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The pathogenicity of avian H5N1 influenza viruses to mammals has been evolving since the mid-1980s. Here, we demonstrate that H5N1 influenza viruses, isolated from apparently healthy domestic ducks in mainland China from 1999 through 2002, were becoming progressively more pathogenic for mammals, and we present a hypothesis explaining the mechanism of this evolutionary direction. Twenty-one viruses isolated from apparently healthy ducks in southern China from 1999 through 2002 were confirmed to be H5N1 subtype influenza A viruses. These isolates are antigenically similar to A/Goose/Guangdong/1/96 (H5N1) virus, which was the source of the 1997 Hong Kong “bird flu” hemagglutinin gene, and all are highly pathogenic in chickens. The viruses form four pathotypes on the basis of their replication and lethality in mice. There is a clear temporal pattern in the progressively increasing pathogenicity of these isolates in the mammalian model. Five of six H5N1 isolates tested replicated in inoculated ducks and were shed from trachea or cloaca, but none caused disease signs or death. Phylogenetic analysis of the full genome indicated that most of the viruses are reassortants containing the A/Goose/Guangdong/1/96-like hemagglutinin gene and the other genes from unknown Eurasian avian influenza viruses. This study is a characterization of the H5N1 avian influenza viruses recently circulating in ducks in mainland China. Our findings suggest that immediate action is needed to prevent the transmission of highly pathogenic avian influenza viruses from the apparently healthy ducks into chickens or mammalian hosts.
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Duck? Duck? Goose!

MONKS: [chanting] Pie Iesu domine, dona eis requiem.

[bonk]

Pie Iesu domine,...

[bonk]

...dona eis requiem.

[bonk]

Pie Iesu domine,...

[bonk]

...dona eis requiem.

CROWD: A witch! A witch!

[bonk]

A witch! A witch!

MONKS: [chanting] Pie Iesu domine...

CROWD: A witch! A witch! A witch! A witch! We’ve found a witch! A witch! A witch! A witch! A witch! We’ve got a witch! A witch! A witch! Burn her! Burn her! Burn her! We’ve found a witch! We’ve found a witch! A witch! A witch! A witch!

VILLAGER #1: We have found a witch. May we burn her?

CROWD: Burn her! Burn! Burn her! Burn her!

BEDEVERE: How do you know she is a witch?

VILLAGER #2: She looks like one.

CROWD: Right! Yeah! Yeah!

BEDEVERE: Bring her forward.

WITCH: I’m not a witch. I’m not a witch.

BEDEVERE: Uh, but you are dressed as one.

WITCH: They dressed me up like this.

CROWD: Augh, we didn’t! We didn’t...

WITCH: And this isn’t my nose. It’s a false one.

BEDEVERE: Well?

VILLAGER #1: Well, we did do the nose.

BEDEVERE: The nose?

VILLAGER #1: And the hat, but she is a witch!

VILLAGER #2: Yeah!

CROWD: We burn her! Right! Yeaaah! Yeaah!

BEDEVERE: Did you dress her up like this?

VILLAGER #1: No!

VILLAGER #2 and 3: No. No.

VILLAGER #2: No.

VILLAGER #1: No.

VILLAGERS #2 and #3: No.

VILLAGER #1: Yes.

VILLAGER #2: Yes.

VILLAGER #1: Yes. Yeah, a bit.

VILLAGER #3: A bit.

VILLAGERS #1 and #2: A bit.

VILLAGER #3: A bit.

VILLAGER #1: She has got a wart.

RANDOM: [cough]

BEDEVERE: What makes you think she is a witch?

VILLAGER #3: Well, she turned me into a newt.

BEDEVERE: A newt?

VILLAGER #3: I got better.

VILLAGER #2: Burn her anyway!

VILLAGER #1: Burn!

CROWD: Burn her! Burn! Burn her!...

BEDEVERE: Quiet! Quiet! Quiet! Quiet! There are ways of telling whether she is a witch.

VILLAGER #1: Are there?

VILLAGER #2: Ah?

VILLAGER #1: What are they?

CROWD: Tell us! Tell us!...

BEDEVERE: Tell me. What do you do with witches?

VILLAGER #2: Burn!

VILLAGER #1: Burn!

CROWD: Burn! Burn them up! Burn!...

BEDEVERE: And what do you burn apart from witches?

VILLAGER #1: More witches!

VILLAGER #3: Shh!

VILLAGER #2: Wood!

BEDEVERE: So, why do witches burn?

[pause]

VILLAGER #3: B--... ‘cause they’re made of... wood?

BEDEVERE: Good! Heh heh.

CROWD: Oh, yeah. Oh.

BEDEVERE: So, how do we tell whether she is made of wood?

VILLAGER #1: Build a bridge out of her.

BEDEVERE: Ah, but can you not also make bridges out of stone?

VILLAGER #1: Oh, yeah.

RANDOM: Oh, yeah. True. Uhh...

BEDEVERE: Does wood sink in water?

VILLAGER #1: No. No.

VILLAGER #2: No, it floats! It floats!

VILLAGER #1: Throw her into the pond!

CROWD: The pond! Throw her into the pond!

BEDEVERE: What also floats in water?

VILLAGER #1: Bread!

VILLAGER #2: Apples!

VILLAGER #3: Uh, very small rocks!

VILLAGER #1: Cider!

VILLAGER #2: Uh, gra—gravy!

VILLAGER #1: Cherries!

VILLAGER #2: Mud!

VILLAGER #3: Uh, churches! Churches!

VILLAGER #2: Lead! Lead!

ARTHUR: A duck!

CROWD: Oooh.

BEDEVERE: Exactly. So, logically...

VILLAGER #1: If... she... weighs... the same as a duck,... she’s made of wood.

BEDEVERE: And therefore?

VILLAGER #2: A witch!

VILLAGER #1: A witch!

CROWD: A witch! A witch!...

VILLAGER #4: Here is a duck. Use this duck.

[quack quack quack]

BEDEVERE: Very good. We shall use my largest scales.

CROWD: Ohh! Ohh! Burn the witch! Burn the witch! Burn her! Burn her! Burn her! Burn her! Burn her! Burn her! Burn her! Ahh! Ahh...

BEDEVERE: Right. Remove the supports!

[whop]

[clunk]

[creak]

CROWD: A witch! A witch! A witch!

WITCH: It’s a fair cop.

VILLAGER #3: Burn her!

CROWD: Burn her! Burn her! Burn her! Burn! Burn!