XX-XX
A/CHICKEN/PUEBLA/14587-644/1994-A/H5N2-1994-XX-XX
A/CHICKEN/QUERETARO/14588-19/1995-A/H5N2-1995-01-09
A/CHICKEN/HIDALGO/28159-232/1994-A/H5N2-1994-XX-XX
A/CHICKEN/HIDALGO/28159-232/1995-A/H5N2-1995-XX-XX
A/CHICKEN/MEXICO/31381-3/1994-A/H5N2-1994-XX-XX
A/CHICKEN/JALISCO/28159-600/1995-A/H5N2-1995-XX-XX
A/NORTHERN-SHOVELER/CALIFORNIA/HKWF1046C/2007-A/H3N5-2007-12-02
A/NORTHERN-SHOVELER/CALIFORNIA/HKWF1199/2007-A/H3N5-2007-12-05
A/BLUE-WINGED-TEAL/LA/B228/1986-A/H1N1-1986-XX-XX
A/MALLARD/MARYLAND/1983/2005-A/H6N8-2005-XX-XX
A/MALLARD/SOUTH-DAKOTA/SG-00127/2007-A/H3N2-2007-09-19
A/TURKEY/NC/353568/2005-A/H3N2-2005-XX-XX
A/TURKEY/ONTARIO/FAV-10/2011-A/H3N2-2011-07-21
A/MALLARD/OHIO/2039/2009-A-2009-08-03
A/MALLARD/OHIO/2033/2009-A/H4N9-2009-08-03
A/RUDDY-TURNSTONE/NEW-JERSEY/SG-00558/2008-A/H4N6-2008-06-04
A/MALLARD/CALIFORNIA/1492/2010-A/H6N4-2010-08-10
A/MALLARD-DUCK/ALBERTA/438/1985-A/H3N4-1985-08-20
A/RUDDY-TURNSTONE/NEW-JERSEY/914/2005-A/H3N8-2005-XX-XX
A/SHOREBIRD/DELAWARE-BAY/179/2005-A/H3N6-2005-05-19
A/RUDDY-TURNSTONE/NEW-JERSEY/327/2005-A/H3N6-2005-XX-XX
A/SHOREBIRD/DELAWARE-BAY/443/2005-A-2005-05-21
A/SANDERLING/NJ/1042/2005-A/H3N6-2005-XX-XX
A/SHOREBIRD/DELAWARE/249/2006-A/H9N2-2006-05-22
A/SANDERLING/DELAWARE-BAY/449/2006-A/H9N2-2006-05-23
A/RUDDY-TURNSTONE/DELAWARE-BAY/279/2006-A/H7N3-2006-05-22
A/RUDDY-TURNSTONE/DELAWARE-BAY/282/2006-A/H7N3-2006-05-22
A/MALLARD/CALIFORNIA/5502/2009-A/H5N2-2009-07-23
A/MALLARD/CALIFORNIA/5219/2009-A/H5N2-2009-07-27
A/MALLARD/CALIFORNIA/5192/2009-A/H4N2-2009-07-27
A/MALLARD/CALIFORNIA/5359/2009-A/H5N2-2009-08-04
A/DUCK/KOREA/KJ/2003-A/H3N2-2003-12-21
A/CHICKEN/PENNSYLVANIA/1/1983-A/H5N2-1983-XX-XX
A/DUCK/FRANCE/080032/2008-A/H5N2-2008-01-XX
A/DUCK/KOREA/A39/2008-A/H6N2-2008-10-XX
A/CHICKEN/KOREA/S21/04-A/H9N2-2004-XX-XX
A/DOUBLE-CRESTED-CORMORANT/CALIFORNIA/20119-001/2007-A/H3N8-2007-01-22
A/AVIAN/MISSOURI/465593-7/2006-A/H5N2-2006-XX-XX
A/PINTAIL/ALBERTA/293/1977-A/H2N9-1977-09-03
A/NORTHERN-SHOVELER/CALIFORNIA/HKWF1325/2007-A/H8N4-2007-12-09
A/CHICKEN/NEW-YORK/28263/1989-A/H6N3-1989-05-02
A/GULL/MINNESOTA/945/1980-A/H13N6-1980-XX-XX
A/GULL/MARYLAND/704/1977-A/H13N6-1977-06-11
A/SHOREBIRD/DELAWARE/224/2006-A/H13N9-2006-05-22
A/BLACK-HEADED-GULL/SWEDEN/1/2005-A/H13N8-2005-XX-XX
A/HERRING-GULL/MONGOLIA/454/2008-A/H13N8-2008-09-XX
A/MALLARD/KENTUCKY/472048-2/2006-A/H5N2-2006-XX-XX
A/COMMON-SCOTER/MARYLAND/299/2005-A-2005-12-05
A/MALLARD/NETHERLANDS/02/2000-A/H10N7-2000-XX-XX
A/CHICKEN/ITALY/312/1997-A/H5N2-1997-XX-XX
A/POULTRY/ITALY/330/1997-A/H5N2-1997-XX-XX
A/DUCK/FUJIAN/12371/2005-A/H6N2-2005-XX-XX
A/MUSCOVY-DUCK/VICTORIA/9211-18-1400/1992-A/H3N8-1992-XX-XX
A/DUCK/WESTERN-AUSTRALIA/8346/1987-A/H4N6-1987-XX-XX
A/DUCK/KOREA/109/11-A/H7N7-2011-01-XX
A/DUCK/SHANTOU/1930/2001-A/H5N1-2001-XX-XX
A/DUCK/FUJIAN/5426/2005-A/H6N2-2005-XX-XX
A/DUCK/SHANTOU/17058/2006-A/H6N6-2006-09-XX
A/DUCK/SHANTOU/5480/2005-A/H6N2-2005-XX-XX
A/DUCK/SHANTOU/31/2005-A/H6N6-2005-XX-XX
A/DUCK/SHANTOU/7386/2006-A/H6N2-2006-04-XX
A/DUCK/SHANTOU/5104/2004-A/H6N2-2004-XX-XX
A/DUCK/SHANTOU/6104/2006-A/H6N2-2006-03-XX
A/DUCK/SHANTOU/1080/2007-A/H6N2-2007-06-XX
A/DUCK/HUBEI/5/2010-A/H6N6-2010-XX-XX
A/DUCK/HUBEI/2/2010-A/H6N6-2010-05-22
A/DUCK/SHANTOU/1217/2002-A/H6N2-2002-XX-XX
A/DUCK/SHANTOU/1249/2002-A/H6N2-2002-XX-XX
A/DUCK/SHANTOU/1646/2002-A/H6N2-2002-XX-XX
A/DUCK/SHANTOU/1674/2002-A/H6N2-2002-XX-XX
A/DUCK/SHANTOU/2471/2002-A/H6N2-2002-XX-XX
A/DUCK/SHANTOU/2045/2002-A/H6N2-2002-XX-XX
A/WILD-DUCK/SHANTOU/1651/2000-A/H6N2-2000-XX-XX
A/WILD-DUCK/SHANTOU/311/2001-A/H6N9-2001-XX-XX
A/DUCK/SHANTOU/313/2002-A/H6N2-2002-XX-XX
A/DUCK/SHANTOU/5540/2001-A/H6N2-2001-XX-XX
A/DUCK/SHANTOU/10776/2006-A/H6N2-2006-06-XX
A/DUCK/SHANTOU/6847/2004-A/H6N2-2004-XX-XX
A/DUCK/SHANTOU/2444/2001-A/H6N2-2001-XX-XX
A/DUCK/SHANTOU/14167/2006-A/H6N2-2006-07-XX
A/DUCK/FUJIAN/2749/2007-A/H6N6-2007-07-XX
A/DUCK/FUJIAN/3076/2007-A/H6N6-2007-07-XX
A/DUCK/FUJIAN/3264/2007-A/H6N6-2007-08-XX
A/DUCK/FUJIAN/2087/2007-A/H6N6-2007-07-XX
A/DUCK/FUJIAN/3059/2007-A/H6N6-2007-07-XX
A/DUCK/FUJIAN/5744/2007-A/H6N2-2007-10-XX
A/DUCK/FUJIAN/8320/2007-A/H6N6-2007-12-XX
A/DUCK/FUJIAN/4224/2007-A/H6N6-2007-09-XX
A/DUCK/FUJIAN/7119/2007-A/H6N6-2007-11-XX
A/DUCK/FUJIAN/2018/2007-A/H6N6-2007-07-XX
A/DUCK/SHANTOU/10196/2005-A/H6N2-2005-XX-XX
A/DUCK/SHANTOU/4617/2005-A/H6N2-2005-XX-XX
A/DUCK/SHANTOU/13145/2005-A/H6N2-2005-XX-XX
A/DUCK/SHANTOU/3371/2006-A/H6N2-2006-02-XX
A/DUCK/SHANTOU/4371/2001-A/H6N2-2001-XX-XX
A/DUCK/SHANTOU/481/2003-A/H6N2-2003-XX-XX
A/DUCK/SHANTOU/4893/2001-A/H6N6-2001-XX-XX
A/DUCK/SHANTOU/2195/2003-A/H6N2-2003-XX-XX
A/DUCK/SHANTOU/4380/2001-A/H6N2-2001-XX-XX
A/DUCK/SHANTOU/3667/2004-A/H6N2-2004-XX-XX
A/DUCK/FUJIAN/1565/2007-A/H6N6-2007-06-XX
A/DUCK/FUJIAN/3701/2005-A/H6N2-2005-XX-XX
A/DUCK/FUJIAN/9062/2006-A/H6N6-2006-09-XX
A/DUCK/FUJIAN/3354/2006-A/H6N6-2006-04-XX
A/DUCK/FUJIAN/6605/2006-A/H6N6-2006-07-XX
A/DUCK/FUJIAN/5498/2006-A/H6N6-2006-06-XX
A/DUCK/SHANTOU/20313/2005-A/H6N6-2005-XX-XX
A/DUCK/FUJIAN/1651/2006-A/H6N6-2006-02-XX
A/DUCK/FUJIAN/2281/2006-A/H6N6-2006-03-XX
A/DUCK/FUJIAN/3796/2006-A/H6N6-2006-04-XX
A/DUCK/FUJIAN/416/2006-A/H6N2-2006-01-XX
A/DUCK/SHANTOU/2450/2002-A/H6N2-2002-XX-XX
A/DUCK/FUJIAN/3937/2005-A/H6N2-2005-XX-XX
A/TURKEY/MISHMAR-HASHARON/619/02-A/H9N2-2002-XX-XX
A/CHICKEN/TALMEI-ELAZAR/1304/03-A/H9N2-2003-XX-XX
A/CHICKEN/ISRAEL/1953/2004-A/H9N2-2004-XX-XX
A/AVIAN/ISRAEL/584/2005-A/H9N2-2005-XX-XX
A/CHICKEN/ISRAEL/869/2007-A/H9N2-2007-11-04
A/CHICKEN/ISRAEL/1040/2007-A/H9N2-2007-12-23
A/CHICKEN/ISRAEL/702/2008-A/H9N2-2008-04-21
A/MALLARD/ALBERTA/235/1999-A/H4N6-1999-08-11
A/MALLARD/MD/26/2003-A/H1N1-2003-XX-XX
A/PINTAIL/ALBERTA/84/2000-A/H11N9-2000-08-07
A/NORTHERN-SHOVELER/INTERIOR-ALASKA/8BM3684/2008-A-2008-09-26
A/RUDDY-TURNSTONE/NEW-JERSEY/1851/2001-A/H5N8-2001-XX-XX
A/SHOREBIRD/DE/1346/2001-A/H5N7-2001-XX-XX
A/PINTAIL/ALBERTA/269/2001-A/H4N6-2001-08-14
A/MALLARD/MARYLAND/13/2003-A/H1N1-2003-XX-XX
A/RUDDY-TURNSTONE/DELAWARE-BAY/135/1996-A/H7N3-1996-05-15
A/DUCK/GUANGDONG/1/1996-A/H7N3-1996-XX-XX
A/DUCK/INTERIOR-ALASKA/7MP1582/2007-A/H1N1-2007-09-01
A/NORTHERN-PINTAIL/INTERIOR-ALASKA/7MP1702/2007-A-2007-09-03
A/MALLARD/INTERIOR-ALASKA/8BM2102/2008-A/H1N1-2008-08-09
A/MALLARD/POSTDAM/178-4/83-A/H2N2-1983-XX-XX
A/MALLARD/POTSDAM/177-6/1983-A/H2N2-1983-XX-XX
A/MALLARD/POSTDAM/178-4/83-A/H2N2-1983-XX-XX
A/MALLARD/POSTDAM/178-CIP046-QA6RND2/1983-A/H2N2-1983-XX-XX
A/MALLARD/INTERIOR-ALASKA/9BM1856/2009-A/H4N6-2009-07-02
A/MALLARD/INTERIOR-ALASKA/9BM1809/2009-A/H4N6-2009-07-02
A/MALLARD/INTERIOR-ALASKA/9BM1967/2009-A/H4N6-2009-07-06
A/MALLARD/INTERIOR-ALASKA/9BM1968/2009-A/H4N6-2009-07-06
A/MALLARD/INTERIOR-ALASKA/9BM1964/2009-A/H4N6-2009-07-06
A/MALLARD/INTERIOR-ALASKA/9BM2057/2009-A/H4N6-2009-07-08
A/MALLARD/INTERIOR-ALASKA/9BM2170/2009-A/H4N6-2009-07-10
A/MALLARD/INTERIOR-ALASKA/9BM2168/2009-A/H4N6-2009-07-10
A/MALLARD/WASHINGTON/44242-124/2006-A/H3N2-2006-07-23
A/PINTAIL-DUCK/ALB/219/1977-A/H1N1-1977-09-03
A/AMERICAN-WIGEON/NEW-BRUNSWICK/04492/2007-A/H3N8-2007-09-04
A/AMERICAN-WIGEON/NEW-BRUNSWICK/04500/2007-A/H3N8-2007-09-04
A/CHICKEN/PAKISTAN/UDL-04/2007-A/H9N2-2007-03-23
A/CHICKEN/TRIPURA/105131/2008-A/H9N2-2008-05-29
A/AVIAN/SAUDI-ARABIA/910135/2006-A/H9N2-2006-XX-XX
A/QUAIL/AICHI/2/2009-A/H7N6-2009-03-XX
A/QUAIL/AICHI/6/2009-A/H7N6-2009-03-XX
A/BLACK-DUCK/NEW-JERSEY/1580/1978-A/H2N3-1978-02-04
A/DUCK/HOKKAIDO/9/99-A/H9N2-1999-XX-XX
A/MALLARD/ALTAI/1208/2007-A/H3N6-2007-09-XX
A/MALLARD/NEW-YORK/66861/1978-A/H2N3-1979-01-31
A/DUCK/YANGZHOU/013/2008-A/H6N5-2008-12-15
A/DUCK/THAILAND/CU-LM7283C/2010-A/H7N6-2010-11-03
A/DUCK/THAILAND/CU-LM7288T/2010-A/H7N6-2010-11-03
A/BLACK-DUCK/PERTH/699/1978-A/H3N8-1978-01-26
A/GRAY-TEAL/WESTERN-AUSTRALIA/1840/1979-A/H4N4-1979-XX-XX
A/DUCK/NANJING/19/2010-A/H1N3-2010-03-14
A/CHICKEN/JILIN/HA/2003-A/H5N1-2003-XX-XX
A/CHICKEN/JILIN/9/2004-A/H5N1-2004-XX-XX
A/CHICKEN/JILIN/HE/2002-A/H5N1-2002-XX-XX
A/CHICKEN/KOREA/SH0902/2009-A/H9N2-2009-01-XX
A/KOREAN-NATIVE-CHICKEN/KOREA/K040110/2010-A/H9N2-2010-03-22
A/CHICKEN/KOREA/KNUSWR09/2009-A/H9N2-2009-04-XX
A/DUCK/SHANTOU/168/2007-A/H6N8-2007-05-XX
A/MALLARD/NEW-ZEALAND/449-89/2004-A-2004-XX-XX
A/MALLARD/NEW-ZEALAND/1440-372/2005-A/H4N6-2005-XX-XX
A/MALLARD/NEW-ZEALAND/449-75/2004-A/H1N2-2004-XX-XX
A/HERRING-GULL/DELAWARE/471/1986-A/H2N7-1986-XX-XX
A/LAUGHING-GULL/NEW-JERSEY/798/1986-A/H2N7-1986-05-12
A/LAUGHING-GULL/NJ/798/1986-A/H2N7-1986-05-12
A/HERRING-GULL/DELAWARE-BAY/433/1986-A/H2N7-1986-06-01
A/GOOSE/GUANGDONG/3/1997-A/H5N1-1997-XX-XX
A/GOOSE/GUANGDONG/1/96-A/H5N1-1996-XX-XX
A/DUCK/HONG-KONG/365/1978-A/H4N6-1978-XX-XX
A/CHICKEN/YOKOHAMA/AQ134/2002-A/H9N2-2002-XX-XX
A/DUCK/SHANGHAI/35/2002-A/H5N1-2002-XX-XX
A/BLACK-BILLED-MAGPIE/GUANGXI/30/2005-A/H9N2-2005-05-XX
A/DUCK/FUJIAN/FQ2/2007-A/H9N2-2007-04-01
A/CHICKEN/HENAN/L3/2008-A/H9N2-2008-XX-XX
A/CHICKEN/HEBEI/Y2/2009-A/H9N2-2009-03-XX
A/CHICKEN/HEBEI/C4/2008-A/H9N2-2008-02-XX
A/CHICKEN/HEBEI/A/2007-A/H9N2-2007-11-XX
A/DUCK/GUANGXI/35/2001-A/H5N1-2001-XX-XX
A/DUCK/GUANGXI/XA/2001-A/H5N1-2001-XX-XX
A/PARTRIDGE/SHANTOU/478/2002-A/H5N1-2002-XX-XX
A/CHICKEN/HENAN/16/2004-A/H5N1-2004-XX-XX
A/CHICKEN/JIANGSU/CZ1/2002-A/H5N1-2002-XX-XX
A/WILD-DUCK/HUNAN/211/2005-A/H5N1-2005-XX-XX
A/OSTRICH/SUZHOU/097/2003-A/H5N1-2003-04-16
A/DUCK/HONG-KONG/29861-2/2000-A/H5N1-2000-XX-XX
A/CHICKEN/HUNAN/41/2004-A/H5N1-2004-XX-XX
A/BROWN-HEADED-GULL/THAILAND/VSMU-28-SPK/2005-A/H5N1-2005-XX-XX
A/CHICKEN/THAILAND/ICRC-7356/2010-A/H5N1-2010-03-24
A/DUCK/THAILAND/TS01/2006-A/H5N1-2006-XX-XX
A/OPEN-BILL-STORK/THAILAND/VSMU-20-AYA/2004-A/H5N1-2004-XX-XX
A/CHICKEN/KOHN-KAEN/NIAH330/2004-A/H5N1-2004-XX-XX
A/DUCK/ANG-THONG/71-A/H5N1-2004-XX-XX
A/DUCK/HONG-KONG/821/2002-A/H5N1-2002-XX-XX
A/OPEN-BILLED-STORK/BANGKOK/LBD0511F/04-A/H5N1-2004-XX-XX
A/OPENBILL-STORK/THAILAND/VSMU-12-BKK/2003-A/H5N1-2003-12-30
A/CHICKEN/NARATHIWAT/NIAH1703/2004-A/H5N1-2004-XX-XX
A/CHICKEN/SHANTOU/4059/2002-A/H5N1-2002-XX-XX
A/TREE-SPARROW/THAILAND/VSMU-14-KRI/2005-A/H5N1-2005-XX-XX
A/OPEN-BILLED-STORK/NAKHONSAWAN/BBD3009M/05-A/H5N1-2005-XX-XX
A/OPEN-BILLED-STORK/NAKHONSAWAN/BBD1421J/05-A/H5N1-2005-XX-XX
A/OPEN-BILLED-STORK/NAKHONSAWAN/BBA2911M/05-A/H5N1-2005-XX-XX
A/OPEN-BILLED-STORK/NAKHONSAWAN/BBA2111M/05-A/H5N1-2005-XX-XX
A/OPEN-BILLED-STORK/NAKHONSAWAN/BBD1821J/05-A/H5N1-2005-XX-XX
A/OPEN-BILLED-STORK/NAKHONSAWAN/BBD0404F/2004-A/H5N1-2004-XX-XX
A/DUCK/VIET-NAM/19/2005-A/H5N1-2005-XX-XX
A/DUCK/VIETNAM/942A/2004-A/H5N1-2004-XX-XX
A/DUCK/VIETNAM/942B/2004-A/H5N1-2004-XX-XX
A/CHICKEN/VIET-NAM/6/2005-A/H5N1-2005-XX-XX
A/CHICKEN/VIET-NAM/8/2005-A/H5N1-2005-XX-XX
A/GOLDEN-PHEASANT/THAILAND/VSMU-21-SPB/2005-A/H5N1-2005-XX-XX
A/CHICKEN/SUKHOTHAI/NIAH114843/2008-A/H5N1-2008-XX-XX
A/CHICKEN/VIET-NAM/AG-010/2004-A/H5N1-2004-XX-XX
A/CHICKEN/VIET-NAM/DN-045/2004-A/H5N1-2004-XX-XX
A/CHICKEN/VIET-NAM/CT-018/2004-A/H5N1-2004-XX-XX
A/CHICKEN/VIET-NAM/LD-080/2004-A/H5N1-2004-XX-XX
A/CHICKEN/VIETNAM/NCVD12/2005-A/H5N1-2005-XX-XX
A/MUSCOVY-DUCK/VIETNAM/NCVD02/2005-A/H5N1-2005-XX-XX
A/DUCK/VIETNAM/NCVD-17/2007-A/H5N1-2007-XX-XX
A/DUCK/VIETNAM/NCVD-26/2007-A/H5N1-2007-XX-XX
A/DUCK/VIETNAM/NCVD-1/2007-A/H5N1-2007-XX-XX
A/CHICKEN/VIETNAM/NCVD09/2005-A/H5N1-2005-XX-XX
A/DUCK/VIETNAM/NCVD-25/2007-A/H5N1-2007-XX-XX
A/DUCK/VIETNAM/NCVD-6/2007-A/H5N1-2007-XX-XX
A/MUSCOVY-DUCK/VIETNAM/NCVD-11/2007-A/H5N1-2007-XX-XX
A/DUCK/CAMBODIA/072D6/2011-A/H5N1-2011-07-23
A/DUCK/YUNNAN/4072/2003-A/H5N1-2003-XX-XX
A/DUCK/YUNNAN/5169/2003-A/H5N1-2003-XX-XX
A/DUCK/YUNNAN/5948/2003-A/H5N1-2003-XX-XX
A/QUAIL/YOGJAKARTA/UT1075/2004-A/H5N1-2004-05-25
A/CHICKEN/TARUTUNG/BPPVI/2005-A/H5N1-2005-XX-XX
A/DUCK/EAST-JAVA/UT1107/2004-A/H5N1-2004-08-25
A/CHICKEN/BALI/UT2092/2005-A/H5N1-2005-XX-XX
A/CHICKEN/SULAWESI-SELATAN/UT2093/2005-A/H5N1-2005-XX-XX
A/CHICKEN/EAST-JAVA/UT6023/2006-A/H5N1-2006-XX-XX
A/CHICKEN/SOUTH-KALIMANTAN/UT6029/2006-A/H5N1-2006-XX-XX
A/CHICKEN/EAST-JAVA/UT6019/2006-A/H5N1-2006-XX-XX
A/CHICKEN/EAST-JAVA/UT6031/2007-A/H5N1-2007-XX-XX
A/DUCK/HUNAN/69/2004-A/H5N1-2004-XX-XX
A/DUCK/YUNNAN/5310/2006-A/H5N1-2006-XX-XX
A/GOOSE/YUNNAN/5599/2006-A/H5N1-2006-XX-XX
A/GOOSE/YUNNAN/5769/2006-A/H5N1-2006-XX-XX
A/CHICKEN/LAOS/35/2008-A/H5N1-2008-XX-XX
A/CHICKEN/LAOS/17/2008-A/H5N1-2008-XX-XX
A/CHICKEN/OITA/8/2004-A/H5N1-2004-XX-XX
A/MALLARD/HUADONG/Y/2003-A/H5N1-2003-XX-XX
A/DUCK/HUBEI/WQ/2003-A/H5N1-2003-XX-XX
A/CHICKEN/SHANTOU/3744/2003-A/H5N1-2003-XX-XX
A/CHICKEN/XINJIANG/78/2005-A/H5N1-2005-XX-XX
A/CHICKEN/SHANXI/2/2006-A/H5N1-2006-XX-XX
A/CHICKEN/HEBEI/326/2005-A/H5N1-2005-XX-XX
A/MALLARD/HUADONG/HN/2005-A/H5N1-2005-XX-XX
A/CHICKEN/NINGXIA/24/2006-A/H5N1-2006-XX-XX
A/CHICKEN/JIANGSU/18/2008-A/H5N1-2008-XX-XX
A/CHICKEN/HENAN/A-7/2006-A/H5N1-2006-XX-XX
A/DUCK/LAOS/P0117/2007-A/H5N1-2007-XX-XX
A/DUCK/LAOS/A0617/2007-A/H5N1-2007-XX-XX
A/DUCK/LAOS/NCVD-35/2007-A/H5N1-2007-XX-XX
A/DUCK/LAOS/P0106/2007-A/H5N1-2007-XX-XX
A/DUCK/LAOS/P0050/2007-A/H5N1-2007-XX-XX
A/CHICKEN/NINH-BINH/209/2005-A/H5N1-2005-XX-XX
A/DUCK/HAI-PHONG-/208/2006-A/H5N1-2006-XX-XX
A/DUCK/VIETNAM/208/2005-A/H5N1-2005-12-07
A/DUCK/VIETNAM/207/2005-A/H5N1-2005-12-07
A/DUCK/VIETNAM/215/2005-A/H5N1-2005-12-16
A/GOOSE/YUNNAN/4985/2006-A/H5N1-2006-XX-XX
A/HOUSE-CROW/HONG-KONG/1203/2007-A/H5N1-2007-XX-XX
A/HOUSE-CROW/HONG-KONG/719/2007-A/H5N1-2007-XX-XX
A/AVIAN/HONG-KONG/0719/2007-A/H5N1-2007-XX-XX
A/SCALY-BREASTED-MUNIA/HONG-KONG/2572/2007-A/H5N1-2007-XX-XX
A/WHITE-BACKED-MUNIA/HONG-KONG/828/2007-A/H5N1-2007-XX-XX
A/CHICKEN/VIETNAM/NCVD-44/2007-A/H5N1-2007-XX-XX
A/DUCK/YUNNAN/47/2006-A/H5N1-2006-XX-XX
A/MUSCOVY-DUCK/BAC-NINH/07-69/2007-A/H5N1-2007-XX-XX
A/CHICKEN/BAC-GIANG/07-74/2007-A/H5N1-2007-XX-XX
A/DUCK/NAM-DINH/07-88/2007-A/H5N1-2007-XX-XX
A/DUCK/VIETNAM/G12/2008-A/H5N1-2008-01-XX
A/DUCK/LAO/987/2010-A/H5N1-2010-03-XX
A/CHICKEN/INDIA/CA0303/2011-A/H5N1-2011-03-06
A/CHICKEN/INDIA/CA0302/2011-A/H5N1-2011-03-06
A/LITTLE-EGRET/HONG-KONG/8550/2007-A/H5N1-2007-XX-XX
A/GREY-HERON/HONG-KONG/3088/2007-A/H5N1-2007-XX-XX
A/GOOSE/JIANGSU/K0403/2010-A/H5N1-2010-02-10
A/DUCK/LAO/471/2010-A/H5N1-2010-03-XX
A/CHICKEN/KOREA/YAQ173/2008-A/H5N1-2008-04-09
A/CHICKEN/KOREA/USQ284/2008-A/H5N1-2008-04-28
A/WHOOPER-SWAN/AKITA/1/2008-A/H5N1-2008-XX-XX
A/WHOOPER-SWAN/AOMORI/2/2008-A/H5N1-2008-XX-XX
A/WHOOPER-SWAN/HOKKAIDO/1/2008-A/H5N1-2008-XX-XX
A/CHICKEN/PRIMORJE/1/2008-A/H5N1-2008-04-10
A/BAR-HEADED-GOOSE/MONGOLIA/X53/2009-A/H5N1-2009-XX-XX
A/COMMON-BUZZARD/BULGARIA/38WB/2010-A/H5N1-2010-03-01
A/WHOOPER-SWAN/HOKKAIDO/4/2011-A/H5N1-2011-01-XX
A/CHICKEN/KOREA/IC546/2011-A/H5N1-2011-02-XX
A/BEAN-GOOSE/TYVA/10/2009-A/H5N1-2009-06-22
A/DUCK/VIETNAM/LBM139/2012-A/H5N1-2012-03-07
A/DUCK/ZHEJIANG/2242/2011-A/H5N1-2011-02-XX
A/GOOSE/YUNNAN/5141/2006-A/H5N1-2006-XX-XX
A/DUCK/HUNAN/11/2007-A/H5N1-2007-XX-XX
A/CYGNUS-OLOR/ITALY/808/2006-A/H5N1-2006-XX-XX
A/MALLARD/ITALY/835/2006-A/H5N1-2006-XX-XX
A/GOOSE/SUZDALKA/10/05-A/H5N1-2005-XX-XX
A/DUCK/INDIA/TR-NIV4396/2008-A/H5N1-2008-04-03
A/GOOSE/TRIPURA/103596/2008-A/H5N1-2008-04-01
A/CHICKEN/ASSAM/142007/2008-A/H5N1-2008-12-07
A/CHICKEN/ASSAM/140203/2008-A/H5N1-2008-11-20
A/CHICKEN/GAZA/714/2006-A/H5N1-2006-XX-XX
A/CHICKEN/NIGERIA/SO494/2006-A/H5N1-2006-XX-XX
A/TURKEY/EGYPT/2253-2/2006-A/H5N1-2006-XX-XX
A/DUCK/NOVOSIBIRSK/56/2005-A/H5N1-2005-XX-XX
A/FALCON/SAUDI-ARABIA/D1795/2005-A/H5N1-2005-XX-XX
A/CHICKEN/EGYPT/CL6/2007-A/H5N1-2007-03-XX
A/CHICKEN/EGYPT/C3BR212/2007-A/H5N1-2007-03-XX
A/CHICKEN/LIAONING/23/2005-A/H5N1-2005-XX-XX
A/CHICKEN/INDIA/NIV33491/06-A/H5N1-2006-XX-XX
A/CHICKEN/NANDURBAR/7966/2006-A/H5N1-2006-02-XX
A/CHICKEN/NANDURBAR/7979/2006-A/H5N1-2006-02-XX
A/CHICKEN/BURKINA-FASO/131/2006-A/H5N1-2006-05-23
A/CHICKEN/SUDAN/1784-10/2006-A/H5N1-2006-XX-XX
A/CHICKEN/SUDAN/1784-7/2006-A/H5N1-2006-XX-XX
A/TURKEY/IVORY-COAST/4372-3/2006-A/H5N1-2006-XX-XX
A/HOODED-VULTURE/BURKINA-FASO/2/2006-A/H5N1-2006-XX-XX
A/CHICKEN/EGYPT/1/2009-A/H5N1-2009-XX-XX
A/CHICKEN/RAWALAKOT/NARC2441A/2006-A/H5N1-2006-XX-XX
A/CHICKEN/SIHALA/NARC33034/2006-A/H5N1-2006-XX-XX
A/CHICKEN/WEST-BENGAL/170564/2009-A/H5N1-2009-03-09
A/DUCK/TUVA/01/2006-A/H5N1-2006-XX-XX
A/CHICKEN/INDIA/81766/2008-A/H5N1-2008-01-19
A/CHICKEN/INDIA/WB-NIV92456/2009-A/H5N1-2009-03-10
A/COMMON-GOLDENEYE/MONGOLIA/12/2006-A/H5N1-2006-XX-XX
A/MOUNTAIN-HAWK-EAGLE/KUMAMOTO/1/07-A/H5N1-2007-XX-XX
A/DUCK/KOREA/ASAN5/2006-A/H5N1-2006-XX-XX
A/CHICKEN/KUWAIT/KISR5/2007-A/H5N1-2007-XX-XX
A/CHICKEN/KUWAIT/KISR6/2007-A/H5N1-2007-XX-XX
A/DOMESTIC-DUCK/GERMANY/R1400/2007-A/H5N1-2007-XX-XX
A/CHICKEN/KUWAIT/KISR2/2007-A/H5N1-2007-XX-XX
A/CHICKEN/KUWAIT/KISR3/2007-A/H5N1-2007-XX-XX
A/CHICKEN/BHUTAN/248015/2010-A/H5N1-2010-02-19
A/CHICKEN/BHUTAN/248009/2010-A/H5N1-2010-02-20
A/CYGNUS-OLOR/ASTRAKHAN/AST05-2-3/2005-A/H5N1-2005-12-01
A/SWAN/GERMANY/R65/2006-A/H5N1-2006-XX-XX
A/WHOOPER-SWAN/SCOTLAND/1430/2006-A/H5N1-2006-03-31
A/CYGNUS-OLOR/ASTRAKHAN/AST05-2-10/2005-A/H5N1-2005-11-26
A/CYGNUS-OLOR/ASTRAKHAN/AST05-2-2/2005-A/H5N1-2005-11-26
A/CYGNUS-OLOR/CASPIAN-SEA/2006-A/H5N1-2006-XX-XX
A/CYGNUS-OLOR/ASTRAKHAN/AST05-2-7/2005-A/H5N1-2005-XX-XX
A/CHICKEN/NIGERIA/1047-62/2006-A/H5N1-2006-XX-XX
A/GUINEA-FOWL/NIGERIA/957-12/2006-A/H5N1-2006-XX-XX
A/CHICKEN/NIGERIA/FA7/2006-A/H5N1-2006-XX-XX
A/CHICKEN/NIGERIA/1047-54/2006-A/H5N1-2006-XX-XX
A/CHICKEN/NIGERIA/1071-7/2007-A/H5N1-2007-XX-XX
A/CHICKEN/NIGERIA/1071-9/2007-A/H5N1-2007-XX-XX
A/CHICKEN/NIGERIA/OG2/2007-A/H5N1-2007-XX-XX
A/GREAT-CORMORANT/TIBET/12/2006-A/H5N1-2006-XX-XX
A/BAR-HEADED-GOOSE/QINGHAI/1-HVRI/2006-A/H5N1-2006-XX-XX
A/CHICKEN/EGYPT/Q1185/2010-A/H5N1-2010-XX-XX
A/TURKEY/KS/4880/1980-A/H1N1-1980-XX-XX
A/SPOT-BILLED-DUCK/KOREA/534/2008-A/H6N1-2008-09-XX
A/CHICKEN/VIET-NAM/10/2005-A/H5N1-2005-XX-XX
A/CHICKEN/VIET-NAM/17/2005-A/H5N1-2005-XX-XX
A/CHICKEN/HUNAN/1793/2007-A/H5N1-2007-01-23
A/CHICKEN/GUANGXI/55/2005-A/H9N2-2005-XX-XX
A/CHICKEN/CHAKWAL/NARC-46/2003-A/H7N3-2003-12-28
A/CHICKEN/CHAKWAL/NARC-148/2004-A/H7N3-2004-05-12
A/CHICKEN/CHAKWAL/NARC-46/2003-A/H7N3-2003-XX-XX
A/CHICKEN/RAWALPINDI/NARC72/2002-A/H7N3-2002-XX-XX
A/CHICKEN/MURREE/NARC-01/1995-A/H7N3-1995-XX-XX
A/RED-NECKED-STINT/AUSTRALIA/2/2004-A/H4N8-2004-XX-XX
A/RUFOUS-NECKED-STINT/JAPAN/6KS0242/2006-A/H4N8-2006-09-02
A/CHICKEN/HEBEI/B1/2001-A/H9N2-2001-XX-XX
A/CHICKEN/HENAN/5/98-A/H9N2-1998-XX-XX
A/CHICKEN/HUBEI/01-MA01/1999-A/H9N2-1999-XX-XX
A/CHICKEN/SHANDONG/1/2008-A/H9N2-2008-01-XX
A/CHICKEN/GUANGDONG/4/00-A/H9N2-2000-XX-XX
A/DUCK/NANJING/1/97-A/H9N2-1997-XX-XX
A/CHICKEN/SHANDONG/6/96-A/H9N2-1996-XX-XX
A/CHICKEN/GUANGDONG/TS/2004-A/H9N2-2004-03-02
A/CHICKEN/HEILONGJIANG/35/00-A/H9N2-2000-XX-XX
A/PHEASANT/HONG-KONG/NT261/00-A/H6N1-2000-XX-XX
A/CHUKKA/HONG-KONG/FY295/00-A/H6N1-2000-XX-XX
A/QUAIL/HONG-KONG/SF550/00-A/H6N1-2000-XX-XX
A/DUCK/VIET-NAM/1/2005-A/H5N1-2005-XX-XX
A/CHICKEN/CHINA/GUANGXI17/2000-A/H9N2-2000-11-20
A/BIRD/GUANGXI/A1/2006-A/H9N2-2006-XX-XX
A/CHICKEN/PAKISTAN/5/99-A/H9N2-1999-XX-XX
A/PHEASANT/HONG-KONG/SSP44/2002-A/H6N1-2002-XX-XX
1990
1991
1992
1993
1994
1995
1996
1997
1998
1999
2000
2001
2002
2003
2004
2005
2006
2007
2008
2009
2010
2011

B C
PB2 (4957 sequences)
Number of sequenced viruses

700
600
500
400
300
200
100
0
0 10 20 30 40 50 60
Number of amino acid differences from 1918 virus

(legend on next page)

702 Cell Host & Microbe 15, 692–705, June 11, 2014 ª2014 Elsevier Inc.
Cell Host & Microbe
Pandemic Potential of Avian Influenza Virus

EXPERIMENTAL PROCEDURES body reactions were visualized with 3,30 -diaminobenzidine tetrahydrochloride
staining by using the DAKO LSAB2 system (DAKOCytomation). Pathological
Cells and Viruses scores were determined as described in the Supplemental Experimental
Human embryonic kidney 293T cells and MDCK cells were maintained in Procedures.
Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf
serum (FCS) and in minimal essential medium (MEM) containing 5% newborn Serological Tests
calf serum, respectively. HeLa cells were maintained in MEM containing 10% Human serum samples obtained from individuals vaccinated with the A/Cali-
FCS. All cells were maintained at 37
C in 5% CO2. fornia/07/2009 (H1N1) hemagglutinin (HA) split vaccine in Japan were used
All viruses used in this study, except for A/duck/Alberta/35/76 (H1N1; DK/ for serological testing. Hemagglutination inhibition titers were determined as
ALB), were generated by using plasmid-driven reverse genetics as described described in the Supplemental Experimental Procedures. All experiments
in the Supplemental Experimental Procedures and a previous report (Neu- with human sera were approved by the Research Ethics Review Committee
mann et al., 1999). Virus growth in cells and eggs was examined as described of the Institute of Medical Science, the University of Tokyo (approval number
in the Supplemental Experimental Procedures. 21-38-1117), and the Institutional Review Board of the University of Wiscon-
sin-Madison.
Experimental Infection of Mice and Ferrets
Female BALB/c mice (5–6 weeks old) (Jackson laboratory) were used to deter- Neuraminidase Inhibition Assay
mine MLD50 values. Under anesthesia, four mice per group were intranasally The sensitivity of viral NA to oseltamivir carboxylate was evaluated by using
inoculated with viruses. Body weight and survival were monitored daily for an NA enzyme inhibition assay based on the methylumbelliferyl-N-acetylneur-
14 days. aminic acid (MUNANA) method as described in previous studies (Gubareva
Female ferrets (5–8 months old) (Triple F Farms), which were serologically et al., 2001; Kiso et al., 2004) and in the Supplemental Experimental
negative by hemagglutination inhibition (HI) assay for currently circulating hu- Procedures.
man influenza viruses, were used in this study. Under anesthesia, six ferrets
per group were intranasally inoculated with 106 pfu (0.5 ml) of viruses. Three Polykaryon Formation Assay
ferrets per group were euthanized on days 3 and 6 postinfection for virological HeLa cells were transfected with expression plasmids expressing various HAs.
and pathological examinations. The virus titers in various organs were deter- Polykaryon formation was induced by exposing cells to low pH buffer and was
mined by use of plaque assays in MDCK cells. For the transmission study, observed as described in the Supplemental Experimental Procedures.
pairs of ferrets were individually housed in adjacent wireframe cages (Showa
Science) that prevented direct and indirect contact between the animals but Thermostability of HA
allowed spread of influenza virus by respiratory droplets, and virus titers in Viruses (128 hemagglutination units in PBS) were incubated at 50
C for
nasal washes collected from each animal were determined by use of plaque the times indicated. Infectivity and hemagglutination activity were determined
assays in MDCK cells as described elsewhere (Imai et al., 2012) and in the by using plaque assays in MDCK cells and the hemagglutination assay with
Supplemental Experimental Procedures. 0.5% turkey blood cells, respectively.
All experiments with mice and ferrets were performed in accordance with
the University of Wisconsin-Madison’s Regulations for Animal Care and Luciferase-Based Minigenome Assay
Use and approved by the Animal Experiment Committee of the University of A luciferase-based minigenome assay was performed to examine viral poly-
Wisconsin-Madison. merase activity as described previously (Ozawa et al., 2007). Briefly, 293T
cells were transfected with plasmids for the expression of viral proteins PA,
Glycan Arrays PB1, PB2, and NP derived from 1918-like avian virus or its mutants and
Glycan array analysis was performed on a glass slide microarray containing 6 with pPolI-WSN-NA-firefly-luciferase. Plasmid pGL4.74[hRuc/TK] (Promega)
replicates of 58 diverse sialic acid-containing glycans including terminal se- served as an internal control for the dual-luciferase assay. After transfection,
quences and intact N-linked and O-linked glycans found on mammalian and the cells were incubated at 33
C or 37
C for 24 hr, and then luciferase activity
avian glycoproteins and glycolipids (Xu et al., 2013). Virus samples were was measured with the dual-luciferase reporter system (Promega) on a
directly labeled with biotin (Ramya et al., 2013) and then applied to the slide GloMax Microplate Luminometer (Promega) according to the manufacturer’s
array; slide scanning to detect virus bound to glycans was conducted as instructions.
described previously (Watanabe et al., 2013) and in the Supplemental Exper-
imental Procedures. A complete list of glycans present on the array is provided Statistical Analysis
in Table S6. Statistical analyses were performed by using ANOVA in GraphPad Prism
version 5.0 (GraphPad); p values of < 0.05 were considered significant.
Pathological Examination
Excised tissues of animal organs preserved in 10% phosphate-buffered Biosafety and Biosecurity
formalin were processed for paraffin embedding and cut into 3 mm thick sec- All experiments with 1918-related viruses were performed in biosafety level
tions. One section from each tissue sample was stained using a standard 3 (BSL3) agriculture containment laboratories. In vitro experiments were
hematoxylin and eosin (H&E) procedure, whereas another one was processed conducted in Class II biological safety cabinets, and transmission experiments
for immunohistological staining with a rabbit polyclonal antibody for type A were conducted in HEPA-filtered ferret isolators (Imai et al., 2012). The research
influenza nucleoprotein (NP) antigen (prepared in our laboratory) that reacts program, procedures, occupational health plan, documentation, security,
comparably with all of the viruses tested in this study. Specific antigen-anti- and facilities are reviewed annually by the University of Wisconsin-Madison

Figure 5. Global Patterns of PB2 Proteins Derived from Avian Influenza Viruses
(A) Phylogenetic tree of 1,022 randomly selected amino acid sequences of PB2 genes derived from avian influenza viruses. The tree was rooted to the PB2
sequence of the 1918 virus, A/Brevig Mission/1/18 (H1N1) (red arrow). The PB2 from A/blue-winged teal/Ohio/926/2002 (H3N8), which is expressed by the 1918-
like avian virus, is indicated by the blue arrow. The year in which the strains were isolated is indicated by horizontal bars to the right of the tree drawn at the same
vertical position as the position of the strain in the tree. The tree and time series are color coded according to the number of amino acid differences from the 1918
virus defined in the histogram shown in (C).
(B) Geographic map indicating locations where the respective viruses shown in (A) were isolated. The map is color coded according to the number of amino acid
differences defined in the histogram shown in (C).
(C) Histogram showing the distribution of the amino acid differences of all avian influenza PB2 proteins from the 1918 PB2 protein and color scheme for (A) and (B)
(red = 0–5 amino acid substitutions; magenta = 6–10; purple = 11–15). See also Figure S6 and Tables S1 and S7.

Cell Host & Microbe 15, 692–705, June 11, 2014 ª2014 Elsevier Inc. 703
 Cell Host & Microbe
Pandemic Potential of Avian Influenza Virus

Responsible Official and at regular intervals by the CDC and the Animal and pandemic influenza A(H1N1) virus affects receptor binding. J. Virol. 84,
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biosecurity and biosafety is described in the Supplemental Experimental M., Xu, R., Yu, W., Kawaoka, Y., et al. (2014). Hemagglutinin receptor
Procedures. specificity and structural analyses of respiratory droplet-transmissible H5N1
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Supplemental Information includes Supplemental Experimental Procedures, Hinshaw, V.S., Bean, W.J., Webster, R.G., Rehg, J.E., Fiorelli, P., Early, G.,
six figures, and seven tables and can be found with this article online at Geraci, J.R., and St Aubin, D.J. (1984). Are seals frequently infected with avian
[Link] influenza viruses? J. Virol. 51, 863–865.
Imai, M., Watanabe, T., Hatta, M., Das, S.C., Ozawa, M., Shinya, K., Zhong, G.,
AUTHOR CONTRIBUTIONS Hanson, A., Katsura, H., Watanabe, S., et al. (2012). Experimental adaptation
of an influenza H5 HA confers respiratory droplet transmission to a reassortant
T.W., G.N., and Y.K. designed the study; T.W., G.Z., M.H., A.H., R.M., S.F., H5 HA/H1N1 virus in ferrets. Nature 486, 420–428.
Y.T., and S.W. performed the experiments; C.A.R., D.F.B., and D.J.S. con- Johnson, N.P., and Mueller, J. (2002). Updating the accounts: global mortality
ducted phylogenetic analysis; N.N., K.T., and H.H. conducted pathologic ex- of the 1918-1920 ‘‘Spanish’’ influenza pandemic. Bull. Hist. Med. 76, 105–115.
amination; T.W., G.Z., C.A.R., N.N., M.H., R.M., S.W., M.I., G.N., H.H., Jones, K.E., Patel, N.G., Levy, M.A., Storeygard, A., Balk, D., Gittleman, J.L.,
J.C.P., D.J.S., and Y.K. analyzed the data; T.W., C.A.R., N.N., R.M., E.A.M., and Daszak, P. (2008). Global trends in emerging infectious diseases. Nature
S.W., M.I., G.N., H.H., J.C.P., and Y.K. wrote the manuscript; and Y.K. over- 451, 990–993.
saw the project. T.W., G.Z., C.A.R. contributed equally to this work. Kiso, M., Mitamura, K., Sakai-Tagawa, Y., Shiraishi, K., Kawakami, C., Kimura,
K., Hayden, F.G., Sugaya, N., and Kawaoka, Y. (2004). Resistant influenza
ACKNOWLEDGMENTS A viruses in children treated with oseltamivir: descriptive study. Lancet 364,
759–765.
We thank Kelly Moore, Naomi Fujimoto, Izumi Ishikawa, and Yuko Sato for Koçer, Z.A., Krauss, S., Stallknecht, D.E., Rehg, J.E., and Webster, R.G.
technical support. We thank Dr. Susan Watson for editing the manuscript. (2012). The potential of avian H1N1 influenza A viruses to replicate and cause
Several glycans in the glycan array were provided by the Consortium for Func- disease in mammalian models. PLoS ONE 7, e41609.
tional Glycomics ([Link] funded by NIGMS
Li, Z., Chen, H., Jiao, P., Deng, G., Tian, G., Li, Y., Hoffmann, E., Webster, R.G.,
grant GM62116. This work was also supported by National Institute of Allergy
Matsuoka, Y., and Yu, K. (2005). Molecular basis of replication of duck H5N1
and Infectious Diseases Public Health Service research grants, by RO1
influenza viruses in a mammalian mouse model. J. Virol. 79, 12058–12064.
AI080598 and R56 AI099275, by ERATO (Japan Science and Technology
Agency), and by the Strategic Basic Research Programs of Japan Science Liu, Y., Childs, R.A., Matrosovich, T., Wharton, S., Palma, A.S., Chai, W.,
and Technology Agency, Japan. C.A.R. was supported by a University Daniels, R., Gregory, V., Uhlendorff, J., Kiso, M., et al. (2010). Altered receptor
Research Fellowship from the Royal Society. specificity and cell tropism of D222G hemagglutinin mutants isolated from fatal
cases of pandemic A(H1N1) 2009 influenza virus. J. Virol. 84, 12069–12074.
Received: January 30, 2014 Matrosovich, M., Tuzikov, A., Bovin, N., Gambaryan, A., Klimov, A., Castrucci,
Revised: March 25, 2014 M.R., Donatelli, I., and Kawaoka, Y. (2000). Early alterations of the receptor-
Accepted: April 24, 2014 binding properties of H1, H2, and H3 avian influenza virus hemagglutinins after
Published: June 11, 2014 their introduction into mammals. J. Virol. 74, 8502–8512.
Matrosovich, M.N., Gambaryan, A.S., and Klenk, H.-D. (2008). Receptor
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