Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

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Validation of biomarkers of food intake-critical assessment of candidate biomarkers
Dragsted, Lars O. ; Gao, Qinfeng ; Scalbert, Augustin ; Vergères, Guy ; Kolehmainen, Marjukka ; Manach, Claudine ; Brennan, Lorraine ; Afman, L.A. ; Wishart, David S. ; Lacueva, Cristina Andres ; Garcia-Aloy, Mar ; Verhagen, H. ; Feskens, E.J.M. ; Feskens, E.J.M. ; Praticò, Giulia - \ 2018
Genes & Nutrition 13 (2018). - ISSN 1555-8932
Biomarkers of food intake (BFIs) are a promising tool for limiting misclassification in nutrition research where more subjective dietary assessment instruments are used. They may also be used to assess compliance to dietary guidelines or to a dietary intervention. Biomarkers therefore hold promise for direct and objective measurement of food intake. However, the number of comprehensively validated biomarkers of food intake is limited to just a few. Many new candidate biomarkers emerge from metabolic profiling studies and from advances in food chemistry. Furthermore, candidate food intake biomarkers may also be identified based on extensive literature reviews such as described in the guidelines for Biomarker of Food Intake Reviews (BFIRev). To systematically and critically assess the validity of candidate biomarkers of food intake, it is necessary to outline and streamline an optimal and reproducible validation process. A consensus-based procedure was used to provide and evaluate a set of the most important criteria for systematic validation of BFIs. As a result, a validation procedure was developed including eight criteria, plausibility, dose-response, time-response, robustness, reliability, stability, analytical performance, and inter-laboratory reproducibility. The validation has a dual purpose: (1) to estimate the current level of validation of candidate biomarkers of food intake based on an objective and systematic approach and (2) to pinpoint which additional studies are needed to provide full validation of each candidate biomarker of food intake. This position paper on biomarker of food intake validation outlines the second step of the BFIRev procedure but may also be used as such for validation of new candidate biomarkers identified, e.g., in food metabolomic studies.
Artificial light at night shifts daily activity patterns but not the internal clock in the great tit (Parus major)
Spoelstra, Kamiel ; Verhagen, Irene ; Meijer, Davy ; Visser, Marcel E. - \ 2018
Proceedings of the Royal Society. B: Biological Sciences 285 (2018)1875. - ISSN 0962-8452
Artificial light at night - Circadian period - Circadian phase - Entrainment - Light pollution
Artificial light at night has shown a dramatic increase over the last decades and continues to increase. Light at night can have strong effects on the behaviour and physiology of species, which includes changes in the daily timing of activity; a clear example is the advance in dawn song onset in songbirds by low levels of light at night. Although such effects are often referred to as changes in circadian timing, i.e. changes to the internal clock, two alternative mechanisms are possible. First, light at night can change the timing of clock controlled activity, without any change to the clock itself; e.g. by a change in the phase relation between the circadian clock and expression of activity. Second, changes in daily activity can be a direct response to light (‘masking’), without any involvement of the circadian system. Here, we studied whether the advance in onset of activity by dim light at night in great tits (Parus major) is indeed attributable to a phase shift of the internal clock.We entrained birds to a normal light/dark (LD) cycle with bright light during daytime and darkness at night, and to a comparable (LDim) schedule with dim light at night. The dim light at night strongly advanced the onset of activity of the birds. After at least six days in LD or LDim,we kept birds in constant darkness (DD) by leaving off all lights so birds would revert to their endogenous, circadian system controlled timing of activity.We found that the timing of onset in DD was not dependent on whether the birds were kept at LD or LDim before the measurement. Thus, the advance of activity under light at night is caused by a direct effect of light rather than a phase shift of the internal clock. This demonstrates that birds are capable of changing their daily activity to low levels of light at night directly, without the need to alter their internal clock.
What drives farmers to increase soil organic matter? Insights from the Netherlands
Hijbeek, R. ; Pronk, A.A. ; Ittersum, M.K. van; Berge, H.F.M. ten; Bijttebier, J. ; Verhagen, A. - \ 2018
Soil Use and Management 34 (2018)1. - ISSN 0266-0032 - p. 85 - 100.
farmers’ behaviour - farmers’ intentions - organic materials - soil conservation - soil management - soil organic matter - theory of planned behaviour

Soil organic matter (SOM) is an important resource base for arable farming. For policies on SOM to be effective, insight is needed on why and under which conditions farmers are willing to increase SOM content. This study used the theory of planned behaviour to analyse what prevents or encourages Dutch farmers to increase the SOM content of their fields. In an online survey, 435 arable farmers were asked questions to understand their attitude (perceived benefits), subjective norm (social pressure) and perceived behavioural control (anticipated impediments and obstacles) related to management of SOM. Farmers’ answers were related to their intention to increase SOM content, use of organic materials and perceived increase in SOM content. Our results showed that Dutch farmers are well aware of the possible benefits of SOM content for crop productivity. Farmers’ attitude, subjective norm and perceived decrease in SOM content were significantly related to their intention to increase SOM content. In our farm survey, this intention was very strong: 90% of the farmers stated a high or very high intention to increase the SOM content of their fields. A higher intention to increase SOM content was correlated with the use of organic materials as expressed as total and effective C (P = 0.003 and P = 0.002, respectively), but this did not lead to a perceived increase in SOM content. From a farmer's point of view, this indicates that increasing SOM content is to a large degree beyond their direct influence. The Dutch Manure and Fertiliser Act, costs of organic inputs and the need to cultivate profitable crops (such as potatoes or sugar beet) were indicated as important impeding factors for increasing SOM content.

Open teelten en klimaatadaptatie in relatie tot de financiële weerbaarheid
Verhagen, Jan ; Asseldonk, Marcel van; Pronk, Annette - \ 2018
Wageningen : Wageningen University & Research, Wageningen Plant Research, Busineess unit Agrosystem (Wageningen Plant Research rapport WPR-755) - 33
Guidelines for Biomarker of Food Intake Reviews (BFIRev) : How to conduct an extensive literature search for biomarker of food intake discovery
Praticò, Giulia ; Gao, Qian ; Scalbert, Augustin ; Vergères, Guy ; Kolehmainen, Marjukka ; Manach, Claudine ; Brennan, Lorraine ; Pedapati, Sri Harsha ; Afman, Lydia A. ; Wishart, David S. ; Vázquez-Fresno, Rosa ; Lacueva, Cristina Andres ; Garcia-Aloy, Mar ; Verhagen, Hans ; Feskens, Edith J.M. ; Dragsted, Lars O. - \ 2018
Genes & Nutrition 13 (2018)1. - ISSN 1555-8932
Biomarkers - Food exposure markers - Literature search methodology - Metabolomics - Systematic review
Identification of new biomarkers of food and nutrient intake has developed fast over the past two decades and could potentially provide important new tools for compliance monitoring and dietary intake assessment in nutrition and health science. In recent years, metabolomics has played an important role in identifying a large number of putative biomarkers of food intake (BFIs). However, the large body of scientific literature on potential BFIs outside the metabolomics area should also be taken into account. In particular, we believe that extensive literature reviews should be conducted and that the quality of all suggested biomarkers should be systematically evaluated. In order to cover the literature on BFIs in the most appropriate and consistent manner, there is a need for appropriate guidelines on this topic. These guidelines should build upon guidelines in related areas of science while targeting the special needs of biomarker methodology. This document provides a guideline for conducting an extensive literature search on BFIs, which will provide the basis to systematically validate BFIs. This procedure will help to prioritize future work on the identification of new potential biomarkers and on validating these as well as other biomarker candidates, thereby providing better tools for future studies in nutrition and health.
Serologic evidence of West Nile virus and Usutu virus infections in Eurasian coots in the Netherlands
Lim, S.M. ; Geervliet, M. ; Verhagen, J.H. ; Müskens, G.J.D.M. ; Majoor, F.A. ; Osterhaus, Albert D.M.E. ; Martina, Byron E. - \ 2018
Zoonoses and Public Health 65 (2018)1. - ISSN 1863-1959 - p. 96 - 102.
Reservoir hosts - Surveillance - Usutu virus - Vector-borne diseases - West Nile virus - Wild birds

West Nile virus (WNV) and Usutu virus (USUV) are arboviruses that are maintained in enzootic transmission cycles between mosquitoes and birds and are occasionally transmitted to mammals. As arboviruses are currently expanding their geographic range and emerging in often unpredictable locations, surveillance is considered an important element of preparedness. To determine whether sera collected from resident and migratory birds in the Netherlands as part of avian influenza surveillance would also represent an effective source for proactive arbovirus surveillance, a random selection of such sera was screened for WNV antibodies using a commercial ELISA. In addition, sera of jackdaws and carrion crows captured for previous experimental infection studies were added to the selection. Of the 265 screened serum samples, 27 were found to be WNV-antibody-positive, and subsequent cross-neutralization experiments using WNV and USUV confirmed that five serum samples were positive for only WNV-neutralizing antibodies and seven for only USUV. The positive birds consisted of four Eurasian coots (Fulica atra) and one carrion crow (Corvus corone) for WNV, of which the latter may suggest local presence of the virus, and only Eurasian coots for USUV. As a result, the screening of a small selection of serum samples originally collected for avian influenza surveillance demonstrated a seroprevalence of 1.6% for WNV and 2.8% for USUV, suggesting that this sustained infrastructure could serve as a useful source for future surveillance of arboviruses such as WNV and USUV in the Netherlands.

Mapping the potential (quickscan) for salt and drought tolerant crops and cropping systems in the Mekong Delta
Verhagen, Jan ; Blom-Zandstra, Greet ; Nguyen, Duc Xuan Chuong ; Nguyen, Thi Viet Ha ; Prins, Peter ; Linden, Gerard van der - \ 2017
Wageningen : Wageningen Plant Research, Business Unit Agrosystems Research (Wageningen Plant Research rapport WPR-716) - 15
Data from: Recent natural selection causes adaptive evolution of an avian polygenic trait
Bosse, M. ; Spurgin, Lewis G. ; Laine, Veronika N. ; Cole, Ella F. ; Firth, Josh A. ; Gienapp, Phillip ; Gosler, Andrew G. ; McMahon, Keith ; Poissant, Jocelyn ; Verhagen, I.C. ; Groenen, M. ; Oers, C.H.J. ; Sheldon, Ben C. ; Visser, M.E. ; Slate, Jon - \ 2017
adaptation - evolution - genomics - natural selection - bill length - birds - Parus major
We used extensive data from a long-term study of great tits (Parus major) in the United Kingdom and Netherlands to better understand how genetic signatures of selection translate into variation in fitness and phenotypes. We found that genomic regions under differential selection contained candidate genes for bill morphology and used genetic architecture analyses to confirm that these genes, especially the collagen gene COL4A5, explained variation in bill length. COL4A5 variation was associated with reproductive success, which, combined with spatiotemporal patterns of bill length, suggested ongoing selection for longer bills in the United Kingdom. Last, bill length and COL4A5 variation were associated with usage of feeders, suggesting that longer bills may have evolved in the United Kingdom as a response to supplementary feeding.
A flora of agricultural and horticultural crops : a quick scan of selected crops in the Mekong Delta
Blom-Zandstra, Greet ; Nardelli, Martina ; Duc Xuan Chuong, Nguyen ; Thi Thu Hien, Vu ; Bao Quoc, Nguyen ; Thi Viet Ha, Nguyen ; Linden, Gerard van der; Verhagen, Jan - \ 2017
Lelystad : Wageningen Research Foundation (WR) business units Agrosystems Research and Greenhouse Horticulture (Report / WPR 688) - 53
A scheme for a flexible classification of dietary and health biomarkers
Gao, Qian ; Praticò, G. ; Scalbert, A. ; Vergères, Guy ; Kolehmainen, M. ; Manach, Claudine ; Brennan, L. ; Afman, L.A. ; Wishart, D.S. ; Andres-Lacueva, Cristina ; Garcia-Aloy, M. ; Verhagen, H. ; Feskens, E.J.M. ; Dragsted, L.O. - \ 2017
Genes & Nutrition 12 (2017). - ISSN 1555-8932
Biomarkers are an efficient means to examine intakes or exposures and their biological effects and to assess system susceptibility. Aided by novel profiling technologies, the biomarker research field is undergoing rapid development and new putative biomarkers are continuously emerging in the scientific literature. However, the existing concepts for classification of biomarkers in the dietary and health area may be ambiguous, leading to uncertainty about their application. In order to better understand the potential of biomarkers and to communicate their use and application, it is imperative to have a solid scheme for biomarker classification that will provide a well-defined ontology for the field. In this manuscript, we provide an improved scheme for biomarker classification based on their intended use rather than the technology or outcomes (six subclasses are suggested: food compound intake biomarkers (FCIBs), food or food component intake biomarkers (FIBs), dietary pattern biomarkers (DPBs), food compound status biomarkers (FCSBs), effect biomarkers, physiological or health state biomarkers). The application of this scheme is described in detail for the dietary and health area and is compared with previous biomarker classification for this field of research.
Recent natural selection causes adaptive evolution of an avian polygenic trait
Bosse, Mirte ; Spurgin, Lewis G. ; Laine, Veronika N. ; Cole, Ella F. ; Firth, Josh A. ; Gienapp, Phillip ; Gosler, Andrew G. ; McMahon, Keith ; Poissant, Jocelyn ; Verhagen, Irene ; Groenen, Martien A.M. ; Oers, Kees van; Sheldon, Ben C. ; Visser, Marcel E. ; Slate, Jon - \ 2017
Science 358 (2017)6361. - ISSN 0036-8075 - p. 365 - 368.

We used extensive data froma long-term study of great tits (Parusmajor) in theUnitedKingdom and Netherlands to better understand how genetic signatures of selection translate into variation in fitness and phenotypes.We found that genomic regions under differential selection contained candidate genes for bill morphology and used genetic architecture analyses to confirmthat these genes, especially the collagen gene COL4A5, explained variation in bill length. COL4A5 variation was associated with reproductive success, which, combined with spatiotemporal patterns of bill length, suggested ongoing selection for longer bills in the United Kingdom. Last, bill length and COL4A5 variation were associated with usage of feeders, suggesting that longer bills may have evolved in the United Kingdom as a response to supplementary feeding.

Water-food-energy nexus : A quick scan
Reinhard, Stijn ; Verhagen, Jan ; Wolters, Wouter ; Ruben, Ruerd - \ 2017
Wageningen : Wageningen Economic Research (Wageningen Economic Research menorandum 2017-096) - 23
Approaches aiming at sustainable production
Verhagen, Jan ; Blom, Greet ; Beek, Christy van; Verzandvoort, Simone - \ 2017
Wageningen : Wageningen Plant Research (Report / WPR 677) - 27
sustainable agriculture - organic farming - agroecology - sustainability - duurzame landbouw - biologische landbouw - agro-ecologie - duurzaamheid (sustainability)
This study has two objectives: 1) Look at several approaches that aim to increase or promote sustainable agriculture, and to identify similarities and differences, while deriving implications for policy makers. 2) Provide insight in how the approaches to sustainable agriculture contribute to these priorities.
No evidence that migratory geese disperse avian influenza viruses from breeding to wintering ground
Yin, Shenglai ; Kleijn, David ; Müskens, Gerard J.D.M. ; Fouchier, Ron A.M. ; Verhagen, Josanne H. ; Glazov, Petr M. ; Si, Yali ; Prins, Herbert H.T. ; Boer, Fred de - \ 2017
PLoS One 12 (2017)5. - ISSN 1932-6203 - 11 p.

Low pathogenic avian influenza virus can mutate to a highly pathogenic strain that causes severe clinical signs in birds and humans. Migratory waterfowl, especially ducks, are considered the main hosts of low pathogenic avian influenza virus, but the role of geese in dispersing the virus over long-distances is still unclear. We collected throat and cloaca samples from three goose species, Bean goose (Anser fabalis), Barnacle goose (Branta leucopsis) and Greater white-fronted goose (Anser albifrons), from their breeding grounds, spring stopover sites, and wintering grounds. We tested if the geese were infected with low pathogenic avian influenza virus outside of their wintering grounds, and analysed the spatial and temporal patterns of infection prevalence on their wintering grounds. Our results show that geese were not infected before their arrival on wintering grounds. Barnacle geese and Greater white-fronted geese had low prevalence of infection just after their arrival on wintering grounds in the Netherlands, but the prevalence increased in successive months, and peaked after December. This suggests that migratory geese are exposed to the virus after their arrival on wintering grounds, indicating that migratory geese might not disperse low pathogenic avian influenza virus during autumn migration.

Discordant detection of avian influenza virus subtypes in time and space between poultry and wild birds; Towards improvement of surveillance programs
Verhagen, Josanne H. ; Lexmond, Pascal ; Vuong, Oanh ; Schutten, Martin ; Guldemeester, Judith ; Osterhaus, Albert D.M.E. ; Elbers, Armin R.W. ; Slaterus, Roy ; Hornman, Menno ; Koch, Guus ; Fouchier, Ron A.M. ; Lierz, Michael - \ 2017
PLoS One 12 (2017)3. - ISSN 1932-6203
Avian influenza viruses from wild birds can cause outbreaks in poultry, and occasionally infect humans upon exposure to infected poultry. Identification and characterization of viral reservoirs and transmission routes is important to develop strategies that prevent infection of poultry, and subsequently virus transmission between poultry holdings and to humans. Based on spatial, temporal and phylogenetic analyses of data generated as part of intense and large-scale influenza surveillance programs in wild birds and poultry in the Netherlands from 2006 to 2011, we demonstrate that LPAIV subtype distribution differed between wild birds and poultry, suggestive of host-range restrictions. LPAIV isolated from Dutch poultry were genetically most closely related to LPAIV isolated from wild birds in the Netherlands or occasionally elsewhere in Western Europe. However, a relatively long time interval was observed between the isolations of related viruses from wild birds and poultry. Spatial analyses provided evidence for mallards (Anas platyrhynchos) being more abundant near primary infected poultry farms. Detailed year-round investigation of virus prevalence and wild bird species distribution and behavior near poultry farms should be used to improve risk assessment in relation to avian influenza virus introduction and retarget avian influenza surveillance programs
Trade-offs in soil fertility management on arable farms
Bos, Jules F.F.P. ; Berge, Hein F.M. ten; Verhagen, Jan ; Ittersum, Martin K. van - \ 2017
Agricultural Systems 157 (2017). - ISSN 0308-521X - p. 292 - 302.
Carbon sequestration - Climate change mitigation - Greenhouse gases - Linear programming - Nitrogen losses

Crop production and soil fertility management implies a multitude of decisions and activities on crop choice, rotation design and nutrient management. In practice, the choices to be made and the resulting outcomes are subject to a wide range of objectives and constraints. Objectives are economic as well as environmental, for instance sequestering carbon in agricultural soils or reducing nitrogen losses. Constraints originate from biophysical and institutional conditions that may restrict the possibilities for choosing crops or using specific cultivation and fertilization practices. To explore the consequences of management interventions to increase the supply of organic C to the soil on income and N losses, we developed the linear programming model NutMatch. The novelty of the model is the coherent description of mutual interdependencies amongst a broad range of sustainability indicators related to soil fertility management in arable cropping, enabling the quantification of synergies and trade-offs between objectives. NutMatch was applied to four different crop rotations subjected to four fertiliser strategies differing in the use of the organic fertilisers cattle slurry, pig slurry or compost, next to mineral fertiliser. Each combination of rotation and fertiliser strategy contributed differently to financial return, N emissions and organic matter inputs into the soil.Our model calculations show that, at the rotational level, crop residues, cattle slurry and compost each substantially contributed to SOC accumulation (range 200-450 kg C ha-1 yr-1), while contributions of pig slurry and cover crops were small (20-50 kg C ha-1 yr-1). The use of compost and pig slurry resulted in increases of 0.61-0.73 and 3.15-3.38 kg N2O-N per 100 kg extra SOC accumulated, respectively, with the other fertilizers taking an intermediate position. From a GHG emission perspective, the maximum acceptable increase is 0.75 kg N2O-N per 100 kg extra SOC accumulated, which was only met by compost. Doubling the winter wheat area combined with the cultivation of cover crops to increase SOC accumulation resulted in a net GHG emission benefit, but was associated with a financial trade-off of 2.30-3.30 euro per kg SOC gained.Our model calculations suggest that trade-offs between C inputs and emissions of greenhouse gases (notably N2O) or other pollutants (NO3, NH3) can be substantial. Due to the many data from a large variety of sources incorporated in the model, the trade-offs are uncertain. Our model-based explorations provide insight in soil carbon sequestration options and their limitations vis-a-vis other objectives.

Economic feasibility and climate benefits of using struvite from the Netherlands as a phosphate (P) fertilizer in West Africa
Vries, Sander de; Postma, Romke ; Scholl, Laura van; Blom-Zandstra, Greet ; Verhagen, Jan ; Harms, Imke - \ 2016
Wageningen : Wageningen Plant Research (Wageningen Plant Research report 673) - 47
Not everyone dealing with agricultural issues in Malawi appreciates the fact that smallholders in this country face challenges which are unique in Africa. No other country in Sub-Saharan Africa has population densities of up to 250 to the square km combined with a single rainy season of five months. The five areas with comparable population densities to those of Malawi have rain all the year or have two rainy seasons. Their smallholders can grow perennial food crops and, on a small piece of land, establish high value perennial cash crops such as tea, coffee and vanilla. On the other hand farmers living in areas with comparable rainfall to Malawi occupy comparatively thinly populated countries and so are able to grow larger areas of food crops to meet their needs and larger areas of low value annual cash crops such as cotton and legumes and so raise some cash for their family requirements. Many Malawian smallholders can adopt neither of these options. The overwhelming majority cannot grow coffee or tea because of inadequate rainfall and the limitations of small farm size means that they can only produce small amounts of any crop other than their basic staples. About 15% grow burley tobacco but that number cannot be increased as the crop is already over-produced. A further 10% raise cash from rice, groundnuts, maize and horticultural crops but the great majority have to allocate all of their land to producing food for the family and rely on low paid casual occupations to raise the cash that they need. It is for this reason that the majority of rural Malawians are classified as being below the poverty line. An appreciation of this situation can help in an understanding of the current state of smallholder farming in Malawi and it is hoped that these notes may cast a little more light on the plight of this country’s millions of small scale farmers and provide some indicators as to how best they can be helped.
Rassenmengsel als risicomanagement
Groten, Jos - \ 2016
Designing eco-effective reverse logistics networks
Souza, Vitor De; Borsato, Milton ; Bloemhof-Ruwaard, Jacqueline - \ 2016
In: Transdisciplinary Engineering: Crossing Boundaries. - IOS Press (Advances in Transdisciplinary Engineering ) - ISBN 9781614997023 - p. 851 - 860.
Industrial Ecology - Material Flow - Reverse Logistics Network Design - TRIZ - Upcycle

Reverse Logistics Networks (RLNs) have grown in importance after return policies became compulsory. Lately, questions have been raised whether they are as helpful to the environment as possible. Efforts have been conducted to optimize RLNs in terms of their eco-efficiency, minimizing costs and emissions; still, results are not advancing with the necessary speed. Alternatively, the ecoeffectiveness ("doing the right thing" for the environment) approach emerges, promoting a supportive relationship, balancing environment and economy. This research aims to model the design (or redesign) process of eco-effective RLNs. There are numerous ecodesign tools focusing on product or service design, but an eco-effective design process conceived specifically for logistics network design purposes has yet to be delivered. Research was carried out using the Design Science Research Methodology and an exemplification was outlined to demonstrate how the process unrolls. The model was conceived using a combination of the TRIZ method, Upcycling and Industrial Symbiosis. Ecoefficiency of these networks was not evaluated. The proposed design process model will help the conception of more innovative, eco-effective logistics networks.

Lack of virological and serological evidence for continued circulation of highly pathogenic avian influenza H5N8 virus in wild birds in the Netherlands, 14 November 2014 to 31 January 2016
Poen, M.J. ; Verhagen, J.H. ; Manvell, R.J. ; Brown, I. ; Bestebroer, T.M. ; Vliet, S. van der; Vuong, O. ; Scheuer, R.D. ; Jeugd, H.P. van der; Nolet, B.A. ; Kleyheeg, E. ; Müskens, G.J.D.M. ; Majoor, F.A. ; Grund, C. ; Fouchier, Ron A.M. - \ 2016
EuroSurveillance 21 (2016)38. - ISSN 1025-496X - 11 p.

In 2014, H5N8 clade highly pathogenic avian influenza (HPAI) viruses of the A/Goose/ Guangdong/1/1996 lineage emerged in poultry and wild birds in Asia, Europe and North America. Here, wild birds were extensively investigated in the Netherlands for HPAI H5N8 virus (real-time polymerase chain reaction targeting the matrix and H5 gene) and antibody detection (haemagglutination inhibition and virus neutralisation assays) before, during and after the first virus detection in Europe in late 2014. Between 21 February 2015 and 31 January 2016, 7,337 bird samples were tested for the virus. One HPAI H5N8 virus-infected Eurasian wigeon (Anas penelope) sampled on 25 February 2015 was detected. Serological assays were performed on 1,443 samples, including 149 collected between 2007 and 2013, 945 between 14 November 2014 and 13 May 2015, and 349 between 1 September and 31 December 2015. Antibodies specific for HPAI H5 clade were absent in wild bird sera obtained before 2014 and present in sera collected during and after the HPAI H5N8 emergence in Europe, with antibody incidence declining after the 2014/15 winter. Our results indicate that the HPAI H5N8 virus has not continued to circulate extensively in wild bird populations since the 2014/15 winter and that independent maintenance of the virus in these populations appears unlikely.

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