Aggregation of Single Nucleotide Polymorphisms in a Human H5N1 Clade 2.2
Hemagglutinin
H. L. Niman
1
, M. D. Saad
2
, M. M. Aly
3
, J. A.Tjaden
2
, K. C. Earhart
2
, M. R.
Monteville
2
, M. M Mansour
2
, N. M Elsayed
5
, A. E. Nayel
5
, A. S. Abdelghani
5
, H.
M. Esmat
2, 5
, E. M. Labib
2
, E. A. Ayoub
2
, A. Arafa
3
, G. A. Raczniak
4.7
, M. Agyen-
Frempong
6
, W. K. Ampofo
7
, B. R. Boynton
2
1
Recombinomics, Inc., Pittsburgh, Pennsylvania, USA,
2
U.S. Naval Medical
Research Unit 3 (NAMRU-3), Cairo, Egypt,
3
Central Laboratory for Veterinary
Quality Control, Giza, Egypt,
4
NAMRU-3 Ghana Detachment, Accra, Ghana,
5
Ministry of Health, Arabic Republic of Egypt, Cairo, Egypt,
6
Ghana Veterinary
Services, Accra, Ghana,
7
Noguchi Memorial Institute for Medical Research,
Accra, Ghana
Correspondence to Henry L. Niman
Department of Influenza Recombination, Recombinomics, Inc, 648 Field Club
Road, Pittsburgh, PA 15238, USA
E-mail:
henry_niman@recombinomics.com
617.877.0987, 412.963.1362 FAX
Summary: Aggregation of single nucleotide polymorphisms challenges a basic
tenet of viral evolution, de novo random mutations.
Abstract word count: 108
Text word count: 1391
2 Figures 1 Supplemental Table
Running Title: Influenza SNP Aggregation
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
Abstract
The rapid evolution of the H5N1 serotype of avian influenza has been explained
by a mechanism involving the selection of single nucleotide polymorphisms
generated by copy errors. The recent emergence of H5N1 Clade 2.2 in fifty
countries, offered a unique opportunity to view the acquisition of new
polymorphism in these evolving genomes. We analyzed the H5N1 hemagglutinin
gene from a fatal human case from Nigeria in 2007. The newly emerged
polymorphisms were present in diverse H5N1 isolates from the previous year.
The aggregation of these polymorphisms from clade 2.2 sub-clades was not
supported by recent random mutations, and was most easily explained by
recombination between closely related sequences.
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
The evolution of H5N1 has attracted significant interest (1-4) due to linkages with
avian (5,6) and human infections (7,8). The basic tenets of influenza genetics (9)
attribute genetic drift to replication errors caused by a polymerase complex that
lacks a proof reading function. However, recent analysis (10) of swine influenza
genes identifies regions copied with absolute fidelity for more than 25 years. In
addition, polymorphism tracing of clade 2.2 H5N1 single nucleotide
polymorphisms identify concurrent acquisition (11) of the same polymorphism
onto multiple genetic backgrounds in widely dispersed geographical locations.
Here we show the aggregation of regional clade 2.2 polymorphisms from
Germany, Egypt, and sub-Sahara Africa onto a human Nigerian H5N1
hemagglutinin (HA), implicating recombination in the dispersal and aggregation
of single nucleotide polymorphisms from closely related genomes.
The rapid expansion of the geographical reach of H5N1 clade 2.2 has increased
attention on the mechanism of evolution in these rapidly changing genomes.
Clade 2.2 was first reported (12,13) at Qinghai Lake in Central China in May,
2005. Infection of long range migratory bird led to a rapid expansion of reported
H5N1 infections in wild birds and poultry in Europe, the Middle East, and Africa.
Moreover, human H5N1 infections have been subsequently reported in Turkey,
Iraq, Azerbaijan, Egypt, Djibouti, and Nigeria (14). More than 50 countries west
of China have reported clade 2.2 infections following the Qinghai Lake outbreak.
The expansion into these new regions was associated with the acquisition of
regional polymorphisms. Tracking of these newly acquired polymorphisms
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
demonstrates (11) a non-random appending of the polymorphisms on clade 2.2
genetic backgrounds. One NA polymorphism, G743A, which was a regional
polymorphism in Germany in 2006, was appended onto six distinct clade 2.2
genetic backgrounds in Russia, Egypt, and Ghana.
The HA phylogram in Figure 1A is annotated with regional polymorphism on
isolates from Egypt or neighboring countries (Israel, Gaza, and Djibouti). We
have designated these isolates as clade 2.2.1. The regional markers were
present in isolates from early 2006, but were also present in more recent
2006/2007 isolates from Egypt. Two of the polymorphisms, G467A and T937C
were present in all of the isolates from the region. Additional polymorphisms,
C661T, C727T, A880G, G1018T, were also in these regional isolates, but also
extended upstream to isolates in Europe, and downstream to isolates in Nigeria.
In addition to these regional markers, the figure identifies some of the sub-
regional markers, which will be discussed in more detail elsewhere.
The HA phylogram in Figure 1B is annotated with regional markers on isolates
from Germany and neighboring countries. The German isolates fell into three
major sub-clades. Isolates from northern Germany were similar to isolates from
Denmark (designated clade 2.2.2.1), and had sub-regional markers G1235A,
T1510C, and T1614C. A second sub-group (clade 2.2.2.2) has the broader
Egyptian markers, as well as G142A and A658B. A third sub-group (clade
2.2.2.3), has another series of markers (G41A, G295A, C689T, C1012T,
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
C1177T, C1402T, C1480T). Clade 2.2.2.3 has additional markers in NA,
including G743A, which was appended onto four different Egyptian backgrounds
in 2007 (11). Additional polymorphisms shared between German and Egyptian
isolates will be discussed in detail elsewhere.
The HA phylogram in Figure 1C is annotated with regional markers on isolates
from sub-Sahara Africa. These isolates also fall into three major sub-clades.
One group (clade 2.2.3.1) has the broader Egyptian markers plus G496A,
C627A, G1672A. A second group (clade 2.2.3.2) has G265A and T1614C. A
larger sub-Sahara group has A433G, G643A, and A1708G. The isolates from
Ghana had a number of additional polymorphisms appended onto this
background. The polymorphisms on the 2007 isolates from Ghana will be
discussed in detail elsewhere.
The first confirmed human clade 2.2 infection in Nigeria was in February, 2007.
The HA phylogram with this isolate and isolates which share polymorphisms with
the Nigerian sequence are listed in the phylogram in figure 2. The Nigerian
isolate has aggregated regional and sub-regional clade 2.2 polymorphisms from
Egypt (clade 2.2.1) Germany (clade 2.2.2.3), and sub-Sahara Africa. The isolate
has the three sub-Sahara polymorphisms, plus a sub-regional polymorphism,
A1006G from this sub-clade. This isolate also has a sub-set of the German
clade 2.2.2.3 markers (G295A and C1480T) as well as a sub-regional marker
from Egyptian clade 2.2.2.3, C1614T. In additional, the isolate has T937C, which
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
is one of the clade 2.2.1 regional markers. The human sequence also has two
clade 2.2.1 sub-regional markers (T610C and G643A). In addition, the sequence
has additional sub-regional markers from Mongolia and Siberia. Thus, 13 of the
16 newly acquired polymorphisms are present in sequences from clade 2.2
isolates, and six of the polymorphisms are regional markers.
Earlier reports have used phylogenetic analysis to conclude that H5N1 infections
in Nigeria involved multiple introductions (15-18). Similar observations have
been made for Germany (19-21). The similarities between isolates from Egypt
and Israel and Gaza have also been noted (22). The data presented here
support those conclusions, but the tracing of polymorphisms identifies exchanges
of polymorphisms between the sub-clades identified by this analysis. These
exchanges are not easily explained by random mutations due to copy errors.
Currently, genetic drift in influenza is explained by random mutations that
became dominant because of selection / adaptation pressures. However,
analysis of recent swine influenza sequences identified large regions of
nucleotide identity with sequences for isolates collected over 25 years earlier.
These isolate also have clear examples of homologous recombination, based on
matches with multiple parental sequences (10).
Recombination also offers an explanation for the aggregation data, which are not
easily explained by random mutations. The number of regional markers is small,
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
and the aggregation of subsets of polymorphisms into a single sequence does
not appear to be random. The aggregation data compliments the polymorphism
dispersal data (11) noted for NA G743A. The polymorphisms are appended onto
regional genetic backgrounds, but the new acquisitions have linkages to earlier
sequences that lie along migration pathways.
The acquisitions are most easily explained by recombination between closely
related sequences. A series of closely related sequences has been found in the
H5N1 reported in countries west of China. All of the reported cases have been
clade 2.2, and isolates are closely related to each other. However, the
sequences, as seen in the Egyptian isolates, are becoming more genetically
complex. As the sequence database grows, the ability to find matching
polymorphisms increases.
Theoretically, homologous recombination between closely related sequences
would be more common because of extensive regions of sequence identity.
Moreover, such recombination would result in acquisitions of single nucleotide
polymorphisms because most of the acquired sequences would be identical in
both parental and progeny sequences. Examples of closely related sequences in
human H5N1 have been reported previously in isolates with different receptor
binding domains (23,24), as well as susceptibility to anti-viral treatment with
oseltamivir (25). Plaques purified clones from the same patient had different
combinations of receptor binding domain polymorphisms. Similarly, plaque
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
purified clones with two different oseltamivir resistance changes, H274Y and
N294S, were isolated, in addition to clones with wild type polymorphisms.
However, both of these changes have been detected in poultry or wild bird
sequences that have not been linked to oseltamivir sequences, raising the
possibility that the resistance markers that emerged were already present in low
abundance prior to treatment.
One of the markers, N294S, was also in clade 2.2 sequences present in family
members from the Egyptian governorate of Gharbiya. This change was present
in isolates collected prior to treatment, as well as isolates from samples collected
two days following the start of treatment. Sequences from two of the patients
were similar but distinct, and these distinctions were present in both sets of
sequences, suggesting that the source of infection for the patients also contained
distinct but closely related sequences.
Similarly, plaque purified clones of H5N1 from a chicken in Gharbiya identified
two distinct populations. One was closely related to the sequences from the
patients in Gharbiya, while the other was identical to sequences from other
chickens in Gharbiya. Moreover, the clones had evidence of recombination (data
will be reported elsewhere). Examples of dual infections involving closely related
sequences (26) provide the genetic basis for acquisition of single nucleotide
polymorphisms by recombination.
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
Tracing of these polymorphisms defines distribution routes. The distribution of
additional polymorphisms described above will be detailed elsewhere. The
mapping of these polymorphisms has the potential of defining predictable
acquisitions. The predicted acquisitions can then be used to create vaccine
targets representing emerging genomes.
Figure legends
Figure 1 HA Phylograms of Egyptian, German, and Sub-Sahara Isolates
A. Phylogram of Egyptian and regional isolates. Nucleotide number corresponds
to HA sequence from cat/Germany/606/2006. Clade 2.2.1 Egyptian regional
markers are G467A and T937C. Extended markers are C661T, C727T, A880G,
G1018T.
B. Phylogram of German and regional isolates. Clade 2.2.2.1 German regional
markers are G1235A and T1510C. Extended marker is T1614C. Clade 2.2.2.2
German regional markers are G142A and A658G. Extended markers are
C661T, C727T, A880G, G1018T. Clade 2.2.2.3 German regional markers are
G41A, G295A, C689T, C1012T, C1177T, C1402T, C1480T.
C. Phylogram of sub-Sahara isolates. Sub-Sahara markers are A433G, G643A,
A1708G. Clade 2.2.3.1 Nigeria regional markers are G496A, C627A, G1672A.
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
Extended markers are C661T, C727T, A880G, G1018T. Clade 2.2.3.2 Nigeria
regional markers are G265A and T1614C.
Phylograms represent positions 93-1688. Isolates and accession numbers are in
Table S1. Trees were generated using neighbor joining with 100 bootstrap
repetitions. Sequences generated as described previously (27)
Figure 2. Aggregated Polymorphisms on Human Nigerian Hemagglutinin
Phylogram as described in Figure 1. 13 of the 16 polymorphisms are found in
other clade 2.2 isolates including Egyptian regional polymorphism T937C
(acqua), German clade 2.2.2.3 regional polymorphisms G295A (red) and C1480T
(dark red), and sub-Sahara regional polymorphisms A433G (light orange),
G643A (violet), A1708G (pink). Sub-regional polymorphisms include C289T (sea
green), T610C (dark gray), A778G (orange), G781A (light blue), C981T (pale
blue), A1006G (light gray), G1658A (turquoise). Polymorphisms not found in
other clade 2.2 sequences are G167A, G1005A, G1430A.
Acknowledgements
We thank Cecilia DeMattos from US NAMRU-3 for plaque purification of avian
isolates; Evelyn Yayra Bonney, Ivy Asante, Ken-Edwin Aryee, Kofi Bonney,
Jacob Barnor and Professor Alexander Nyarko from Noguchi Memorial Institute
for Medical Research; Mr. Jerry Odoi, Drs. Joseph Gaari-Kweku, Joseph
Adongo Awuni, John Niendow Karimu and Bashiru Kikimoto from the Ghana
Veterinary Services; Mr. Michael Adjabeng, Drs. Edward Antwi and Lawson
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
Ahadzie from the Ghana Health Service; for the collection, screening and testing
of avian and human samples for H5N1 in Ghana.
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Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
Figure 1A. Phylogram of Egyptian and regional isolates
C661T
C727T
A880G
G1018T
G1018T
A880G
C727T
C661T
G467A
T937C
G467A
T937C
C689T
C689T
G754A
A356G
3 BP del
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
Figure 1B. Phylogram of German and regional isolates
G41A
G295A
C689T
C689T
C1012T
C1177T
C1402T
C1480T
C1480T
C1141T
G516T
C1282T
C1141T
G1235A
T1510C
T1614C
G41A
G41A
G295A
C1012T
C1177T
C1402T
C1282T
A1612G
G142A
A658G
C661T
C727T
A880G
G1018T
C796T
G295A
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
Figure 1C. Phylogram of sub-Sahara isolates
T154C
G265A
G265A
T1614C
T1614C
C427T
C1210T
G1439A
C206T
A539G
C599T
A855G
C895T
C970T
T1359C
C1432T
A433G
G643A
A1708G
G535A
A1684C
A1006G
G1679A
C627A
G1672A
C661T
G1018T
C727T
C1216T
A863G
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
Figure 2. Aggregated Polymorphisms in Human Nigerian Hemagglutinin
G295A
G295A
C289T
A433G
T610C
T610C
G643A
A1708G
T937C
T937C
A1006G
C1480T
C1480T
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007
Table S1 H5N1 Isolates and Accession Numbers
Accession
Name
Year
Figure
EF624253
A/chicken/Ghana/3158-NAMRU3/2007
2007
1C, 2
EF624254
A/chicken/Ghana/3159-NAMRU3/2007
2007
1C, 2
EF624255
A/chicken/Ghana/3160-NAMRU3/2007
2007
1C, 2
EF441277
A/chicken/Egypt/1079-NAMRU3/2007
2007
1A, 2
EF469650
A/chicken/Egypt/1129N3-HK9/2007
2007
1A, 2
EF441280
A/chicken/Egypt/1300-NAMRU3/2007
2007
1A, 2
EF469653
A/chicken/Egypt/1889N3-SM26/2007
2007
1A, 2
EF469654
A/chicken/Egypt/1890N3-HK45/2007
2007
1A, 2
EF469659
A/chicken/Egypt/1891N3-CLEVB/2007
2007
1A, 2
EF469660
A/chicken/Egypt/1892N3-HK49/2007
2007
1A, 2
EF441281
A/duck/Egypt/1301-NAMRU3/2007
2007
1A, 2
EF469657
A/duck/Egypt/1888N3-SM25/2007
2007
1A, 2
EF382359
A/Egypt/0636-NAMRU3/2007
2007
1A, 2
EF535817
A/Egypt/1394-NAMRU3/2007
2007
1A, 2
EF535818
A/Egypt/1604-NAMRU3/2007
2007
1A, 2
EF535819
A/Egypt/1731-NAMRU3/2007
2007
1A, 2
EF535820
A/Egypt/1902-NAMRU3/2007
2007
1A, 2
EF535821
A/Egypt/2256-NAMRU3/2007
2007
1A, 2
EF535822
A/Egypt/2321-NAMRU3/2007
2007
1A, 2
EF535823
A/Egypt/2331-NAMRU3/2007
2007
1A, 2
EF535824
A/Egypt/2616-NAMRU3/2007
2007
1A, 2
EF535825
A/Egypt/2620-NAMRU3/2007
2007
1A, 2
EF535826
A/Egypt/2621-NAMRU3/2007
2007
1A, 2
EU095025
A/Egypt/2629-NAMRU3/2007
2007
1A, 2
EU095026
A/Egypt/2630-NAMRU3/2007
2007
1A, 2
EU095027
A/Egypt/2631-NAMRU3/2007
2007
1A, 2
EU095028
A/Egypt/2750-NAMRU3/2007
2007
1A, 2
EU095029
A/Egypt/2751-NAMRU3/2007
2007
1A, 2
EU095030
A/Egypt/4081-NAMRU3/2007
2007
1A, 2
EU095031
A/Egypt/4082-NAMRU3/2007
2007
1A, 2
EU095032
A/Egypt/4226-NAMRU3/2007
2007
1A, 2
EF441276
A/chicken/Egypt/1078-NAMRU3/2006
2006
1A, 2
EF441278
A/chicken/Egypt/1080-NAMRU3/2006
2006
1A, 2
EF441279
A/chicken/Egypt/1081-NAMRU3/2006
2006
1A, 2
EF042622
A/chicken/Egypt/10845-NAMRU3/2006
2006
1A, 2
EF469651
A/chicken/Egypt/12378N3-CLEVB/2006
2006
1A, 2
EF469652
A/chicken/Egypt/12379N3-CLEVB/2006
2006
1A, 2
DQ837587
A/chicken/Egypt/5610NAMRU3-F3/2006
2006
1A, 2
DQ837588
A/chicken/Egypt/5611NAMRU3-AN/2006
2006
1A, 2
DQ837589
A/chicken/Egypt/5612NAMRU3-S/2006
2006
1A, 2
DQ447199
A/chicken/Egypt/960N3-004/2006
2006
1A, 2
EF469655
A/duck/Egypt/12380N3-CLEVB/2006
2006
1A, 2
EF469656
A/duck/Egypt/13010N3-CLEVB/2006
2006
1A, 2
EF061116
A/Egypt/12374-NAMRU3/2006
2006
1A, 2
EF200512
A/Egypt/14724-NAMRU3/2006
2006
1A, 2
EF200513
A/Egypt/14725-NAMRU3/2006
2006
1A, 2
EF042614
A/Egypt/2763-NAMRU3/2006
2006
1A, 2
DQ464377
A/Egypt/2782-NAMRU3/2006
2006
1A, 2
EF042615
A/Egypt/2783-NAMRU3/2006
2006
1A, 2
EF042616
A/Egypt/2786-NAMRU3/2006
2006
1A, 2
EF042617
A/Egypt/2947-NAMRU3/2006
2006
1A, 2
EF042618
A/Egypt/3105-NAMRU3/2006
2006
1A, 2
EF042619
A/Egypt/3458-NAMRU3/2006
2006
1A, 2
EF042620
A/Egypt/5494-NAMRU3/2006
2006
1A, 2
EF042621
A/Egypt/5614-NAMRU3/2006
2006
1A, 2
EF469658
A/goose/Egypt/13009N3-SM2/2006
2006
1A, 2
DQ837590
A/turkey/Egypt/5613NAMRU3-T/2006
2006
1A, 2
EF042624
A/teal/Egypt/14051-NAMRU3/2005
2005
1A, 1B, 2
EF474450
A/chicken/Moscow/2/2007
2007
1B, 2
EF605603
A/chicken/Russia_Krasnodar/2/2007
2007
1B, 2
ISDN230945 A/Nigeria/6e/07
2007
1C, 2
DQ822563
A/bar-headed goose/Qinghai/F/2006
2006
2
DQ822564
A/black-headed gull/Qinghai/3/2006
2006
2
EF165057
A/buzzard/Bavaria/13/2006
2006
1A,1B, 2
EF165049
A/buzzard/Bavaria/5/2006
2006
1A,1B, 2
EF523687
A/buzzard/Denmark/6370/06
2006
1A,1B, 2
AM403474
A/Canada goose/Germany/R1207/06
2006
1A,1B, 2
AM403461
A/Canada goose/Germany/R71/06
2006
1A,1B, 2
EF395844
A/cat/Austria/649/2006
2006
1B, 2
DQ643982
A/cat/Germany/606/2006
2006
1A,1B, 2
AM403468
A/cat/Germany/R606/06
2006
1A,1B, 2
AM400974
A/chicken/Burkina Faso/01.03/2006
2006
1C, 2
AM400973
A/chicken/Burkina Faso/13.1/2006
2006
1C, 2
CY016811
A/chicken/Cote d'Ivoire/1787-34/2006
2006
1C, 2
EF532628
A/chicken/Gaza/450/2006
2006
1A, 2
EF532630
A/chicken/Gaza/713/2006
2006
1A, 2
EF532631
A/chicken/Gaza/714/2006
2006
1A, 2
EF532626
A/chicken/Israel/397/2006
2006
1A, 2
EF532629
A/chicken/Israel/625/2006
2006
1A, 2
CY021517
A/chicken/Ivory Coast/1787-35/2006
2006
1C, 2
DQ673901
A/chicken/Ivory Coast/1787/2006
2006
1C, 2
DQ676834
A/chicken/Krasnodar/01/2006
2006
1B, 2
EF205159
A/chicken/Krasnodar/123/06
2006
1B, 2
DQ864718
A/chicken/Krasnodar/199/06
2006
1B, 2
AM262541
A/chicken/Lagos.NIE/BA209/2006
2006
1C, 2
AM262542
A/chicken/Lagos.NIE/BA210/2006
2006
1C, 2
AM262543
A/chicken/Lagos.NIE/BA211/2006
2006
1C, 2
AM262546
A/chicken/Lagos.NIE/SO300/2006
2006 1A, 1B, 1C, 2
AM262547
A/chicken/Lagos.NIE/SO452/2006
2006 1A, 1B, 1C, 2
AM262553
A/chicken/Lagos.NIE/SO493/2006
2006 1A, 1B, 1C, 2
AM262572
A/chicken/Lagos.NIE/SO494/2006
2006 1A, 1B, 1C, 2
DQ838517
A/chicken/Niger/2130-7/2006
2006
1C, 2
DQ838516
A/chicken/Niger/2130-8/2006
2006
1C, 2
CY016939
A/chicken/Nigeria/1047-30/2006
2006
1C, 2
CY016947
A/chicken/Nigeria/1047-34/2006
2006
1C, 2
CY016923
A/chicken/Nigeria/1047-54/2006
2006
1C, 2
CY016931
A/chicken/Nigeria/1047-62/2006
2006
1C, 2
CY016907
A/chicken/Nigeria/1047-8/2006
2006
1C, 2
CY016276
A/chicken/Nigeria/641/2006
2006
1C, 2
CY016284
A/chicken/Nigeria/957-20/2006
2006
1C, 2
AM503002
A/chicken/Nigeria/AB13/2006
2006
1C, 2
AM503003
A/chicken/Nigeria/AB14/2006
2006
1C, 2
AM503004
A/chicken/Nigeria/FA4/2006
2006
1C, 2
AM502998
A/chicken/Nigeria/FA6/2006
2006
1C, 2
AM502999
A/chicken/Nigeria/FA7/2006
2006
1C, 2
AM503001
A/chicken/Nigeria/IF10/2006
2006
1C, 2
AM503005
A/chicken/Nigeria/OD8/2006
2006
1C, 2
AM503000
A/chicken/Nigeria/OD9/2006
2006
1C, 2
AM400975
A/chicken/Nigeria/SO227/2006
2006
1C, 2
AM400976
A/chicken/Nigeria/SO294/2006
2006
1C, 2
AM400977
A/chicken/Nigeria/SO457/2006
2006
1C, 2
AM400978
A/chicken/Nigeria/SO461/2006
2006
1C, 2
AM400979
A/chicken/Nigeria/SO470/2006
2006
1C, 2
AM400980
A/chicken/Nigeria/SO478/2006
2006
1C, 2
AM400981
A/chicken/Nigeria/SO485/2006
2006
1C, 2
CY016300
A/chicken/Sudan/1784-10/2006
2006
1C, 2
CY016292
A/chicken/Sudan/1784-7/2006
2006
1C, 2
CY020661
A/chicken/Sudan/1784-8/2006
2006
1C, 2
DQ862003
A/chicken/Sudan/1784/2006
2006
1C, 2
CY021389
A/chicken/Sudan/2115-10/2006
2006
1C, 2
CY020677
A/chicken/Sudan/2115-12/2006
2006
1C, 2
CY020669
A/chicken/Sudan/2115-9/2006
2006
1C, 2
DQ864719
A/chicken/Volgograd/236/06
2006
1B, 2
DQ659679
A/common bussard/Bavaria/2/2006
2006
1A, 1B, 2
EF165055
A/common buzzard/Bavaria/11/2006
2006
1A, 1B, 2
AM403463
A/common buzzard/Germany/R306/06
2006
1A, 1B, 2
AM408213
A/common buzzard/Germany/R870/06
2006
1A, 1B, 2
EF110519
A/common coot/Switzerland/V544/06
2006
1A, 1B, 2
EF165051
A/common pochard/Bavaria/7/2006
2006
1A, 1B, 2
EU016355
A/common pochard/Switzerland/V505/2006
2006
1A, 1B, 2
EU016358
A/common pochard/Switzerland/V592/2006
2006
1A, 1B, 2
EU016359
A/common pochard/Switzerland/V762/2006
2006
1A, 1B, 2
AM403469
A/coot/Germany/R655/06
2006
1A, 1B, 2
AM408209
A/cormorant/Germany/R292/06
2006
1A, 1B, 2
CY016779
A/Cygnus cygnus/Iran/754/2006
2006
1B, 2
DQ515984
A/Cygnus olor/Czech Republic/5170/2006
2006
1A, 1B, 2
CY017035
A/Cygnus olor/Italy/742/2006
2006
1A, 1B, 2
CY020349
A/cygnus olor/Italy/808/2006
2006
1B, 2
CY016803
A/duck/Cote d'Ivoire/1787-18/2006
2006
1C, 2
EF532632
A/duck/Gaza/760/2006
2006
1A, 2
EF532622
A/duck/Gaza/834/2006
2006
1A, 2
AM403464
A/duck/Germany/R338/06
2006
1A, 1B, 2
AM403466
A/duck/Germany/R592/06
2006
1A, 1B, 2
AM403467
A/duck/Germany/R603/06
2006
1A, 1B, 2
AM403470
A/duck/Germany/R751/06
2006
1A, 1B, 2
CY017027
A/duck/Niger/914/2006
2006
1C, 2
EU016352
A/duck/Switzerland/V389/2006
2006
1A, 1B, 2
EU016353
A/duck/Switzerland/V426/2006
2006
1A, 1B, 2
EU016354
A/duck/Switzerland/V487/2006
2006
1A, 1B, 2
EF165054
A/eagle owl/Bavaria/10/2006
2006
1A, 1B, 2
AM403473
A/eagle owl/Germany/R1166/06
2006
1A, 1B, 2
EF165059
A/falcon/Bavaria/15/2006
2006
1A, 1B, 2
AM408211
A/falcon/Germany/R899/06
2006
1A, 1B, 2
EF523696
A/fowl/Denmark/60296/06
2006
1A, 1B, 2
EF165063
A/goldeneye duck/Bavaria/19/2006
2006
1A, 1B, 2
EF165062
A/goosander/Bavaria/18/2006
2006
1A, 1B, 2
EF165064
A/goosander/Bavaria/20/2006
2006
1A, 1B, 2
EF110518
A/goosander/Switzerland/V82/06
2006
1A, 1B, 2
EF165066
A/great crested grebe/Bavaria/22/2006
2006
1A, 1B, 2
EF523695
A/great crested grebe/Denmark/7498/06
2006
1A, 1B, 2
AM408212
A/great crested grebe/Germany/R1226/06
2006
1A, 1B, 2
EF523688
A/grey lag goose/Denmark/6692/06
2006
1B, 2
CY017179
A/guinea fowl/Nigeria/957-12/2006
2006
1C, 2
AM408215
A/gull/Germany/R882/06
2006
1A, 1B, 2
AM400971
A/hooded vulture/Burkina Faso/1/2006
2006
1C, 2
AM400972
A/hooded vulture/Burkina Faso/2/2006
2006
1C, 2
EU016351
A/little grebe/Switzerland/V330/2006
2006
1A, 1B, 2
DQ458992
A/mallard/Bavaria/1/2006
2006
1A, 1B, 2
CY016795
A/mallard/Italy/835/2006
2006
1A, 1B, 2
EU016356
A/mallard/Switzerland/V537/2006
2006
1A, 1B, 2
EU016357
A/mallard/Switzerland/V558/2006
2006
1A, 1B, 2
EF165056
A/mute swan/Bavaria/12/2006
2006
1A, 1B, 2
AM403460
A/mute swan/Germany/R65/06
2006
1A, 1B, 2
EF547198
A/mute swan/Germany/R797/06
2006
1A, 1B, 2
AM403471
A/mute swan/Germany/R854/06
2006
1A, 1B, 2
EU016350
A/mute swan/Switzerland/V68/2006
2006
1A, 1B, 2
CY016915
A/ostrich/Nigeria/1047-25/2006
2006
1C, 2
EF523689
A/peacock/Denmark/60295/06
2006
1A, 1B, 2
EF523690
A/peregrine/Denmark/6632/06
2006
1A, 1B, 2
AM403465
A/pochard/Germany/R348/06
2006
1A, 1B, 2
AM492165
A/stone marten/Germany/R747/2006
2006
1A, 1B, 2
AM403475
A/stork/Germany/R1239/06
2006
1A, 1B, 2
EF395845
A/swan/Austria/216/2006
2006
1A, 1B, 2
EF165058
A/swan/Bavaria/14/2006
2006
1A, 1B, 2
EF165060
A/swan/Bavaria/16/2006
2006
1A, 1B, 2
EF165061
A/swan/Bavaria/17/2006
2006
1A, 1B, 2
EF165065
A/swan/Bavaria/21/2006
2006
1A, 1B, 2
EF165050
A/swan/Bavaria/6/2006
2006
1A, 1B, 2
DQ464354
A/swan/Germany/R65/2006
2006
1A, 1B, 2
DQ440535
A/swan/Iran/754/2006
2006
1B,2
DQ822565
A/swan/Qinghai/01/2006
2006
2
CY017043
A/swan/Slovenia/760/2006
2006
1A, 1B, 2
EF165048
A/tufted duck/Bavaria/4/2006
2006
1A, 1B, 2
EF165052
A/tufted duck/Bavaria/8/2006
2006
1A, 1B, 2
EF165053
A/tufted duck/Bavaria/9/2006
2006
1A, 1B, 2
EF523691
A/tufted duck/Denmark/6431/06
2006
1A, 1B, 2
EF523692
A/tufted duck/Denmark/6540/06
2006
1A, 1B, 2
AM408216
A/tufted duck/Germany/R1240/06
2006
1A, 1B, 2
EF547197
A/tufted duck/Switzerland/V504/06
2006
1A, 1B, 2
EF619982
A/Turkey/12/2006
2006
1B, 2
EF619989
A/Turkey/15/2006
2006
1B, 2
EF619990
A/Turkey/651242/2006
2006
1B, 2
EF619998
A/Turkey/65596/2006
2006
1B, 2
EF532623
A/turkey/Israel/345/2006
2006
1A, 2
EF532624
A/turkey/Israel/364/2006
2006
1A, 2
EF532625
A/turkey/Israel/365/2006
2006
1A, 2
EF532627
A/turkey/Israel/446/2006
2006
1A, 2
CY020693
A/turkey/Ivory Coast/4372-2/2006
2006
1A, 2
CY020701
A/turkey/Ivory Coast/4372-3/2006
2006
1A, 2
CY020709
A/turkey/Ivory Coast/4372-4/2006
2006
1A, 2
EF523693
A/whooper swan/Denmark/7224/06
2006
1A, 1B, 2
EF523694
A/whooper swan/Denmark/7275/06
2006
1A, 1B, 2
AM403462
A/whooper swan/Germany/R88/06
2006
1A, 1B, 2
DQ650659
A/chicken/Crimea/04/2005
2005
1B, 2
DQ650663
A/chicken/Crimea/08/2005
2005
1B, 2
DQ449632
A/chicken/Kurgan/05/2005
2005
2
DQ323672
Achicken/Kurgan/3/2006
2005
2
EF205154
A/chicken/Suzdalka/06/05
2005
2
DQ389158
A/Cygnus olor/Astrakhan/Ast05-2-1/2005
2005
1B, 2
DQ434889
A/Cygnus olor/Astrakhan/Ast05-2-10/2005
2005
1B, 2
DQ343502
A/Cygnus olor/Astrakhan/Ast05-2-2/2005
2005
1B, 2
DQ358746
A/Cygnus olor/Astrakhan/Ast05-2-3/2005
2005
1B, 2
DQ363918
A/Cygnus olor/Astrakhan/Ast05-2-4/2005
2005
1B, 2
DQ365004
A/Cygnus olor/Astrakhan/Ast05-2-5/2005
2005
1B, 2
DQ364996
A/Cygnus olor/Astrakhan/Ast05-2-6/2005
2005
1B, 2
DQ363923
A/Cygnus olor/Astrakhan/Ast05-2-7/2005
2005
1B, 2
DQ399540
A/Cygnus olor/Astrakhan/Ast05-2-8/2005
2005
1B, 2
DQ399547
A/Cygnus olor/Astrakhan/Ast05-2-9/2005
2005
1B, 2
DQ449640
A/duck/Kurgan/08/2005
2005
1C, 2
DQ676840
A/goose/Krasnoozerka/627/2005
2005
1B, 2
EF205157
A/goose/Krasnoozerskoe/627/05
2005
1B, 2
EF205156
A/goose/Suzdalka/10/05
2005
2
EF205158
A/turkey/Suzdalka/12/05
2005
2
EF619980
A/turkey/Turkey/1/2005
2005
1B, 1C, 2
AB233322
A/whooper swan/Mongolia/6/05
2005
2
ISDN138113 A/whooper swan/Mongolia/7/2005
2005
2
Nature Precedings : hdl:10101/npre.2007.743.2 : Posted 10 Sep 2007