Aggregation of Single Nucleotide Polymorphisms in a Human H5N1 Clade 2.2
Hemagglutinin
Niman HL*, Saad MD
#
, Tjaden JA
#
, Earhart KC
#
, Monteville MR
#
, Aly MM
%
,
Mansour MM
#
, Elsayed NM
+
, Nayel AE
+
, Abdelghani AS
+
, Esmat HM
#+
, Labib
EM
#
, Ayoub EA
#
, Arafa A-SA
%
, Raczniak GA
##,####
, Agyen-Frempong M
###
,
Ampofo WK
####
, Boynton BR
#
*Recombinomics, Inc., Pittsburgh, Pennsylvania, USA,
#
U.S. Naval Medical
Research Unit 3 (NAMRU-3), Cairo, EGY,
##
NAMRU-3 Ghana Detachment,
Accra, GHA,
%
Central Laboratory for Veterinary Quality Control, Giza, EGY,
+
Ministry of Health, Arabic Republic of Egypt, Cairo, EGY,
###
Ghana Veterinary
Services, Accra, GHA,
####
Noguchi Memorial Institute for Medical Research,
Accra, GHA
Department of Influenza Recombination, Recombinomics, Inc, 648 Field Club
Road, Pittsburgh, PA 15238, USA
Correspondence to Henry L Niman, E-mail:
henry_niman@recombinomics.com
617.877.0987, 412.963.1362 FAX
Text word count: 1391
2 Figures 1 Supplemental Table
Running Title: H5N1 Polymorphism Aggregation
Nature Precedings : hdl:10101/npre.2007.743.1 : Posted 16 Aug 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.2007.743.1 : Posted 16 Aug 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, G1019A, 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 T1615C. 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.2007.743.1 : Posted 16 Aug 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 G209A and T1415C. 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 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 is one of the clade 2.2.1 regional
Nature Precedings : hdl:10101/npre.2007.743.1 : Posted 16 Aug 2007
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 14 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.2007.743.1 : Posted 16 Aug 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
, as well as susceptibility to anti-viral treatment with oseltamivir
24
. Plaques purified clones from the same patient had different combinations of
receptor binding domain polymorphisms. Similarly, plaque purified clones with
Nature Precedings : hdl:10101/npre.2007.743.1 : Posted 16 Aug 2007
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). These examples of dual infections involving closely
related sequences provide the genetic basis for acquisition of single nucleotide
polymorphisms by recombination.
Nature Precedings : hdl:10101/npre.2007.743.1 : Posted 16 Aug 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 with clade 2.2.1 Egyptian regional
markers G467A and T937C. Extended markers include C661T, C727T, A880G,
G1019T.
B. Phylogram of German and regional isolates. Clade 2.2.2.1 German regional
markers G1235A and T1510C, and extended marker T1615C. Clade 2.2.2.2
with German regional markers G142A and A658G. Extended markers include
C661T, C727T, A880G, G1019T. Clade 2.2.2.3 German regional markers 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 has Nigeria regional markers G496A, C627A, G1672A
and extended markers C661T, C727T, A880G, G1019T. Clade 2.2.3.2 has
Nigeria regional markers G209A and T1415C.
Nature Precedings : hdl:10101/npre.2007.743.1 : Posted 16 Aug 2007
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
25
Figure 2 Aggregated Polymorphisms on Human Nigerian Hemagglutinin
Phylogram as described in Figure 1. 13 of the 14 polymorphisms are found in
other clade 2.2 including Egyptian regional polymorphism T937C, German clade
2.2.2.3 regional polymorphisms G295A and C1480T, and sub-Sahara regional
polymorphisms A433G, G643A, G1708A.
Acknowledgements
We thank Evelyn Yayra Bonney, Ken-Edwin Aryee, Kofi Bonney, and Professor
Alexander Nyarko from Naguchi Memorial Institute for Medical Research, and
Drs Joseph GAARI-KWEKU, Joseph ADONGO AWUNI, John NIENDOW
KARIMU, Bashiru KIKIMOTO and Jerry ODOI from the Ghana Veterinary
Services
for the collection of avian samples.
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Figure 1A. Phylogram of Egyptian and regional isolates
Nature Precedings : hdl:10101/npre.2007.743.1 : Posted 16 Aug 2007
Figure 1B. Phylogram of German and regional isolates
Nature Precedings : hdl:10101/npre.2007.743.1 : Posted 16 Aug 2007
Figure 1C. Phylogram of sub-Sahara isolates
Nature Precedings : hdl:10101/npre.2007.743.1 : Posted 16 Aug 2007
Figure 2. Aggregated Polymorphisms on Human Nigerian Hemagglutinin
Nature Precedings : hdl:10101/npre.2007.743.1 : Posted 16 Aug 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
A/Egypt/2629-NAMRU3/2007
2007
1A, 2
A/Egypt/2630-NAMRU3/2007
2007
1A, 2
A/Egypt/2631-NAMRU3/2007
2007
1A, 2
A/Egypt/2750-NAMRU3/2007
2007
1A, 2
A/Egypt/2751-NAMRU3/2007
2007
1A, 2
A/Egypt/4081-NAMRU3/2007
2007
1A, 2
A/Egypt/4082-NAMRU3/2007
2007
1A, 2
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
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.2007.743.1 : Posted 16 Aug 2007