Gábor Reuter, Mateja Polj ak-Prijatelj, Ilaria Di Bartolo, …jcm.asm.org/content/early/2009/11/25/JCM.01279-09.full.pdf4 Maria Ruggeri, 3 Tuija Kantala, 4 Leena Maunula, 4 István

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Incidence, diversity and molecular epidemiology of sapoviruses in swine across Europe 1

Running title: Porcine sapoviruses in Europe 2

Gábor Reuter,1* Janet Zimšek-Mijovski,2 Mateja Poljšak-Prijatelj,2 Ilaria Di Bartolo,3 Franco 3

Maria Ruggeri,3 Tuija Kantala,4 Leena Maunula,4 István Kiss,5 Sándor Kecskeméti,5 Nabil 4

Halaihel,6 Javier Buesa,7 Christina Johnsen,8 Charlotte K Hjulsager,9 Lars E Larsen,9 Marion 5

Koopmans,10 Blenda Böttiger8 6

7

ÁNTSZ Regional Institute of State Public Health Service, Pécs, Hungary1 8

University of Ljubljana, Ljubljana, Slovenia2 9

Istituto Superiore di Sanita, Rome, Italy3 10

University of Helsinki, Helsinki, Finland4 11

Central Agricultural Office, Veterinary Diagnostic Directorate, Debrecen, Hungary5 12

University of Zaragoza, Zaragoza, Spain6 13

University of Valencia, Valencia, Spain7 14

Statens Serum Institut, Copenhagen, Denmark8 15

Technical University of Denmark, National Veterinary Institute, Copenhagen, Denmark9 16

National Institute for Public Health and the Environment, Bilthoven, The Netherlands10 17

* Address for correspondence: 18

Dr Gábor Reuter 19

Regional Laboratory of Virology, 20

ÁNTSZ Regional Institute of State Public Health Service 21

Szabadság út 7. H-7623 Pécs, Hungary 22

Telephone: +36 06 (72) 514 979 23

Fax: +36 06 (72) 514 949 24

Email: [email protected] 25

Copyright © 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.J. Clin. Microbiol. doi:10.1128/JCM.01279-09 JCM Accepts, published online ahead of print on 25 November 2009

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Abstract 26

Porcine sapovirus is an enteric calicivirus in domestic pigs belonging to the family 27

Caliciviridae. Some porcine sapoviruses are genetically related to human caliciviruses, which 28

has raised public health concerns of animal reservoirs and potential cross-species transmission 29

for sapoviruses. In this comprehensive study we report the incidence, genetic diversity and 30

molecular epidemiology of sapoviruses detected in domestic pigs in six European countries, 31

Denmark, Finland, Hungary, Italy, Slovenia and Spain, between 2004 and 2007. A total of 32

1050 swine fecal samples from 88 pig farms were collected and tested by reverse 33

transcription-polymerase chain reaction for sapoviruses, and positive findings were confirmed 34

by sequencing. Sapoviruses were detected in 80 (7.6%) samples collected on 39 (44.3%) 35

farms, and in every country. The highest prevalence was seen among piglets aged 2-8 weeks 36

and – where tested (Spain and Denmark) - there was no significant difference in the 37

proportion of sapovirus positive findings in healthy animals or animals with diarrhea. Based 38

upon the RNA-polymerase region highly heterogeneous populations of viruses were identified 39

representing 6 different genogroups (III, VI, VII, and VIII) including potential new 40

genogroups (IX and X) with a predominance of GIII (50.6%). Genogroup VIII, found in 5 of 41

the 6 countries, had the highest homology (up to 66% at amino acid level) to human sapovirus 42

strains. Sapoviruses are commonly circulating and endemic agents in swine herds Europe-43

wide. Highly heterogeneous, and potential new genogroups of sapoviruses were found in pigs, 44

however, any “human-like” sapovirus was not detected. 45


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Introduction 46

Caliciviruses (family Caliciviridae) are small, non-enveloped viruses with single-47

stranded, positive-sense genomic RNA which are classified – at present – into five genera: 48

Vesivirus, Lagovirus, Norovirus, Nebovirus and Sapovirus (14, 49

http://talk.ictvonline.org/media/p/1203.aspx). Viruses within one calicivirus genus are 50

phylogenetically related, and have the same genomic organization (5). The sapovirus genome 51

is 7.3 to 7.5 kb in length and contains two main open reading frames (ORFs). Sapoviruses are 52

important enteric pathogens that can cause diarrhea in humans, pigs and mink (5, 6, 16). 53

Based upon phylogenetic clustering of the capsid gene (ORF1) and protein sequences, 54

sapoviruses have been classified into five distinct genogroups (GI to GV) (3). Human 55

sapoviruses belong to GI, GII, GIV and GV, whereas porcine sapovirus belongs to GIII. 56

Recently, new porcine sapovirus genogroups (GVI, GVII, GVIII) were proposed (15, 25, 27). 57

Recombinant sapoviruses - both in human and swine hosts - have also been described (8, 24). 58

Each genogroup can be further divided into genetically diverse genotypes. 59

Porcine sapovirus (historically called porcine enteric calicivirus – PEC) was first 60

identified in the United States by electron microscopy in 1980 (21) and genetically 61

characterized as a sapovirus in 1999 (5, 24). Porcine sapoviruses have been reported in only a 62

few additional countries: in The Netherlands (van der Heide et al., available only in the 63

GenBank database), in South Korea (11), Venezuela (17), Hungary (20) and recently in Italy 64

(16), Brazil (1) and Canada (13). The porcine sapovirus strains Cowden and LL14/02/US, 65

representatives of sapovirus GIII, were detected in diarrheic piglets and have been shown to 66

induce enteric diseases and lesions in experimentally infected pigs (4, 7). 67

A public health concern of potential cross-species transmission and animal reservoirs 68

for sapoviruses has been raised. In this study, we report the incidence, genetic diversity and 69

molecular epidemiology of sapoviruses detected in domestic pigs in six European countries. 70


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Materials and Methods 71

Stool samples. 72

A total of 1050 fecal samples from domestic pigs (Sus scrofa domestica) collected at 73

88 swine farms in six European countries (Denmark, Finland, Hungary, Italy, Slovenia and 74

Spain) from January 2004 to December 2007 were tested for sapoviruses (Table 1). Detailed 75

information of sampling periods, age groups, number of tested farms and animals per country 76

is summarized in Table 1. Farms and sampled animals were randomly selected. Clinically 77

healthy animals, without signs of gastroenteritis, were tested in Finland, Hungary, Italy and 78

Slovenia; however, both healthy and diarrheic animals were tested in Denmark and Spain. 79

Fresh fecal samples were placed into sterile containers and stored frozen at -20ºC until tested. 80

RNA extraction and RT-PCR. 81

The RNA was extracted from fecal suspensions according to the standard RNA 82

extraction methods used by each laboratory. RT-PCR was also performed by the participating 83

laboratories using the generic calicivirus primer pair p290/p289 (10), a degenerated version of 84

these primers p290D/p289D (3) or a specific sapovirus primer pair SR80/JV33 (23), all 85

designed for the RNA-dependent RNA polymerase region (Table 1). PCR-products of primer-86

pair p289/p290, p290D/p289D and SR80/JV33 are 331-nt (95-aa), 331-nt and 320-nt for 87

sapovirus, respectively. 88

Sequence and phylogenetic analysis. 89

PCR-products were sequenced directly in both directions using the PCR primers. 90

Sequences were analyzed using the GeneDoc version 2.6 program (19). A dendrogram was 91

constructed using the UPGMA method by MEGA version 3.1 (12). Maintaining the 92

continuity of the current nomenclature for sapovirus genogrouping, we used the previously 93

proposed taxonomic names (25). 94

Statistics. 95


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To test the differences in proportions of double and single infections in animals with 96

diarrhea, Fischer’s exact test was used, and a single-tailed p-value of < 0.5 was considered a 97

significant difference. The association between the incidence of sapovirus infection and 98

diarrhea was tested by chi square analysis. 99

Nucleotide sequence accession numbers. 100

Sapovirus sequences from pigs were submitted – in batch sets for each country - to 101

GenBank under accession numbers FJ854507-FJ854541 (Denmark); FJ861075-FJ861076 and 102

FJ866501-FJ866503 (Finland); DQ383274, FJ808729-FJ808731 (Hungary); GQ228085-103

GQ228090 (Italy); FJ715777-FJ715805 (Slovenia); FN397821 (Spain). 104

Results 105

A total of 117 (11.1%) samples from 1050 pigs yielded a PCR fragment with the 106

expected size for sapoviruses following RT-PCR and gel-electrophoresis. Two thirds (N=80) 107

of these (68% of gel-positives; 7.6% of samples tested) was confirmed as sapovirus specific 108

by sequence analysis of the PCR-products (Table 1). The low amount of DNA obtained for 109

some samples did not allow performing sequence reactions for all samples. In addition, 110

bacterial and host DNA sequences with expected size (false positive) were also amplified 111

using p289/p290 primers. Double infections were detected in 7 pigs increasing the total 112

number of sapovirus sequences available for study to 87. All these sequences contained the 113

conserved amino acid motif GLPSG of the calicivirus RNA-dependent RNA polymerase. The 114

incidence of the sapovirus-positive samples ranged between 1.6% (Hungary) and 49.1% 115

(Denmark). Samples from 39 (44.3%) of the 88 farms included were sapovirus PCR-positive, 116

ranging from 7% in Spain to 87.5% in Slovenia. 117

The number of the fecal samples positive by age-group and the incidence of sapovirus-118

positive animals by age-groups were summarized in Table 1. The sapovirus incidence ranged 119

between 0% and 66.7% within each age group (Table 1). In Finland, Hungary, Italy and 120


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Slovenia all samples – both sapovirus positive and negative - were obtained from clinically 121

healthy pigs, while in Denmark and Spain samples from both, with and without diarrhea, were 122

tested. In Denmark, sapoviruses were detected in 14/30 (47%) and in 14/27 (52%) samples 123

from animals with and without diarrhea, respectively. The corresponding proportions in Spain 124

were 13/113 (12%) and in 14/108 (13%), respectively. No significant differences in the 125

proportion of sapovirus positive samples among pigs with or without diarrhea were seen. 126

A total of 81 out of 87 sapovirus polymerase sequences could be used for further 127

analysis: 35 (43.2%) sequences from Denmark, 5 (6.2%) from Finland, 7 (8.6%) from 128

Hungary, 5 (6.2%) from Italy, 28 (34.6%) from Slovenia and 1 (1.2%) from Spain. The 129

phylogenetic analysis divided the sapovirus sequences into six different genogroups (Figure 130

1). Most sapoviruses belonged to the genogroup GIII (N=41; 50.6%) but GVI (N=1; 1.2%), 131

GVII (N=11; 13.6%) and GVIII (N=6; 7.4%) were also confirmed. Furthermore, strains 132

belonging to two potentially new sapovirus genogroups, tentatively classified as GIX (N=8; 133

9.9%) and GX (N=14; 17.3%) were also detected (Figure 1). By phylogenetic analysis, two 134

lineages (A and B) of GIII strains were found (Figure 1). All six genogroups were identified 135

in Denmark. Three genogroups, GIII, GVIII and GX were found in at least four of the 136

countries studied, whereas GVI, GVII and GIX were detected only in one or two countries. 137

For comparison of sapovirus strains found in pigs and in humans, we compared 138

nucleotide and amino acid sequence identities of the RNA-polymerase gene between the 139

study strains in pigs and the prototype sapovirus genogroups in humans (Table 2). The highest 140

sequence identities (up to 66% in amino acid) were found between genogroup GVIII and all 141

human sapovirus genogroups. GVIII strains in pigs are related to each other with 63-87% 142

nucleotide and 76-98% amino acid identity, respectively. The lowest sequence identity was 143

seen between genogroup IX and human sapoviruses. Between the two GIII lineages, GIIIA 144

and GIIIB, the nucleotide and amino acid identities were 68-84% and 75-91%, respectively. 145


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The distribution of sapovirus genogroups over time (figure not shown) and by age 146

groups (Figure 2) was further analysed. In the first 2 years of the study, genogroup GIII 147

strains were detected most commonly (86%), but this data was from Hungary and Slovenia 148

only, whereas strains belonging to six genogroups (GIIIA and B, GVI, GVII, GVIII, GIX and 149

GX) were found in 5 countries in the last two years (2006 and 2007). Clustering of genotypes 150

by age showed that the dominant GIII strains were detected primarily in young pigs (78%, 151

49% and 0% of strains from pigs in age groups less than 1 month, 1-3 months and more than 152

3 months, respectively). GVI and GVI genotype strains were identified only in age group 1-3 153

months. 154

Double infections with two sapoviruses from different genogroups were identified in 7 155

animals (Vet4 and Vet4B, GVII/GIX; VET19 and Vet19B, GVIII/GX; Vet21A and Vet21B, 156

GIII/GVII; Vet23 and Vet23B, GIII/GVII; Vet25 and Vet25B, GIX/GIII; Vet31 and Vet31B, 157

GVII/GX, Vet33A and Vet33B, GIX/GX) under age of 12 weeks. The different virus types 158

were detected by the use of different diagnostic primer pairs. Six of the seven animals with a 159

double infection had diarrhoea, which was significantly more than seen in the 21 animals 160

infected with a single sapovirus and where only 8 animals had diarrhoea in Denmark. 161

Furthermore, identical sapovirus sequences were detected within the same herds in 6 animal 162

pairs in Denmark (N=4) and Hungary (N=2). 163

Discussion 164

Only few studies have been done to investigate the molecular epidemiology of 165

sapoviruses in swine, since the discovery of porcine sapovirus in 1980 (21). In this study, we 166

describe a large, integrated analysis related to the incidence, genetic diversity and molecular 167

epidemiology of porcine sapoviruses in six European countries. 168

Based upon our data, approximately 8% of tested animals were infected with 169

sapoviruses and shedding the virus in the feces, and sapoviruses were present in more than 170


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40% of the selected swine farms in Europe, indicating that sapoviruses are circulating as 171

endemic agents in swine herds. However, the variation of sapoviruses incidence in individual 172

pigs, as well as among farms, in the participating countries was considerable (data not shown). 173

This could be a reflection of the different possibilities to obtain samples in each of the 174

participating countries. The overall sapovirus positivity rate in our study is lower than in some 175

other studies showing a prevalence of sapoviruses of at least 62% in 621 pigs from 7 different 176

farms in the USA (26), of 30% among 113 piglets from 34 farms in Brazil (1), and of 23% in 177

53 post-weaning pigs from 5 farms in Korea (28), but considerable ranges have been 178

published as well (25). The primers used and selection of samples tested regarding age and 179

state of health differed also in these studies making direct comparisons between them difficult, 180

but it can be concluded that sapovirus infections among pigs are common events. However, 181

our data confirms that pigs are infected with sapoviruses early in life periods in line with the 182

studies in Venezuela, Brazil, Korea and USA (17, 1, 9, 26). As also described in the papers 183

from Venezuela, Brazil and Belgium (18) we found sapoviruses in nearly equal numbers in 184

pigs with and without diarrhea as well. However, diarrhea was significantly associated with 185

co-infection with different sapovirus strains. This does not necessarily imply that the double 186

infection itself causes diarrhea, but might reflect lower hygienic standard in the pens of these 187

pigs, and other pathogens could therefore also be present. 188

It should be also noted that double infections were detected only with using different 189

primer-sets indicating that there is a diagnostic gap in sapovirus detection. Against this, 190

extremely diverse groups of sapoviruses were detected among swine in this study. Porcine 191

strains clustered into a total of 4 known and 2 - previously unknown - potential new 192

genogroups (GIX and GX). On the other hand, genotype III strains were detected most 193

frequently. Their predominance in a limited time period and in young animals suggests that 194

the pigs were sampled during a GIII epidemic, but coordinated studies across regions or 195


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countries are needed to better understand the dynamics of genogroup(s) changes and the 196

possibilities of drift and international spread of sapoviruses among pigs. 197

Recently, it was shown that in some cases there was no sharp demarcation of host-198

species for some calicivirus infections; some caliciviruses may have zoonotic potential and 199

animals such as domestic pigs may act as a reservoir for norovirus or vesivirus (24, 2, 22). By 200

phylogenetic analysis, a higher genetic diversity was seen among porcine sapoviruses than 201

within the known human sapoviruses indicating a longer co-evolution of sapoviruses in swine. 202

One sapovirus genogroup, GVIII, occupy a special position among sapoviruses. GVIII is 203

genetically more closely related to human sapoviruses (especially GV and GI) than to other 204

sapovirus genogroups in swine (25, 16). A previous study suggested that this strain circulates 205

infrequently and has low levels of virus copies in the feces of pigs and therefore probably 206

originates from a non-porcine host (25). Interestingly, in our study, these unique sapovirus 207

strains were found in 5 of the 6 countries constituting 7.4% of the positive sapovirus samples. 208

This indicates a Europe-wide circulation and a relatively frequent incidence of this strain in 209

pigs. Until now, GVIII or GVIII-like strains have not been detected in humans. On the other 210

hand, among the known sapoviruses in swine, the GVIII strains have the closest evolutionary 211

cross-point between human and porcine sapoviruses, as shown by sequence and phylogenetic 212

analysis. 213

Based on this comprehensive study no evidence for zoonotic sapovirus could be found 214

in pigs, but a surprisingly diverse profile of sapoviruses was observed in European swine 215

herds. While the clinical relevance remains to be determined, it reflects how little we know 216

about the viral pathogens present in these food animals. Knowledge about their potential for 217

spread, including contribution to recombination events as was described for sapoviruses 218

belonging to different genogroups is important to understand the role of swine as a potential 219

reservoir. 220


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Acknowledgements 221

This work was supported by grants from the Hungarian Scientific Research Fund 222

(OTKA, F048433) and the project „Enteric Virus Emergence, New Tools” (EVENT, SP22-223

CT-2004-502571) funded by the European Union. 224

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Figure legends 302

Figure 1. Phylogenetic analysis of sapoviruses including porcine sapoviruses detected in this 303

study in Europe based upon the 286-nt long RNA-dependent RNA polymerase region of 304

caliciviruses. A dendrogram was constructed using the UPGMA method by MEGA (version 305

3.1). The country-origin of the strains is indicated by colour coding (Denmark: red, Finland: 306

light-blue, Hungary: blue, Italy: purple, Slovenia: green, Spain: yellow). To maintain the 307

continuity of the current nomenclature for sapovirus genogrouping we used the previously 308

proposed taxonomic names as a basis (Wang et al., 2005, Martella et al., 2008). Sapoviruses 309

in GI, GII, GIV and GV are detected in humans. Swine-origin sapoviruses belongs to GIII, 310

GVI, GVII, GVIII, GIX and GX. Reference strains were obtained from GenBank: 311

PEC/Cowden/US (AF182760), PECLL14/US (AY425671), PECIVA36/NL (AY615805), 312

Korean6802 (AY289186), London/92/UK (U95645), HUNs11/2000/HUN (AF488717), 313

HUNs12/2000/HUN (AF488718), HUNs17/2000/HUN (AF488720), HUN3739/2008/HUN 314

(FJ844411), Sapporo/82/JP (U77903), Houston/90/US (U95644), Po/SV/Yaracuy/1999/VE 315

(AY633966), Po/SV/Miranda/2000/VE (AY633963), Po/SV/Miranda2/2001/VE (AY633965), 316

PEC-Korean10802 (AY289188), OH-JJ259-00-US (AY826423), NC-QW270-03-US 317

(AY826426), SWECI/VA10/NL (AY615807), OH-MM280-03-US (AY823308), 318

MEX335/1991/MX (AY157869), PAN-1/78/US (AF091736), 43-06-18-p-3-6-ITA 319

(AB221477), SWECIII/VA112/NL (AY615814), OH-LL26/2002/US (AY974195), OH-320

JJ681/2000/US (AY974192), Norwalk (M87661), Mex14917/00/MX (AF435810), 321

Mex340/90/MX (AF435809), cruiseship/00/US (AY157863), Mc10/00/TH (AY237420), 322

C12/00/JP (AY603425), Hou7-1181-90-US (AF435814), Arg39/Arg (AF405715), 323

MEC/1/1999/US (AF338404), FCV (M86379), NB-like (AY082891), K7JP (AB221130), 324

SWECII/VA103/NL (AY615811), MI-QW19-02-US (AY826424). 325

326


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Figure 2. Prevalence of sapovirus genogroups by age group based upon 78 sapovirus strains. 327

Y axis represents the prevalence of genogroups in percentage (%). Count numbers in each bar 328

slice represents sample size for genogroup. Total sample sizes per each age group are at the 329

top of the bars. 330


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Table 1. Incidence of sapoviruses in fecal samples collected from domestic pigs from 88 randomly selected swine farms in six European 1

countries between January 2004 and December 2007. Information of sampling periods, age groups, primers, number of tested and positive farms 2

and animals per country are shown. Samples from healthy animals were collected in Finland, Hungary, Italy, and Slovenia, whereas samples 3

from both healthy and diarrheic animals were collected in Denmark and Spain. 4

5


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Table 2. Nucleotide and amino acid sequence identities (range for all strains sequenced) based 1

upon the RNA-polymerase region between the study strains detected in pigs (rows) and the 2

sapovirus genogroup prototype of human origin (columns). Bold numbers indicate the highest 3

nucleotide and amino acid identities. 4

5

human

swinea

GI

Sapporo/82/JP

(U77903)

GII

London/92/UK

(U95645)

GIV

Hou-7-1181-90-US

(AF435814)

GV

Arg39/Arg

(AF405715)

51-55%

51-55%

52-57%

50-54%

54-58%

50-54%

52-56%

49-52%

GIII A (N=20)b

nt

aa

GIII B (N=21)

nt

aa

50-59%

52-62%

54-59%

50-60%

55-60%

53-57%

53-55%

46-49%

GVI (N=1)

nt

aa

49%

41%

50%

42%

52%

43%

45%

32%

GVII (N=11)

nt

aa

42-46%

36-38%

44-48%

35-41%

44-47%

36-42%

41-45%

33-36%

GVIII (N=6)

nt

aa

50-62%

52-63%

53-64%

53-65%

54-65%

55-66%

54-60%

52-55%

GIX (N=8)

nt

aa

40-42%

33-36%

39-46%

33-37%

40-47%

36-41%

41-44%

29-30%

GX (N=14)

nt

aa

47-52%

37-41%

44-52%

37-43%

44-56%

38-43%

41-48%

30-35%

6

aall study strains (N=81) are included for sequence analysis in each genogroup as separated in 7

Figure 1. For homology comparison, percentage results of the lowest and the highest nt 8

(nucleotide) and aa (amino acid) strain identities are indicated in each box as a percentage 9

range in each genogroup. 10

bnumber of sequences analyzed 11


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Figure 1. 1

PEC/Cow

den/

US

PECLL

14/U

S

Kore

an68

02

Po/S

V/M

iranda/2

000/V

E

Po/S

V/M

iranda2/2

001/V

E

PEC-K

ore

an10802

Po/S

V/Y

ara

cuy/1

999/V

EVet-25B

-DK

Vet-26b-D

KPE

CIV

A36/N

LV

P12-S

LO

P61-S

LO

P6-S

LO

P32-S

LO

P43

-SLO

Vet

-9-D

K

P87

-SLO

4472

-3L2

-ESP

Vet-3

2-DK

P60-S

LO

OH-J

J-25

9-00

-US

NC-QW

270-

03-U

S

Id2/

5/HUN

Id3/5/HUN

PoS3/7/HUNPoS6/7/HUNVet-55-DKVet-21A-DK

Vet-23-DK

338-1-FI

14-351-3-FI

P89-SLOVet-30-DKPoS9/7/HUNPoS10/7/HUNSWECI/VA10/NL

OH-MM280-03-US

Vet-47-DK

P21-SLO

P80-SLO

P81-SLO

P50-SLO

P66-SLO

P10-S

LO

P78

-SLO

P79-S

LO

P48-S

LO

P67-S

LO

P70-S

LO

P65-S

LO

P73-S

LO

P72-S

LO

P77-SLO

SW

ECII/VA10

3/NL

MI-Q

W19

-02-

US

Vet-28-D

K

Vet-19-D

K

P71-S

LO

43-0

6-1

8-p

3-6

-ITA

PoS7/7

/HU

N355-5

-FI

Sw

19-5

/06/IT

Arg

39/A

rgN

B-lik

eM

EC

/1/1

999/U

SH

UN

3739

Housto

n90

Mex1

491

7/0

0/M

X

Sap

poro

/82/

JP

Mc1

0/00/

TH

C12

/00/JP

cruise

ship/00/US

Mex

340/90

/MX

Hou7-

1181

-90-

US

London92

/UK

HUNs11/2000/H

UN

MEX335/1991/MX

HUNs12/2000/HUN

HUNs17/2000/HUN

PAN-1/78/U

S

FCV

Vet-21B

-DK

Vet-23B-D

K

Vet

-37-

DK

Vet-7-D

K

OH-LL26

/2002

/US

Vet-22

-DK

Vet-29-DK

K7JPVet-31-DKVet-4-DKP5-SLOP39-SLO

P49-SLO

OH-JJ681/2000/USVet-57-DKVet-3-DKVet-4B-DK

Vet-33A-DK

Vet-25-DK

Sw35-6/06/IT

Vet-56-DK

Sw17-5/06/IT

Sw36-2/06/IT

Vet-6-DK

Vet-52-D

K

Vet-1

9B-D

K

9-T

EUR-2

4-F

I

Vet-4

0-D

K

Sw

34-2

/06/IT

Vet-1

-DK

Vet-2

-DK

Vet-3

3B

-DK

Vet-3

1B

-DK

Vet-8

-DK

294-0

7-S

LO

SW

EC

III/VA

112/N

LVet-3

8-D

K8-T

EUR-2

3-FI

Norw

alk

0.00.10.20.30.40.50.6 2

GII (genotypes 1-5)

Denmark-DK (35)

Finland-FI (5)

Hungary-HUN (7)

Italy-IT (5)

Slovenia-SLO (28)

Spain-ESP (1)

B (?)

A (?)

GVIII

GV

GI (genotypes 1-3)

GIV (recombinant)

GVII

GVI

GIII

GIX (?)

GX (?)


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Figure 2.

4 15

13

71

11

1

5

3

22

3

101

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

<1 month 1-3 months >3 months

GIIIA GIIIB GVI GVII GVIII GIX GX

N=57 N=14 N=7


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Gábor Reuter, Mateja Polj ak-Prijatelj, Ilaria Di Bartolo, …jcm.asm.org/content/early/2009/11/25/JCM.01279-09.full.pdf4 Maria Ruggeri, 3 Tuija Kantala, 4 Leena Maunula, 4 István

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