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1.  Consensus proposals for classification of the family Hepeviridae 
The Journal of General Virology  2014;95(Pt 10):2223-2232.
The family Hepeviridae consists of positive-stranded RNA viruses that infect a wide range of mammalian species, as well as chickens and trout. A subset of these viruses infects humans and can cause a self-limiting acute hepatitis that may become chronic in immunosuppressed individuals. Current published descriptions of the taxonomical divisions within the family Hepeviridae are contradictory in relation to the assignment of species and genotypes. Through analysis of existing sequence information, we propose a taxonomic scheme in which the family is divided into the genera Orthohepevirus (all mammalian and avian hepatitis E virus (HEV) isolates) and Piscihepevirus (cutthroat trout virus). Species within the genus Orthohepevirus are designated Orthohepevirus A (isolates from human, pig, wild boar, deer, mongoose, rabbit and camel), Orthohepevirus B (isolates from chicken), Orthohepevirus C (isolates from rat, greater bandicoot, Asian musk shrew, ferret and mink) and Orthohepevirus D (isolates from bat). Proposals are also made for the designation of genotypes within the human and rat HEVs. This hierarchical system is congruent with hepevirus phylogeny, and the three classification levels (genus, species and genotype) are consistent with, and reflect discontinuities in the ranges of pairwise distances between amino acid sequences. Adoption of this system would include the avoidance of host names in taxonomic identifiers and provide a logical framework for the assignment of novel variants.
doi:10.1099/vir.0.068429-0
PMCID: PMC4165930  PMID: 24989172
2.  Schmallenberg Virus Antibodies in Adult Cows and Maternal Antibodies in Calves 
Emerging Infectious Diseases  2014;20(5):901-902.
doi:10.3201/eid2005.130763
PMCID: PMC4012791  PMID: 24751164
Schmallenberg virus; antibodies; cows; calves; novel orthobunyavirus; biting midges; Culicoides spp.; immunity; ruminants; vaccine; vaccination; viruses; the Netherlands; ruminant livestock; maternal antibodies; passive immunity
3.  Hepatitis E Virus in Pork Liver Sausage, France 
Emerging Infectious Diseases  2013;19(2):264-266.
We investigated viability of hepatitis E virus (HEV) identified in contaminated pork liver sausages obtained from France. HEV replication was demonstrated in 1 of 4 samples by using a 3-dimensional cell culture system. The risk for human infection with HEV by consumption of these sausages should be considered to be high.
doi:10.3201/eid1902.121255
PMCID: PMC3563277  PMID: 23347828
Hepatitis E; hepatitis E virus; foodborne disease; pork; food chain; 3D cell culture; viruses; France; HEV; sausage; pork liver; pig liver; contamination
4.  Development and application of a SYBR green RT-PCR for first line screening and quantification of porcine sapovirus infection 
Background
Sapoviruses are single stranded positive sense RNA viruses belonging to the family Caliciviridae. The virus is detected in different species including the human and the porcine species as an enteric pathogen causing asymptomatic to symptomatic enteritis. In this study, we report the development of a rapid real time qRT-PCR based on SYBR Green chemistry for the diagnosis of porcine sapovirus infection in swine.
Results
The method allows the detection of porcine sapoviruses and the quantification of the genomic copies present in stool samples. During its development, the diagnostic tool showed good correlation compared with the gold standard conventional RT-PCR and was ten-fold more sensitive. When the method was applied to field samples, porcine noroviruses from genogroup 2 genotype 11b were also detected. The method was also applied to swine samples from the Netherlands that were positive for PoSaV infection. Phylogenetic results obtained from the samples showed that PoSaV sequences were genetically related to the currently described genogroup III, to the proposed genogroup VII and also to the MI-QW19 sequence (close to the human SaV sequences).
Conclusions
A rapid, sensitive, and reliable diagnosis method was developed for porcine sapovirus diagnosis. It correlated with the gold standard conventional RT-PCR. Specificity was good apart for genogroup 2 genotype 11b porcine noroviruses. As a first line screening diagnosis method, it allows a quicker and easier decision on doubtful samples.
doi:10.1186/1746-6148-8-193
PMCID: PMC3528410  PMID: 23072668
Sapovirus; Diagnosis; Real time PCR; Porcine; SYBR green; Phylogeny
5.  Seroprevalence of Schmallenberg Virus Antibodies among Dairy Cattle, the Netherlands, Winter 2011–2012 
Emerging Infectious Diseases  2012;18(7):1065-1071.
Seroprevalence was highest in the eastern part of the country, bordering Germany, where the virus was first identified.
Infections with Schmallenberg virus (SBV) are associated with congenital malformations in ruminants. Because reporting of suspected cases only could underestimate the true rate of infection, we conducted a seroprevalence study in the Netherlands to detect past exposure to SBV among dairy cattle. A total of 1,123 serum samples collected from cattle during November 2011–January 2012 were tested for antibodies against SBV by using a virus neutralization test; seroprevalence was 72.5%. Seroprevalence was significantly higher in the central-eastern part of the Netherlands than in the northern and southern regions (p<0.001). In addition, high (70%–100%) within-herd seroprevalence was observed in 2 SBV-infected dairy herds and 2 SBV-infected sheep herds. No significant differences were found in age-specific prevalence of antibodies against SBV, which is an indication that SBV is newly arrived in the country.
doi:10.3201/eid1807.120323
PMCID: PMC3376820  PMID: 22709656
Orthobunyavirus; Schmallenberg virus; seroprevalence; within-herd prevalence; dairy cattle; the Netherlands; viruses; Suggested citation for this article: Elbers ARW; Loeffen WLA; Quak S; de Boer-Luijtze E; van der Spek AN; Bouwstra R; et al. Seroprevalence of Schmallenberg virus antibodies among dairy cattle; the Netherlands; winter 2011–2012. Emerg Infect Dis [serial on the internet]. 2012 Jul [date cited]. http://dx.doi.org/10.3201/eid1807.120323
6.  First Isolation of Hepatitis E Virus Genotype 4 in Europe through Swine Surveillance in the Netherlands and Belgium 
PLoS ONE  2011;6(8):e22673.
Hepatitis E virus (HEV) genotypes 3 and 4 are a cause of human hepatitis and swine are considered the main reservoir. To study the HEV prevalence and characterize circulating HEV strains, fecal samples from swine in the Netherlands and Belgium were tested by RT-PCR. HEV prevalence in swine was 7–15%. The Dutch strains were characterized as genotype 3, subgroups 3a, 3c and 3f, closely related to sequences found in humans and swine earlier. The HEV strains found in Belgium belonged to genotypes 3f and 4b. The HEV genotype 4 strain was the first ever reported in swine in Europe and an experimental infection in pigs was performed to isolate the virus. The genotype 4 strain readily infected piglets and caused fever and virus shedding. Since HEV4 infections have been reported to run a more severe clinical course in humans this observation may have public health implications.
doi:10.1371/journal.pone.0022673
PMCID: PMC3148228  PMID: 21829641
7.  Public Health Threat of New, Reemerging, and Neglected Zoonoses in the Industrialized World 
Improving our capacity to respond to these pathogens is essential.
Microbiologic infections acquired from animals, known as zoonoses, pose a risk to public health. An estimated 60% of emerging human pathogens are zoonotic. Of these pathogens, >71% have wildlife origins. These pathogens can switch hosts by acquiring new genetic combinations that have altered pathogenic potential or by changes in behavior or socioeconomic, environmental, or ecologic characteristics of the hosts. We discuss causal factors that influence the dynamics associated with emergence or reemergence of zoonoses, particularly in the industrialized world, and highlight selected examples to provide a comprehensive view of their range and diversity.
doi:10.3201/eid1601.081467
PMCID: PMC2874344  PMID: 20031035
Zoonoses; bacteria; viruses; parasites; infectious diseases; arthropod-borne disease; new zoonoses; emerging diseases; reemerging infections; synopsis
8.  Self-Assembly of the Recombinant Capsid Protein of a Swine Norovirus into Virus-Like Particles and Evaluation of Monoclonal Antibodies Cross-Reactive with a Human Strain from Genogroup II▿  
Journal of Clinical Microbiology  2008;46(12):3971-3979.
Noroviruses (NoVs) are responsible for the majority of gastroenteritis outbreaks in humans. Recently, NoV strains which are genetically closely related to human genogroup II (GII) NoVs have been detected in fecal specimens from swine. These findings have raised concern about the possible role of pigs as reservoirs for NoVs that could infect humans. To better understand the epidemiology of swine NoVs in both the swine and the human populations, rapid immunoassays are needed. In this study, baculovirus recombinants were generated to express the capsid gene of a swine NoV GII genotype 11 (GII.11) strain which self-assembled into virus-like particles (VLPs). Subsequently, the purified VLPs were used to evoke monoclonal antibodies (MAbs) in mice. A panel of eight promising MAbs was obtained and evaluated for their ability to bind to heterologous VLPs, denaturated antigens, and truncated capsid proteins. The MAbs could be classified into two groups: two MAbs that recognized linear epitopes located at the amino-terminal half (shell domain) of the swine NoV GII.11 VLPs and that cross-reacted with human GII.4 NoV VLPs. The other six MAbs bound to conformational epitopes and did not cross-react with the human GII.4 VLPs. To our knowledge, this is the first report on the characterization of MAbs against swine NoVs. The swine NoV VLPs and the MAbs described here may be further used for the design of diagnostic reagents that could help increase our knowledge of the prevalence of NoV infections in pigs and the possible role of pigs as reservoirs for NoVs.
doi:10.1128/JCM.01204-08
PMCID: PMC2593275  PMID: 18842943
9.  European Bat Lyssaviruses, the Netherlands 
Emerging Infectious Diseases  2005;11(12):1854-1859.
Genotype 5 lyssaviruses are endemic in the Netherlands, and can cause fatal infections in humans.
To study European bat lyssavirus (EBLV) in bat reservoirs in the Netherlands, native bats have been tested for rabies since 1984. For all collected bats, data including species, age, sex, and date and location found were recorded. A total of 1,219 serotine bats, Eptesicus serotinus, were tested, and 251 (21%) were positive for lyssavirus antigen. Five (4%) of 129 specimens from the pond bat, Myotis dasycneme, were positive. Recently detected EBLV RNA segments encoding the nucleoprotein were sequenced and analyzed phylogenetically (45 specimens). All recent serotine bat specimens clustered with genotype 5 (EBLV1) sequences, and homologies within subgenotypes EBLV1a and EBLV1b were 99.0%–100% and 99.2%–100%, respectively. Our findings indicate that EBLVs of genotype 5 are endemic in the serotine bat in the Netherlands. Since EBLVs can cause fatal infections in humans, all serotine and pond bats involved in contact incidents should be tested to determine whether the victim was exposed to EBLVs.
doi:10.3201/eid1112.041200
PMCID: PMC3367619  PMID: 16485470
EBLV; lyssavirus; the Netherlands; bat; Eptesicus serotinus; Myotis dasycneme; Europe; research
10.  Phylogeography, Population Dynamics, and Molecular Evolution of European Bat Lyssaviruses 
Journal of Virology  2005;79(16):10487-10497.
European bat lyssaviruses types 1 and 2 (EBLV-1 and EBLV-2) are widespread in Europe, although little is known of their evolutionary history. We undertook a comprehensive sequence analysis to infer the selection pressures, rates of nucleotide substitution, age of genetic diversity, geographical origin, and population growth rates of EBLV-1. Our study encompassed data from 12 countries collected over a time span of 35 years and focused on the glycoprotein (G) and nucleoprotein (N) genes. We show that although the two subtypes of EBLV-1—EBLV-1a and EBLV-1b—have both grown at a low exponential rate since their introduction into Europe, they have differing population structures and dispersal patterns. Furthermore, there were strong constraints against amino acid change in both EBLV-1 and EBLV-2, as reflected in a low ratio of nonsynonymous to synonymous substitutions per site, particularly in EBLV-1b. Our inferred rate of nucleotide substitution in EBLV-1, approximately 5 × 10−5 substitutions per site per year, was also one of the lowest recorded for RNA viruses and implied that the current genetic diversity in the virus arose 500 to 750 years ago. We propose that the slow evolution of EBLVs reflects their distinctive epidemiology in bats, where they occupy a relatively stable fitness peak.
doi:10.1128/JVI.79.16.10487-10497.2005
PMCID: PMC1182613  PMID: 16051841

Results 1-10 (10)