From Flu Wiki 2

Forum: Niger H 5 N 1

15 June 2006

niman – at 12:47

HA sequence from Niger available at GenBank

A/duck/Niger/914/2006(H5N1)

Sequence is easily distinguished from Nigeria H5N1 and has a number of polymorphisms not found in other Qinghai sequences, but readily detected in other H5N1’s from Asia.

Another advertisement for recombination.

ricewiki – at 12:51

So this means…. ? That this is yet another “type” or “strain” of H5N1 floating around? Last I understood, there were at least four different distinguishable strains.

(my apologies, I haven’t been keeping up on the strict science lately)

wetDirt – at 13:01

My take on what it means:

There are two lines of thought on how H5N1 spread from Qinghai: One group says it took the train—that is, (commercial) birds were smuggled across the border of china into tibet, then along the route of the Trans-siberia railway into the edge of europe, then across Turkey/Iraq by tramp steamer to northern Africa, where they were smuggled deeper into Africa. If this is to be true, then the sequences found in Africa would be pure Qinghai—that is, they would not show evidence of mixing with local, non H5N1 low-path influenza. They would be relatively pure and unchanged.

Another school of thought says that the principal vector is by migrating wild birds. If this is true, and because birds migrate often in a burst-pause pattern, that is, fly a while, rest up awhile, fly awhile, etc, they have the chance to mingle with local birds carring local avian influenzas. They swap a few infections, and the strains mingle, and now you have a Qinghqi base with small bits that match up with local, non H5N1 avian influenzas.

So which idea is supported by the new sequences? Migratory birds.

Why is this important? Because you only have enough money to fix the problem once, and you better pick the right problem to fix. If the problem is a smuggling problem, you stick your money in education and in border enforcement, and poultry inspections. If the problem is migratory waterfowl, you put the money in surveillance and make sure chicken ranches have good biosecurity and birds are kept away from migrating waterfowl.

And, it gives you information on the makeup of the strains should you need to know for vaccine purposes.

ricewiki – at 13:10

Ah, ok. That’s a great explanation, wetDirt! Well said.

niman – at 13:11

wetDirt,

You are once again spot on with the analysis. This is definitely the Qinhai strain, but each has it’s own travel schedule and can be readily distinguished. If the H5N1 in Nigeria came via trade, then the H5N1 in adjacent Niger would match. If the arrival was by bird, then there would be several varieties.

This sequence points squarly at the migratory birds.

The constellation of shared polymorphisms is like a fingerprint.

This can be seen in the small branches on the tree of Qinghai sequences in Europe in the top and bottom on the tree (tree split for readability purposes).

Kathy in FL – at 13:25

wetDirt – at 13:01

Oh … my … goodness. Now even I can understand this bit of science. <grin> I even have a college education, but you guys normally leave me in the dust holding my aching head.

Seriously though, while I read most of the “high science” threads I usually only understand a particle of what is being said.

Thank you wetdirt. Putting it into a visual context really helped.

niman – at 13:28

The Niger sequence has 6 polymorphisms that are present in a small subset of the Qinghai strain. The 6 polymorphisms are G253A, A524G, C1198T, G1240A, G1447A, T1603C.

Notice that all changes are transitions (replacing a purine with a purine A=G, or a pyrimadine with a pyrimadine C=T). Its really a binomial system.

These are all transitions and all have a clear H5N1 rap sheet.

These changes are NOT random mutations.

They were all acquired from kissin cousins.

niman – at 13:44

Stamped passport of A524G (also in Nigeria isolate)

DQ447199 A/chicken/Egypt/960N3–004/2006 2006 H5N1

DQ406728 A/chicken/Nigeria/641/2006 2006 H5N1

DQ515984 A/Cygnus olor/Czech Republic/5170/2006 2006 H5N1

DQ435200 A/domestic cat/Iraq/820/2006 2006 H5N1

DQ435201 A/domestic goose/Iraq/812/2006 2006 H5N1

DQ659113 A/duck/Niger/914/2006 2006 H5N1

DQ464377 A/Egypt/2782-NAMRU3/2006 2006 H5N1

DQ435202 A/Iraq/207-NAMRU3/2006 2006 H5N1

DQ458992 A/mallard/Bavaria/1/2006 2006 H5N1

DQ449031 A/mallard/Italy/332/2006 2006 H5N1

AB233319 A/bar-headed goose/Mongolia/1/05 2005 H5N1

DQ320898 A/chicken/Guangxi/604/2005 2005 H5N1

DQ449632 A/chicken/Kurgan/05/2005 2005 H5N1

DQ323672 A/chicken/Kurgan/3/2005 2005 H5N1

DQ389158 A/Cygnus olor/Astrakhan/Ast05–2−1/2005 2005 H5N1

DQ434889 A/Cygnus olor/Astrakhan/Ast05–2−10/2005 2005 H5N1

DQ343502 A/Cygnus olor/Astrakhan/Ast05–2−2/2005 2005 H5N1

DQ358746 A/Cygnus olor/Astrakhan/Ast05–2−3/2005 2005 H5N1

DQ363918 A/Cygnus olor/Astrakhan/Ast05–2−4/2005 2005 H5N1

DQ365004 A/Cygnus olor/Astrakhan/Ast05–2−5/2005 2005 H5N1

DQ364996 A/Cygnus olor/Astrakhan/Ast05–2−6/2005 2005 H5N1

DQ363923 A/Cygnus olor/Astrakhan/Ast05–2−7/2005 2005 H5N1

DQ399540 A/Cygnus olor/Astrakhan/Ast05–2−8/2005 2005 H5N1

DQ399547 A/Cygnus olor/Astrakhan/Ast05–2−9/2005 2005 H5N1

DQ320899 A/duck/Guangxi/793/2005 2005 H5N1

DQ449640 A/duck/Kurgan/08/2005 2005 H5N1

DQ320897 A/quail/Guangxi/575/2005 2005 H5N1

DQ320137 A/swan/Astrakhan/1/2005 2005 H5N1

DQ407519 A/turkey/Turkey/1/2005 2005 H5N1

AB233320 A/whooper swan/Mongolia/3/05 2005 H5N1

AB233321 A/whooper swan/Mongolia/4/05 2005 H5N1

AB233322 A/whooper swan/Mongolia/6/05 2005 H5N1

ISDN140936 A/Whooping swan/Mongolia/244/2005 2005 H5N1

AY651346 A/Ck/Hong Kong/31.2/2002 2002 H5N1

AY651350 A/Ck/Hong Kong/3176.3/2002 2002 H5N1

AY651347 A/Ck/Hong Kong/37.4/2002 2002 H5N1

AY651345 A/Gf/Hong Kong/38/2002 2002 H5N1

AY575880 A/pheasant/Hong Kong/675.14/02 2002 H5N1

AY651348 A/SCk/Hong Kong/YU100/2002 2002 H5N1

M18450 A/Duck/Ireland/113/83 1983 H5N8

wetDirt – at 13:46

OK. My take on what this means:

There are two schools of thought about how changes arise in the H5N1 sequence. One school of thought is that these represent random mutations in a virus notorious for random mutations. When you see a change in the sequence, it happened by Cosmic Rays from Outer Space™, and will probably not be repeated elsewhere. Mutations, however, are not expected to be coordinated, that is, it would be wierd for two mutations a certain distance apart to happen the same way at the same time.

Another school of thought is that, while mutations do happen, the bad ones disappear fast, and the good ones are saved, like halloween candy, to be pulled out later. These little packages show up as a ‘chunk’, with specific changes, that remain stable with time, some of which stay geographically isolated, and some of which are widespread.

So what Niman has noticed is that all these changes have a certain pattern: The A purine is swapped for a G purine, or a C pyrimadine with a T pyramidine. Say the purines are bumps and the pyramidines are valleys. Now if you imagine the key-lock thing, what this means is that the little teeth on the key change shape only a tiny bit, because you are swapping one hill for another hill, and a valley for another valley. Similar shapes. And he is noting that it is a consistent pattern of transitions, it’s like the virus saying, hey, look, guys, cool, I can pick the lock with either version of this key, as long as I keep the same general shape.

And he is saying these are no longer random changes, that he sees this pattern over and over and it fits into a larger pattern.

niman – at 13:52

C1198T shared by H5N1 but none are Qinghai sequences

ISDN138756 A/chicken/Malaysia/935/2006 2006 H5N1

DQ659113 A/duck/Niger/914/2006 2006 H5N1

DQ095624 A/Chicken/Yunnan/447/05 2005 H5N1

DQ095625 A/Chicken/Yunnan/493/05 2005 H5N1

AY651322 A/Dk/Indonesia/MS/2004 2004 H5N1

DQ017300 A/duck/Nakornsawan-2–02/2004 2004 H5N1

AY651323 A/Ck/Indonesia/2A/2003 2003 H5N1

AF509026 A/Chicken/Hong Kong/822.1/01 2001 H5N1

AF509028 A/Chicken/Hong Kong/830.2/01 2001 H5N1

AF509029 A/Chicken/Hong Kong/858.3/01 2001 H5N1

AF509030 A/Chicken/Hong Kong/867.1/01 2001 H5N1

AF509033 A/Chicken/Hong Kong/876.1/01 2001 H5N1

AF509031 A/Chicken/Hong Kong/879.1/01 2001 H5N1

AF509034 A/Chicken/Hong Kong/891.1/01 2001 H5N1

AF509035 A/Chicken/Hong Kong/893.2/01 2001 H5N1

AF509024 A/Chicken/Hong Kong/SF219/01 2001 H5N1

AF509017 A/Chicken/Hong Kong/YU562/01 2001 H5N1

AY221529 A/Chicken/Hong Kong/YU562/01 2001 H5N1

AF509018 A/Chicken/Hong Kong/YU563/01 2001 H5N1

AY221528 A/Chicken/Hong Kong/YU822.2/01 2001 H5N1

AY221527 A/Chicken/Hong Kong/YU822.2/01-MB 2001 H5N1

AF509023 A/Pigeon/Hong Kong/SF215/01 2001 H5N1

AF509022 A/Quail/Hong Kong/SF203/01 2001 H5N1

AF509021 A/Silky Chicken/Hong Kong/SF189/01 2001 H5N1

lugon – at 13:57

it would be wierd for two mutations a certain distance apart to happen the same way at the same time

I’m an ignorant looking for enlightment on this one.

Could it be that there are many many many mutations inside the cell, and only those that work get selected, so that it’s not that difficult to see the same wheel reinvented all over the place? I mean, maybe random mutation mechanisms produce a million varieties, and only a hundred are able to make a viable virion, so that most mutations are hidden from view (they never make it on their way out of the cell). With enough random mutations, you might have “twins” that have emerged at random from different branches of the tree: they look like they are relatives, but they are really reinventions of the same thing without family relationship.

I really don’t know.

Thanks.

wetDirt – at 14:01

“niman – at 13:28 The Niger sequence has 6 polymorphisms that are present in a small subset of the Qinghai strain. The 6 polymorphisms are G253A, A524G, C1198T, G1240A, G1447A, T1603C. “

So Henry, Taking any of these polymorphisms at random, how stable are they over time? Are there, say, 100 base pairs that have remained identical for over 5 years? I would like to calculate the probability of that happening using everyone’s quoted mutation rate. I need a nice even number of base pairs because I’m that lazy.

niman – at 14:08

G1240A matches only one Qinghai sequence, but matches low pat from Europe and North America

DQ659113 A/duck/Niger/914/2006 2006 H5N1

AB233319 A/bar-headed goose/Mongolia/1/05 2005 H5N1

DQ309440 A/duck/BritishColumbia/CN26−6/05 2005 H5N2

DQ095629 A/Duck/Fujian/1734/05 2005 H5N1

DQ320897 A/quail/Guangxi/575/2005 2005 H5N1

DQ320893 A/chicken/Guangxi/2439/2004 2004 H5N1

DQ320883 A/duck/Guangxi/1311/2004 2004 H5N1

DQ320885 A/duck/Guangxi/1586/2004 2004 H5N1

DQ320886 A/duck/Guangxi/1681/2004 2004 H5N1

DQ320887 A/duck/Guangxi/1793/2004 2004 H5N1

DQ320890 A/duck/Guangxi/2291/2004 2004 H5N1

DQ320892 A/duck/Guangxi/2396/2004 2004 H5N1

DQ320881 A/goose/Guangxi/1097/2004 2004 H5N1

DQ320882 A/goose/Guangxi/1198/2004 2004 H5N1

DQ320888 A/goose/Guangxi/1832/2004 2004 H5N1

DQ320889 A/goose/Guangxi/2112/2004 2004 H5N1

DQ320891 A/goose/Guangxi/2383/2004 2004 H5N1

AY590577 A/openbill/Thailand/CU-2/2004 2004 H5N1

CY005927 A/shorebird/Delaware/101/2004 2004 H5N7

CY005969 A/shorebird/Delaware/75/2004 2004 H5N7

AY651336 A/Viet Nam/3062/2004 2004 H5N1

AY573917 A/chicken/Taiwan/1209/03 2003 H5N2

AY747609 A/swine/Fujian/1/2003 2003 H5N1

AY296079 A/duck/ME/151895–7A/02 2002 H5N2

AY296080 A/duck/NY/185502/02 2002 H5N2

AY296081 A/duck/NY/186875/02 2002 H5N2

AY296086 A/duck/NY/191255–59/02 2002 H5N8

AY296082 A/duck/NY/191255–79/02 2002 H5N2

AY995886 A/mallard/Sweden/22/02 2002 H5N9

AY995887 A/mallard/Sweden/27/02 2002 H5N9

AY995888 A/mallard/Sweden/28/02 2002 H5N9

AY995889 A/mallard/Sweden/31/02 2002 H5N2

AY995890 A/mallard/Sweden/37/02 2002 H5N9

AY995892 A/mallard/Sweden/40/02 2002 H5N6

AY995893 A/mallard/Sweden/49/02 2002 H5N9

AY995894 A/mallard/Sweden/52/02 2002 H5N9

AY995895 A/mallard/Sweden/58/02 2002 H5N3

AY995896 A/mallard/Sweden/64/02 2002 H5N2

AY995897 A/mallard/Sweden/79/02 2002 H5N9

AY995898 A/mallard/Sweden/80/02 2002 H5N9

AY296083 A/turkey/CA/D0208651-C/02 2002 H5N2

AY075033 A/Duck/Hong Kong/380.5/2001 2001 H5N1

AY296077 A/duck/NJ/117228–7/01 2001 H5N2

AY747617 A/swine/Fujian/F1/2001 2001 H5N1

AY296076 A/unknown/NY/101250–18/01 2001 H5N2

AY296075 A/unknown/NY/9899–6/01 2001 H5N2

AY296073 A/chukkar/NY/51375/00 2000 H5N2

AY296072 A/duck/NY/44018–2/00 2000 H5N2

AY585377 A/duck/Zhejiang/52/2000 2000 H5N1

AJ305306 A/chicken/Italy/8/98 1998 H5N2

AF194169 A/Chicken/Italy/312/97 1997 H5N2

AF194990 A/Chicken/Italy/367/97 1997 H5N2

AF194991 A/Guinea Fowl/Italy/330/97 1997 H5N2

U79448 A/Chicken/Queretaro/7653–20/951659 1995 H5N2

CY006040 A/chicken/Hidalgo/28159–232/1994 1994 H5N2

U28920 A/Emu/Texas/39442/93 (HP progeny) 1993 H5N2

U28919 A/Emu/Texas/39442/93 (non-HP parent) 1993 H5N2

U67783 A/Mallard/Ohio/556/1987 1987 H5N9

U69277 A/Mallard/Wisconsin/944/82 1982 H5

AF082040 A/Duck/Minnesota/1525/81 1981 H5N1

U79454 A/Turkey/Minnesota/3689–1551/81 1981 H5N2

CY005918 A/mallard duck/Alberta/57/1976 1976 H5N2

U79452 A/Mallard/Wisconsin/169/75 1975 H5N3

U79456 A/Turkey/Wisconsin/68 1968 H5N9

AB241613 A/turkey/Ontario/7732/66 1966 H5

M30122 A/turkey/Ontario/7732/66 1966 H5N9

U20460 A/tern/South Africa/61 1961 H5N3

niman – at 14:12

wetDirt,

There is not much of a database on the Qinghai strain (it was just identifed a year ago) but for this HA sequence there are over 1600 bases and only 6 are not common (and all 6 can be easily found in other H5N1s).

wetDirt – at 14:12

Are we talking about an exact match, that is , 100%? If so, then your last entry, a tern from 1961, says that the this polymorphism has remined unchanged for, umm, let’s see, 45 years, hanging around in low-path land.

niman – at 14:28

Some polymorphisms go back to 1918, but that is just a match at that position. The earlier eact matches I talked about are long stretches that including a long string of polymorphisms.

niman – at 14:30

G1447A Again no match with other Qinghai strains, but easily found in other H5N1s

DQ659113 A/duck/Niger/914/2006 2006 H5N1

DQ320876 A/chicken/Fujian/1042/2005 2005 H5N1

DQ320875 A/duck/Fujian/897/2005 2005 H5N1

DQ320900 A/duck/Guangxi/951/2005 2005 H5N1

DQ320901 A/duck/Guangzhou/20/2005 2005 H5N1

DQ320911 A/duck/Hunan/1265/2005 2005 H5N1

DQ320912 A/duck/Hunan/1608/2005 2005 H5N1

DQ320913 A/duck/Hunan/1652/2005 2005 H5N1

DQ320939 A/duck/Vietnam/568/2005 2005 H5N1

DQ320896 A/goose/Guangxi/345/2005 2005 H5N1

DQ095628 A/Goose/Shantou/1621/05 2005 H5N1

DQ320915 A/goose/Shantou/2216/2005 2005 H5N1

DQ320916 A/migratory duck/Jiangxi/1653/2005 2005 H5N1

DQ320917 A/migratory duck/Jiangxi/1657/2005 2005 H5N1

DQ320918 A/migratory duck/Jiangxi/1701/2005 2005 H5N1

ISDN117777 A/Viet Nam/JP4207/2005 2005 H5N1

DQ320894 A/chicken/Guangxi/2448/2004 2004 H5N1

DQ320895 A/chicken/Guangxi/2461/2004 2004 H5N1

ISDN48972 A/Chicken/Hu bei/14/2004 2004 H5N1

AY684706 A/chicken/Hubei/327/2004 2004 H5N1

AY770079 A/chicken/Hubei/489/2004 2004 H5N1

AY830774 A/chicken/Macheng/2004 2004 H5N1

DQ083567 A/Ostrich/Samut Prakan/Thailand/CU-19/04 2004 H5N1

DQ083574 A/ostrich/Samut Prakan/Thailand/CU-31/04 2004 H5N1

AY651362 A/peregrine falcon/Hong Kong/D0028/2004 2004 H5N1

AY950236 A/swan/Guangxi/307/2004 2004 H5N1

AY651354 A/Ck/Hong Kong/NT93/2003 2003 H5N1

AY651356 A/Ck/Hong Kong/SSP141/2003 2003 H5N1

AY651355 A/Ck/Hong Kong/WF157/2003 2003 H5N1

niman – at 14:37

T1603 matches the low path ducks from Japan and a series of Qinghai isolates from Astrakhan but no match with Nigeria.

G253A has too many matches to list (several hundred H5N1’s) but no sharing wth any other Qinghai strains.

DQ659113 A/duck/Niger/914/2006 2006 H5N1

DQ389158 A/Cygnus olor/Astrakhan/Ast05–2−1/2005 2005 H5N1

DQ434889 A/Cygnus olor/Astrakhan/Ast05–2−10/2005 2005 H5N1

DQ343502 A/Cygnus olor/Astrakhan/Ast05–2−2/2005 2005 H5N1

DQ358746 A/Cygnus olor/Astrakhan/Ast05–2−3/2005 2005 H5N1

DQ363918 A/Cygnus olor/Astrakhan/Ast05–2−4/2005 2005 H5N1

DQ365004 A/Cygnus olor/Astrakhan/Ast05–2−5/2005 2005 H5N1

DQ364996 A/Cygnus olor/Astrakhan/Ast05–2−6/2005 2005 H5N1

DQ363923 A/Cygnus olor/Astrakhan/Ast05–2−7/2005 2005 H5N1

DQ399540 A/Cygnus olor/Astrakhan/Ast05–2−8/2005 2005 H5N1

DQ399547 A/Cygnus olor/Astrakhan/Ast05–2−9/2005 2005 H5N1

DQ449640 A/duck/Kurgan/08/2005 2005 H5N1

DQ320137 A/swan/Astrakhan/1/2005 2005 H5N1

AB241624 A/duck/Hokkaido/101/04 2004 H5N3

AB241625 A/duck/Hokkaido/193/04 2004 H5N3

AB241626 A/duck/Hokkaido/299/04 2004 H5N3

niman – at 15:00

Here is earlier commentary of the Nigerian sequence. It also had 6 “informative” polymorphisms, but they all matched other Qinghai isolates - 4 with Kurgan and 2 with Mongolia.

wetDirt – at 15:50

Dr Niman,

Your argument is based on the idea that these polymorphisms are not random, and that they are persistent across time and space. It seems to me that your data on regions of identity has gotten a lot of notice (admittedly, some tin-hat notice) because it directly supports your case that random mutations are rare, not common, in currently circulating strains of influenza A, both low and high path. Perhaps you could dig out some other examples of identical sequences, perhaps encompassing a few polymorphims, in low-path as well as hi-path strains. A region of identity in a low-path north American strain that persists for five or more years would be a powerful argument against the tin-foil-hatters, and strenghthen your argument that these regions are highly conserved.

And also. If you get Science, what do you think of the article from mid-February, vol 311 page 960, using the argument that recombination is an error-correcting mechanism. I though immediately of you when I read it.

26 July 2006

niman – at 09:50

The recent sequence from the Ivory Coast represents yet another introduction of H5N1 in western Africa (and more recombination).

The filed PCT has some comments on cached sequences correcting errors.

23 August 2006

moeb CFR6248at 19:37

yer a tricky devil mr. DemFromCt :-)

DemFromCTat 19:55

spam is spam and must be assimilated.

24 October 2006

Closed - Bronco Bill – at 22:00

Closed to maintain Forum speed.

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