Phylogenetic Analysis of Austrian Canine Distemper Virus From Clinical Samples From Dogs and Wild...

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Phylogenetic analysis of Austrian canine

distemper virus strains from clinical samplesfrom dogs and wild carnivores

V. Benetka, M. Leschnik, N. Affenzeller, K. Mstl

Austrian field cases of canine distemper (14 dogs, one badger [Meles meles] and onestone marten [Martes foina]) from 2002 to 2007 were investigated and the case historieswere summarised briefly. Phylogenetic analysis of fusion (F) and haemagglutinin (H) genesequences revealed different canine distemper virus (CDV) lineages circulating in Austria.

The majority of CDV strains detected from 2002 to 2004 were well embedded in the Europeanlineage. One Austrian canine sample detected in 2003, with a high similarity to Hungariansequences from 2005 to 2006, could be assigned to the Arctic group (phocine distemper virustype 2-like). The two canine sequences from 2007 formed a clearly distinct group flanked bysequences detected previously in China and the USA on an intermediate position betweenthe European wildlife and the Asia-1 cluster. The Austrian wildlife strains (2006 and 2007)could be assigned to the European wildlife group and were most closely related to, yet clearlydifferent from, the 2007 canine samples. To elucidate the epidemiological role of Austrianwildlife in the transmission of the disease to dogs and vice versa, H protein residues relatedto receptor and host specificity (residues 530 and 549) were analysed. All samples showedthe amino acids expected for their host of origin, with the exception of a canine sequencefrom 2007, which had an intermediate position between wildlife and canine viral strains.

In the period investigated, canine strains circulating in Austria could be assigned to fourdifferent lineages reflecting both a high diversity and probably different origins of virusintroduction to Austria in different years.

CANINE distemper is a widespread, multisystemic, often fatal dis-ease of dogs and other species of Canidae, Mustelidae, Procyonidae,Ursidae and the large Felidae. A similar disease, which affects pinni-peds and is known as phocine distemper, is caused by phocine dis-temper virus (PDV). Viraemia often leads to the invasion of the CNS,and the outcome in animals with severe clinical CNS signs is nearlyalways fatal (Greene and Appel 2006). Canine distemper virus (CDV)is a single-stranded RNA virus in the order Mononegavirales, familyParamyxoviridae and genusMorbillivirus. The complete genome con-sists of 15,690 bases encoding seven proteins (Sidhu and others 1993).The fusion (F) protein is an integral membrane protein binding cellsurface receptors of target cells. The F protein and the haemaggluti-

V. Benetka,DVM,K. Mstl,DVM,Clinical Virology, Department for

Pathobiology,M. Leschnik, DVM,

N. Affenzeller,DVM,

Clinic of Internal Medicine and InfectiousDiseases,

Department for Companion Animalsand Horses, University of Veterinary

Medicine, Veterinaerplatz 1, A-1210Vienna, Austria

Correspondence to Dr Mstl, e-mail:

[email protected]

Provenance: not commissioned;externally peer reviewed

Published Online First April 6, 2011

Veterinary Record (2011) 168, 377 doi: 10.1136/vr.c6404

nin (H) protein are both envelope proteins involved in viral-inducedcell-to-cell fusion, cytopathogenicity and cell tropism. When thereis interchange of H proteins between different strains of CDV andthe measles virus, and of both F and H proteins between highly andless pathogenic CDV strains, recombinant strains display the char-acteristics of the original donator strains, showing higher or lowercytopathogenicity in cell culture and virulence in the animal modelthan their non-recombinant counterparts (von Messling and others2001, 2003).

Sequence analyses of the variable H genes of field isolates haverevealed different lineages of CDV strains circulating worldwide (Boltand others 1997, Haas and others 1997, Iwatsuki and others 1997,Pardo and others 2005, Martella and others 2006, Demeter and others2007): lineage USA-1, with the vaccine strains Convac, Onderstepoort,Snyder-Hill; lineage USA-2, with US isolates from different hosts (canineand wildlife) closely related to each other yet distinct from the vaccinestrains; the Asian cluster (Asia-1 and Asia-2); and the European cluster,consisting of the European wildlife group and the so-called Arctic line-age formed by sequences similar to PDV-2. A cluster of sequences fromthe USA and China (AF178038, red panda [Ailurus fulgens]; AF178039,giant panda [Ailuropoda meanoleuca]; and AY964114, dog) occupies anintermediate position between the Asia-1 and the European wildlifegroup. Their classification into one of these two groups varies between

the different genomic regions investigated and the authors who con-ducted the research (Martella and others 2006, McCarthy and others2007, An and others 2008, Han and others 2008). Arctic-like sequenceshave been detected in dogs in Italy and Hungary. Therefore, overall, theclustering of CDV isolates seems to be more related to geographic thanhost origin (Harder and others 1996, Zhao and others 2010). However,

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was carried out in a volume of 20 l with a ready to use sequencingPCR mixture (DNA Sequencing Kit; Applied Biosystems); for eachsample, the forward and reverse sequences were determined.

Three sections of the viral genome were aligned and used for phylo-genetic analyses (bp positions relative to the strain Onderstepoort;GenBank accession number AF378705): F gene segment 1, position5200-5980, 781 bp; F gene segment 2, position 6284-6923, 640 bp;and H gene segment 1, position 7079-7878, 800 bp. For a selection of

available samples (Fig 1), the complete H gene, position 7079-8902,1824 bp was used.For alignment of nucleotide and amino acid sequences, the pro-

gram Align Plus 4 (Scientific and Educational Software) and the freeClustalX (www.clustal.org) software were used. Phylogenetic treeswere constructed using the program MEGA 4.1 (www.megasoftware.net).

ResultsCase historiesDogsOf the 14 dogs with CDV infection, only two were older than oneyear, four were eight to 12 months old and e ight were six monthsold or younger (Table 1). All had high-risk origins or circumstances

such as exposure in an animal shelter, importation, unknown ori-gins or contact with dogs with suspected CDV infection. The onlyanimal with a complete and documented vaccination history was amale seven-year-old American Staffordshire terrier (3148-03) ownedby an Austrian animal shelter. This dog had not been vaccinatedfor 2.5 years when it was presented at the end of April 2003 withfever, diarrhoea, purulent conjunctivitis, bronchopneumonia, gen-eral muscle atrophy, particularly of the masticatory muscles, andmyoclonus of the pelvic limbs. Because of the progression of CNSsigns, the dog was euthanased. All other dogs were either too youngfor (complete) vaccination, not vaccinated or had an unknown vac-cination history.

Clinically, 13 of the 14 dogs showed respiratory signs, 11 hadgastroenteritis, eight had hyperkeratosis and 10 had CNS s igns.Eleven dogs were euthanased or died, while the three survivors

Lednicky and others (2004) described CDV infections in raccoons(Procyon lotor) that differ in virulence and growth characteristics depend-ing on the year of isolation. McCarthy and others (2007) have identi-fied, in particular, two amino acid residues in the H protein associatedwith host specificity. Substitutions in these two positions (549 and 530)are predominantly related to host switches to non-dog hosts. These aresubstitutions of glycine (G) or glutamic acid (E) at position 530 in caninesamples to aspartic acid (D), asparagine (N) or arginine (R) in non-dog

hosts, and substitutions of tyrosine (Y) at position 549 in canine sam-ples to histidine (H) in other hosts.In order to obtain more information about the strains and lineages

of CDV circulating in Austria in both dogs and wildlife, F and H genesequences of selected CDV-positive field samples from the years 2002to 2007 were amplified using specifically designed PCRs and subse-quently submitted for phylogenetic analysis. Amino acid substitutionsrelated to possible cross-species transmission, in particular at positions549 and 530, were analysed.

Materials and methodsBlood samples from 14 dogs, a stone marten (Martes foina) and a badger(Meles meles) were investigated (Table 1).

Primer design and RT-PCR assaysUsing primer design software (Scientific & Educational Software,v4.10), seven primer pairs amplifying a total of more than 3.5 kb ofthe F and H protein genes (Table 2) were designed. RT-PCR assayswere run in a volume of 20 l, made up of 18.4 l reaction mix-ture (OneStep RT-PCR Kit; Qiagen) and 1.6 l template. The cyclerscheme consisted of two pre-PCR steps at 50C for 30 minutes and95C for 15 minutes, followed by 40 cycles at 94C for 30 seconds,55C and 72C for one minute each, and a final extension of 72Cfor 10 minutes.

Sequence and phylogenetic analysesAmplified DNA was extracted using a commercially available kit(NucleoSpin Extract; Machery-Nagel) following the manufacturersinstructions, and served as a template for PCR sequencing. The latter

TABLE 1: Cases of canine distemper virus investigated in 14 dogs, a stone marten (Martes foina) and a badger (Meles meles) in Austriaduring 2002 to 2007

Animal species, sample ID Age Origin or circumstances Vaccination history Clinical signs Outcome

Dog, 4520-02 6 m Imported from Portugal, animal shelter Unknown R+GI EuthanasedDog, 5102-02 6 m No vaccination R+CNS+H DiedDog, 5253-02 1 y Acquired seven weeks before, origin unknown Unknown R+GI+CNS+H EuthanasedDog, 5332-02 1 y No vaccination R+CNS+GI+H EuthanasedDog, 5417-02 4 m Imported, origin unknown Single dose R+GI+H Released to home careDog, 5428-02 1.5 y No vaccination R+GI+CNS EuthanasedDog, 5470-02 1 y Animal shelter Unknown R+CNS+H EuthanasedDog, 5527-02 2 m Imported from Spain, contact animal in Spain died No vaccination R Released to home careDog, 5825-02 5 m In contact with a CDV-positive dog No vaccination R+GI+CNS+H Euthanased

Dog, 3148-03 7 y Animal shelter 2.5 years before R+GI+CNS EuthanasedDog, 4088-03 5 m Imported from Spain, in Austria for four months Single dose R+GI+CNS+H EuthanasedDog, 2779-04 8 m Stray dog found four weeks before onset of clinical signs Single dose GI+CNS Released to home careDog, 2727-07 2 m Imported from Hungary, all littermates died Single dose R+GI+ CNS EuthanasedDog, 2730-07 2 m Three out of a litter of nine alive, dam diseased No vaccination R+GI+H EuthanasedBadger, 3156-06 Adult NA Unknown Found deadStone marten, 2390-07 Adult Abnormal behaviour NA CNS? Killed

CDV Canine distemper virus, GI Gastrointestinal, H Hyperkeratosis, m Months, NA Not applicable, R Respiratory, y Years

TABLE 2: Oligonucleotides used for the amplification of F and H genes (annealing temperature 55C) of selected canine distempervirus-positive samples from Austria during 2002 to 2007

Primer Forward primer sequence Reverse primer sequence Size (bp)Position on strain Onderstepoort(GenBank accession number AF378705) Gene

CDV 1 5-AGCAAGCCAGCCACAGATCG-3 5-TGCACCTGCAAGCACCACTC-3 497 5137-5633 F gene

CDV 2 5-CTCTAATGACCAAGAATGTG-3 5-GCATAACTCAGTGCTTGAAT-3 436 5545-5980 F geneCDV 3 5-TGTGTATTCGTCTCAGAATC-3 5-ATTCTGCTTGAGTGTCTGTT-3 585 6270-6854 F geneCDV 4 5-ACGCTACCAACAGACACTCA-3 5-GAGCGGTGTCAAGACATCTA-3 537 6827-7363 F and H geneCDV 5 5-ATCACCAAGTCATAGATGTC-3 5-ATAGTTGGTTGTCTGGAGTA-3 560 7332-7891 H geneCDV 6 5-TGCCATTACTCCAGACAACC-3 5-TCCATACCGTCTCCATTCAG-3 526 7866-8391 H geneCDV 7 5-CGGTCCGGTTATACTGAATG-3 5-CATGCCTAAGGCCAATTGAG-3 582 8359-8940 H gene

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were released for home care where follow-up investigations werenot possible.

WildlifeBoth the badger and stone marten showed pathological alterations

that were not specifically related to CDV infection, including signsof gastritis.

Sequence analysisBy sequence analysis, the lowest homologies of Austrian samples werefound with the USA-1 cluster, which comprises different old vaccinestrains. Compared with the typical representative, strain Onderstepoort,identities varied from 91 to 94 per cent in segment 1 of the F gene, from93 to 95 per cent in segment 2 of the F gene and from 92 to 93 percent in the H gene. The sequences obtained from dog samples in 2007showed higher levels of identity to strain Onderstepoort than the onesobtained in 2002. Austrian CDV sequences showed homologies to eachother of between 94 and 100 per cent in the F gene, with a higher diver-sity in segment 2 (94 to 100 per cent) than in segment 1 (97 to 100 percent), and of 94 to 100 per cent in the H gene. The wildlife sequences of2006 and 2007 were 98 to 99 per cent identical to each other and morehomologous to the canine samples of 2007 (96 to 97 per cent) than tothose obtained in 2002 (93 to 96 per cent). One sequence (3148-03)showed identities of 96 to 97 per cent in the F gene with PDV type 2(PDV-2) and of 99 per cent in the H gene with several Hungarian Arcticsequences detected in 2005 to 2006.

Phylogenetic analysisBy phylogenetic analysis, the samples showed similar cluster-ing in the three genomic regions investigated, with one exception,3148-03, which groups in the European lineage in segment 1 (posi-tion 5200-5980) of the F gene, but in the Arctic group in segment 2(position 6284-6923) of the F gene (data not shown) and in the H gene(Fig 2). All Austrian CDV sequences detected in 2002 formed a very

closely related group with one 2003 sequence, all well embedded inthe European lineage. The Austrian Arctic sequence showed the high-est identity with Hungarian isolates. The two canine sequences from2007 (2727-07 and 2730-07) occupy a clearly separate position in theEuropean wildlife group/Asia-1 group together with AF178038 (redpanda, China), AF178039 (giant panda, China) and AY964114 (dog,USA). The two Austrian wildlife sequences (3156-06 and 2390-07),which cluster with sequences from a mink (accession numberZ47759, Denmark 1986) and a dog (DQ889189, Hungary 2005), aremore closely related to the latter than to any other Austrian sequence.

Analysis of amino acid positions 530and 549 of the H proteinThe two wildlife amino acid sequences (2006 and 2007) have a sub-

stitution of 549H; all Austrian canine samples (2002, 2003, 2005and 2007) have Y at this position, as described for the majority ofcanine samples worldwide. The wildlife sequences and one caninesample of 2007 (2730-07) have substitutions of 530G/E to 530D, andthe Austrian Arctic sequence (3148-03) has a substitution at the sameposition to 530N (Fig 1).

DiscussionIn the years 2002 to 2007, CDV-positive blood samples from 14 dogs,a badger and a stone marten were available for investigation. Twelve of14 dogs were one year old or younger and, with one exception, eithernot or incompletely vaccinated. The only animal with a complete

and documented vaccination history, but with the last booster admin-istered 2.5 years earlier, was a seven-year-old shelter dog (3148-03). Ina high-risk environment like an animal shelter, recommended boosterintervals of three years have to be considered critically (Schoder andothers 2006). A high percentage of the randomly chosen canine casesshowed CNS involvement (71 per cent), explaining the high fatality(of at least 78 per cent, not including the outcome of dogs released tohome care).

CDV sequences detected in Austria showed homologies of 93to 100 per cent to each other and clustered in different previouslydescribed lineages (Haas and others 1997, Martella and others 2006,Demeter and others 2007). This grouping was similar in all threegenomic regions investigated; only dog 3148-03 clustered in theEuropean lineage in the 5-end of the F gene, but in the Arctic groupin the 3-end of the F gene and in the H gene. This might reflect a pos-sible recombination event.

Circulating CDV strains were largely similar, both in neighbour-ing geographic regions and in the year of detection. All Austrian CDVH gene sequences detected in 2002 formed a group well embeddedin the European lineage, closely related to sequences from Italy in2002 and 2003 and Hungary in 2004, and also to sequences detectedmore than a decade earlier in Germany and Denmark. One sequenceof this cluster detected in 2004 (2779-04) was nearly identical to aHungarian isolate from the same year. The Austrian Arctic sequenceof 2003 showed the highest identities to Hungarian isolates of 2005to 2006 and might have been the source of the Hungarian outbreak.Interestingly, both the Hungarian and the Austrian Arctic sequencesoriginated from dogs kept in animal shelters.

The two sequences from 2007 (2727-07 and 2730-07) occupy a

clearly separate position together with isolates from China (panda,before 2000) and the USA (dog, 2004), which have been assigned toboth the European wildlife and the Asia-1 cluster by different authors(McCarthy and others 2007, An and others 2008, Han and others2008).

The wildlife CDV sequences of 2006 and 2007 were most closelyrelated to a sequence from a Hungarian dog detected in 2005 (acces-sion number DQ889189) and to a mink sequence from Denmark in1986 (accession number Z47759). Although the badger and stonemarten were found nearly a year apart, theMorbillivirus detected wasvery similar in both cases, but clearly distinct from those detectedin dogs. Nevertheless, compared with other Austrian sequences, thewildlife sequences of 2006 and 2007 showed the highest identities (95to 97 per cent) with the canine samples from 2007.

Amino acid sequences for both wildlife samples showed substitu-tions in the two residues of the H protein predominantly related tohost switches and possible changes in virulence (530G/E to 530D and549Y to 549H) (McCarthy and others 2007, Sekulin and others 2011).As expected in CDV sequences, all Austrian canine samples had tyro-sine (Y) at position 549. One sample from 2007 (2730-07) occupied

FIG 1: H protein amino acid alignment of selected Austrian canine distemper virus samples and phocine distemper virus type 2

(PDV-2): GQ214369/2390-07/martesfoina/AUT/2007, GQ214473/3156-06/melesmeles/AUT/2006, GQ214371/2730-07/dog/AUT/2007,

GQ214384/5825-02/dog/AUT/2002, GQ214379/5332-02/dog/2002, GQ214383/5527-02/dog/2002, GQ214380/5417-02/dog/AUT/2002,

GQ214376/4520-02/dog/2002, GQ214378/5253-02/dog/AUT/2002, GQ214373/3148-03/AUT/2003. Positions 530 and 549 are framed.

Reference strain: AY526496, raccoon, USA, 1991

AY526496_rac 1351

X84998_PDV2. 1371GQ214373_dog 1351

GQ214378_dog 1351GQ214376_dog 1351GQ214380_dog 1351GQ214383_dog 64GQ214379_dog 71GQ214384_dog 71GQ214371_dog 57GQ214374_bad 1351GQ214369_mar 1351 . . . . . . . . . . ... . . . . . . . . . . .. . . . . . . . . . . . .. . . . . .. . . . . . .. . . . .. . . . . . . . . .... . . .. . ... . . . . . ... ... . . . . . .. . . . . . . . . ..N DG

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. . . . . .. . . . . . . . . ..N DG. . . . . . . . . . ... . . . . . .. . . . . .. . . . . . . . . . . . .. . . F. .. . . . . . .. . . . .. . . . . . . . . .... . . .. . ... . . . . . ... .K .F .Y . . .. . . . . . . . . ..N D

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USA-2AY49869

2/racc

oon/U

SA/20

01

AF178038/

lesserpand

a/CHN

AY964114/d

og/USA/2004

X84

999/fe

rret/G

ER/198

9

AY6494

46/ra

ccoon/US

A/2001Z

541

56/c

hin

ese

-leo

pard

/US

A/199

AY443350/ta

ccoon/U

SA/2

000A

F164967/d

og

/stra

inA

75-1

5/U

SA

Z47764/ja

velina/U

SA/1

989

Z477

65/raccoon/U

SA/1

989

Z47762/dog/

USA/1989

AY526496/raccoon/U

SA/2001

Z54166/blackpanther/USA/1992

Z47763/blackpanther/USA/1991

AY465925

/racco

on/US

A/2000

AF178039/gia

nt

pand

a/CHN

DQ889189/do

g/HUN

/2005

Z477

59/m

ink/DEN

/1986

EU325729/raccoondog/CHN/2007

D85755/dog/strainYanaka/1992-94

AB016776

/racco

ondog/JPN

/1996

AB2196

5/dog/

JPN

D85754

/dog/stra

inHama

matsu/J

PN/199

2-94

DQ1917

65/dog/T

PE/2004

EU325731

/mink/CHN

/2007

AY964110/do

g/USA/2004

X8499

8/seal

/Siberia

/1987

Z47760

/dog/G

reenla

nd/198

8

AF1724

11/dog/

CHN

EF44505

2/fox/CH

N/2005

DQ22608

7/dog/ITA

/2004

AY964108/do

g/USA/2004

AY964112/dog/USA/2004DQ226088/dog/ITA/2005

GQ214371/2730

-07/d

og/AUT/2007

GQ214370/2727

-07/d

og/AUT/2007

GQ21

4369

/239

0-07

/marte

s

foin

a/AU

T/2007

GQ214374

/3156-06/m

eles

meles/AUT

/2006

DQ889186/dog/HUN/2006

DQ889178/dog/HUN/2005

AB040768/dog

/JPN/1

999

AB

040767/d

og

/JPN

/1999

AB25

2718

/do

g/J

PN

AF378705/fo

x/s

train

Onders

tep

oort/R

SA/1

948

EF418782/d

og/s

train

Lederle/U

SA/1

952

AF259552

/do

g/stra

in

Sn

yderHill/U

SA

/1956

AY

542

312

/raccoo

n/US

A/1998

AY445077/raccoon/USA/1998

AY466011/raccoon/USA/1998

DQ889182/dog/HUN/2005DQ889183/dog/HUN/2005DQ889179/dog/HUN/2005DQ889181/dog/HUN/2005DQ889180/dog/HUN/2005DQ889184/dog/HUN/2006DQ889185/dog/HUN/2006

GQ214373/3148-03/dog/AUT/2003

Arctic-like

Europe

Asia-2

USA-1

0.01

Asia-1

European

wildlife

AY093674/dog/TUR

EU098103/dog/BRA

EU098105/dog/BRA

AF478550/dog/DEN/1991

AF478547/dog/DEN/1991

AF478548/dog/DEN/1991Z47761/dog/DEN/1991Z77671/dog/GER/1994-95

Z77673/dog/GER/1994-95

DQ494317/do

g/ITA/2003

DQ88917

7/dog/HU

N/2004

DQ49

4318/d

og/ITA

/2002

DQ494319/do

g/ITA

/2003

AY386315/d

og/GER/1989

GQ214383/5527

-02/d

og/AUT/2002

GQ214375/4088

-03/

dog/AUT/2003

GQ214

372/27

79-07/

dog/AU

T/2004

GQ214380/5417-02/dog/AUT/2002GQ214379/5332-02/dog/AUT/2002GQ214382/5470-02/dog/AUT/2002GQ214384/5825-02/dog/AUT/2002GQ214376/4520-02/dog/AUT/2002

GQ214377/5101-02/dog/AUT/2002GQ214381/5428-02/dog/AUT/2002

GQ214378/5253-02/dog/AUT/2002

FIG 2: Evolutionary relationship of 83 taxa

(800 bp of the H gene of canine distemper

virus, position 7079-7878). For each GenBank

sequence, the following information is given:

the GenBank accession number and, if indicated,the strain, host, country of origin and year of

detection. Austrian sequences are given in bold

(accession number/specimen voucher/species/

year of detection). The evolutionary history was

inferred using the neighbour-joining method.

The optimal tree with the sum of branch length

= 0.66610640 is shown. The tree is drawn to

scale, with branch lengths in the same units

as those of the evolutionary distances used to

infer the phylogenetic tree. The evolutionarydistances were computed using the maximum

composite likelihood method and are in the units

of the number of base substitutions per site.

Phylogenetic analyses were conducted in MEGA4

(Saitou and Nei 1987, Tamura and others 2004,

2007)



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an intermediate position between wildlife and canine sequences. Inthis sample, there was a substitution at position 530, where asparticacid (D) replaced glycine (G) or glutamic acid (E). This substitutionwas also present in both wildlife samples from 2006 and 2007. Thesefindings might be explained by a previous interspecies transmissionbetween wildlife and dogs followed by viral evolution. The AustrianArctic sequence shows asparagine (N) at position 530, as do severalother Arctic-like sequences.

In conclusion, as described for other countries, different distinctlineages of CDV sequences were present in Austria in the years 2002to 2007, with a strong geographic and temporal clustering. Fourlineages were found in Austria: the canine samples of 2002 to 2005were assignable to the European lineage, one canine sample from2003 belonged to the Arctic lineage, the European wildlife lineagewas represented by two wildlife samples from 2006 and 2007, and afourth cluster was formed by two canine samples from 2007. Aminoacid substitutions related to host specificity were present in the twoAustrian wildlife samples at both positions of the H protein (530 and549) known to be predominantly related to host switching. Of thecanine samples analysed, both the Arctic sample (3148-03) and onesample from 2007 showed substitutions at position 530 but not atposition 549.

All sequences have been deposited at GenBank (www.ncbi.nlm.nih.gov), with the accession numbers: GQ214337-GQ214352(F gene, position 5200-5980 of strain Onderstepoort), GQ214353-GQ214355 and GQ214357-GQ214368 (F gene, position 6284-6923)and GQ214369-GQ214384 (H gene, position 7079-7878 or position7079-8902).

AcknowledgementsThe authors thank R. Skerlak for medical care of the study animals.

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2011 168: 377 originally published online April 6,Veterinary Record

V. Benetka, M. Leschnik, N. Affenzeller, et al.samples from dogs and wild carnivoresdistemper virus strains from clinicalPhylogenetic analysis of Austrian canine

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Phylogenetic Analysis of Austrian Canine Distemper Virus From Clinical Samples From Dogs and Wild...

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