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In vitro fermentation techniques (IVFT) have been widely used to evaluate the nutritive value of feeds for ruminants and in the last decade to assess the effect of different nutritional strategies on methane (CH4) production. However, many technical factors may influence the results obtained. The present review has been prepared by the 'Global Network' FACCE-JPI international research consortium to provide a critical evaluation of the main factors that need to be considered when designing, conducting and interpreting IVFT experiments that investigate nutritional strategies to mitigate CH emission from ruminants. Given the increasing and wide-scale use of IVFT, there is a need to critically review reports in the literature and establish what criteria are essential to the establishment and implementation of in vitro techniques. Key aspects considered include: i) donor animal species and number of animal used, ii) diet fed to donor animals, iii) collection and processing of rumen fluid as inoculum, iv) choice of substrate and incubation buffer, v) incubation procedures and CH measurements, vi) headspace gas composition and vii) comparability of in vitro and in vivo measurements. Based on an evaluation of experimental evidence, a set of technical recommendations are presented to harmonize IVFT for feed evaluation, assessment of rumen function and CH production., This review is part of the FACCE-JPI ‘Global Network’ project and the ‘Feeding and Nutrition Network’ of Livestock Research Group, in the Global Research Alliance for Agricultural Greenhouse Gases (www.globalresearchalliance.org). Authors acknowledge national funding from INIA (Spain, project MIT01-GLOBALNET-EEZ), the Ministry of Economic Affairs (The Netherlands; project Global Research Alliance on Agricultural Greenhouses Gases, BO-20-007-006), USDA-NIFA (USA), French National Research Agency through the program FACCE-JPI program, the Irish contribution to this JPI-FACCE project was funded by the Agricultural GHG Research Initiative for Ireland (AGRI-I), Department for Environment Food & Rural Affairs (UK), BLW (Switzerland) and Academy of Finland, Helsinki, Finland (Project 281337).

Ruminant husbandry is a major source of anthropogenic greenhouse gases (GHG). Filling knowledge gaps and providing expert recommendation are important for defining future research priorities, improving methodologies and establishing science-based GHG mitigation solutions to government and non-governmental organisations, advisory/extension networks, and the ruminant livestock sector. The objectives of this review is to summarize published literature to provide a detailed assessment of the methodologies currently in use for measuring enteric methane (CH) emission from individual animals under specific conditions, and give recommendations regarding their application. The methods described include respiration chambers and enclosures, sulphur hexafluoride tracer (SF) technique, and techniques based on short-term measurements of gas concentrations in samples of exhaled air. This includes automated head chambers (e.g. the GreenFeed system), the use of carbon dioxide (CO) as a marker, and (handheld) laser CH detection. Each of the techniques are compared and assessed on their capability and limitations, followed by methodology recommendations. It is concluded that there is no ‘one size fits all’ method for measuring CH emission by individual animals. Ultimately, the decision as to which method to use should be based on the experimental objectives and resources available. However, the need for high throughput methodology e.g. for screening large numbers of animals for genomic studies, does not justify the use of methods that are inaccurate. All CH measurement techniques are subject to experimental variation and random errors. Many sources of variation must be considered when measuring CH concentration in exhaled air samples without a quantitative or at least regular collection rate, or use of a marker to indicate (or adjust) for the proportion of exhaled CH sampled. Consideration of the number and timing of measurements relative to diurnal patterns of CH emission and respiratory exchange are important, as well as consideration of feeding patterns and associated patterns of rumen fermentation rate and other aspects of animal behaviour. Regardless of the method chosen, appropriate calibrations and recovery tests are required for both method establishment and routine operation. Successful and correct use of methods requires careful attention to detail, rigour, and routine self-assessment of the quality of the data they provide., This review is part of the FACCE-JPI ‘Global Network’ project. Authors acknowledge national funding from the Departmentfor Environment, Food and Rural Affairs (UK), INIA (Spain, project MIT01-GLOBALNET-EEZ), the Ministry of Economic Affairs(The Netherlands; project Global Research Alliance on Agricultural Greenhouses Gases, BO-20-007-006), USDA-NIFA (USA),French National Research Agency through the FACCE-JPI program, Agricultural GHG Research Initiative for Ireland (AGRI-I),Academy of Finland, Helsinki, Finland (Project 281337), and BLW (Switzerland).

The objective of this study was to evaluate the performance of existing models predicting enteric methane (CH) emissions, using a large database (3183 individual data from 103 in vivo studies on dairy and beef cattle, sheep and goats fed diets from different countries). The impacts of dietary strategies to reduce CH emissions, and of diet quality (described by organic matter digestibility (dOM) and neutral-detergent fiber digestibility (dNDF)) on model performance were assessed by animal category. The models were first assessed based on the root mean square prediction error (RMSPE) to standard deviation of observed values ratio (RSR) to account for differences in data between models and then on the RMSPE. For dairy cattle, the CH (g/d) predicting model based on feeding level (dry matter intake (DMI)/body weight (BW)), energy digestibility (dGE) and ether extract (EE) had the smallest RSR (0.66) for all diets, as well as for the high-EE diets (RSR = 0.73). For mitigation strategies based on lowering NDF or improving dOM, the same model (RSR = 0.48 to 0.60) and the model using DMI and neutral- and acid-detergent fiber intakes (RSR = 0.53) had the smallest RSR, respectively. For diets with high starch (STA), the model based on nitrogen, ADF and STA intake presented the smallest RSR (0.84). For beef cattle, all evaluated models performed moderately compared with the models of dairy cattle. The smallest RSR (0.83) was obtained using variables of energy intake, BW, forage content and dietary fat, and also for the high-EE and the low-NDF diets (RSR = 0.84 to 0.86). The IPCC Tier 2 models performed better when dietary STA, dOM or dNDF were high. For sheep and goats, the smallest RSR was observed from a model for sheep based on dGE intake (RSR = 0.61). Both IPCC models had low predictive ability when dietary EE, NDF, dOM and dNDF varied (RSR = 0.57 to 1.31 in dairy, and 0.65 to 1.24 in beef cattle). The performance of models depends mostly on explanatory variables and not on the type of data (individual vs. treatment means) used in their development or evaluation. Some empirical models give satisfactory prediction error compared with the error associated with measurement methods. For better prediction, models should include feed intake, digestibility and additional information on dietary concentrations of EE and structural and nonstructural carbohydrates to account for different dietary mitigating strategies., This study is part of the Joint Programming Initiative on Agriculture, Food Security and Climate Change (FACCE-JPI)’s “GLOBAL NETWORK” project and the “Feeding and Nutrition Network” (http://animalscience.psu.edu/fnn) of the Livestock Research Group within the Global Research Alliance for Agricultural Greenhouse Gases (www.globalresearchalliance.org). Authors gratefully acknowledge funding for this project from: USDA National Institute of Food and Agriculture (Grant no. 2014-67003-21979) University of California, Davis Sesnon Endowed Chair Program, USDA, and Austin Eugene Lyons Fellowship (University of California, Davis); Funding from USDA National Institute of Food and Agriculture Federal Appropriations under Project PEN 04539 and Accession number 1000803, DSM Nutritional Products (Basel, Switzerland), Pennsylvania Soybean Board (Harrisburg, PA, USA), Northeast Sustainable Agriculture Research and Education (Burlington, VT, USA), and PMI Nutritional Additives (Shoreview, MN, USA); the Ministry of Economic Affairs (the Netherlands; project BO-20-007-006; Global Research Alliance on Agricultural Greenhouse Gases), the Product Board Animal Feed (Zoetermeer, the Netherlands) and the Dutch Dairy Board (Zoetermeer, the Netherlands); The Cofund for Monitoring & Mitigation of Greenhouse Gases from Agri- and Silvi-culture (FACCE ERA-GAS)’s project Capturing Effects of Diet on Emissions from Ruminant Systems and the Dutch Ministry of Agriculture, Nature and Food Quality (AF-EU-18010 & BO-4400159-01); USDA National Institute of Food and Agriculture (Hatch Multistate NC-1042 Project Number NH00616-R; Project Accession Number 1001855) and the New Hampshire Agricultural Experiment Station (Durham, NH); French National Research Agency through the FACCE-JPI program (ANR-13-JFAC-0003-01); the Department of Agriculture, Food and the Marine, Ireland Agricultural GHG Research Initiative for Ireland (AGRI-I) project; Academy of Finland (No. 281337), Helsinki, Finland; Swiss Federal Office of Agriculture, Berne, Switzerland; the Department for Environment, Food and Rural Affairs (Defra; UK); Defra, the Scottish Government, DARD, and the Welsh Government as part of the UK’s Agricultural GHG Research Platform projects (www.ghgplatform.org.uk); INIA (Spain, project MIT01-GLOBALNET-EEZ); German Federal Ministry of Food and Agriculture (BMBL) through the Federal Office for Agriculture and Food (BLE); Swedish Infrastructure for Ecosystem Science (SITES) at Röbäcksdalen Research Station; Comisión Nacional de Investigación Científica y Tecnológica, Fondo Nacional de Desarrollo Científico y Tecnológico (Grant Nos. 11110410 and 1151355) and Fondo Regional de Tecnología Agropecuaria (FTG/RF-1028-RG); European Commission through SMEthane (FP7-SME-262270).

State-of-the-art water-oxidation catalysts (WOCs) in acidic electrolytes usually contain expensive noble metals such as ruthenium and iridium. However, they too expensive to be implemented broadly in semiconductor photoanodes for photoelectrochemical (PEC) water splitting devices. Here, an Earth-abundant CoFe Prussian blue analogue (CoFe-PBA) is incorporated with core–shell Fe2O3/Fe2TiO5 type II heterojunction nanowires as composite photoanodes for PEC water splitting. Those deliver a high photocurrent of 1.25 mA cm−2 at 1.23 V versus reversible reference electrode in acidic electrolytes (pH = 1). The enhancement arises from the synergic behavior between the successive decoration of the hematite surface with nanolayers of Fe2TiO5 and then, CoFe-PBA. The underlying physical mechanism of performance enhancement through formation of the Fe2O3/Fe2TiO5/ CoFe-PBA heterostructure reveals that the surface states’ electronic levels of hematite are modified such that an interfacial charge transfer becomes kinetically favorable. These findings open new pathways for the future design of cheap and efficient hematite-based photoanodes in acidic electrolytes., This work was supported by the Spanish Ministerio de Economia y Competitividad (MINECO, Grants CTQ2015-71287-R, CTQ2015-71287-R, and CTQ2015-68770-R) and the coordinated Project ValPEC (ENE2017-85087-C3), the BIST Ignite Project inWOC2 and the Generalitat de Catalunya (2017 SGR 90, 2017 SGR 327, 2017 SGR 329, 2017 SGR 1246, and 2017 SGR 1406). ICN2 acknowledges the support from the Severo Ochoa Program (MINECO, Grant SEV-2017-0706). ICN2, ICIQ, and IREC are funded by the CERCA Programme/Generalitat de Catalunya. P.Y.T. acknowledges the scholarship support of DAAD short term grant. H.C.D. acknowledges support from the Deutsche Forschungsgemeinschaft (SFB 917). F.S.H. thanks the “LaCaixa”-Severo Ochoa International Programme (Programa internacional de Becas “LaCaixa”- Severo Ochoa) for a Ph.D. fellowship. P.P. and M.H. thank for the support by the Federal Ministry for Economic Affairs and Energy (BMWi) (Fundingregistration number: 03ET6080E). M.L. thanks the COST Action StableNextSol project MP1307, supported by COST (European Cooperation in Science and Technology). H.X. acknowledges the Spanish MINECO through the Severo Ochoa Centers of Excellence Program under Grant SEV-2013-0295 for the postdoctoral contract.

Over the past 50 years, the population of Continental Black-tailed Godwits Limosa limosa limosa breeding of the East Atlantic Flyway has been in steep decline. This decline has previously been documented in trend analyses and six Netherlands-wide count-based population estimates, the last of which was completed in 1999. We provide an updated population size estimate and describe inter-annual fluctuations in the population between 2007 and 2015. To generate these estimates, we integrated a mark-recapture survival analysis with estimates of the densities of colour-marked individuals at migratory staging sites with known proportions of Continental and Icelandic L. l. islandica Black-tailed Godwits within a Bayesian framework. The use of these analytical techniques means that, in contrast with earlier efforts, our estimates are accompanied with confidence intervals, allowing us to estimate the population size with known precision. Using additional information on the breeding destination of 43 godwits equipped with satellite transmitters at Iberian staging areas, we found that 87% (75-95% 95% CI) of the nominate subspecies in the East Atlantic Flyway breed in The Netherlands. We estimated that the number of breeding pairs in The Netherlands has declined from 47,000 (38,000-56,000) pairs in 2007 to 33,000 (26,000-41,000) in 2015. Despite a temporary increase in 2010 and 2011, the population declined by an average of 3.7% per year over the entire period from 2007-2015, and by 6.3% from 2011-2015. We conclude that investing in an intensive demographic programme at a regional scale, when combined with targeted resightings of marked individuals elsewhere, can yield population estimates at the flyway scale., This study was funded mainly by the former Netherlands Ministry of Agriculture, Nature Management and Food Safety, now subsumed in the Ministry of Economic Affairs, by the Province of Fryslân, and by the Spinoza Premium Award 2014 from The Netherlands Organization for Scientific Research (NWO) to TP, with some additional funding by the Prins Bernard Cultuurfonds, the Van der Hucht Beukelaar Stichting, BirdLife Netherlands and WWF-Netherlands through Global Flyway Network and the Chair in Flyway Ecology, FP7-Regpot project ECOGENES (Grant No. 264125), the NWO-TOP grant ‘Shorebirds in space’ (854.11.004) awarded to TP, ExpeER Ecosystem Research, ‘ICTS-RBD’ to the ESFRI LifeWatch, MINECO, and European Union Structural Funds (AIC-A2011-0706).

Mass-flowering crops (MFCs) are increasingly cultivated and might influence pollinator communities in MFC fields and nearby semi-natural habitats (SNHs). Across six European regions and 2 years, we assessed how landscape-scale cover of MFCs affected pollinator densities in 408 MFC fields and adjacent SNHs. In MFC fields, densities of bumblebees, solitary bees, managed honeybees and hoverflies were negatively related to the cover of MFCs in the landscape. In SNHs, densities of bumblebees declined with increasing cover of MFCs but densities of honeybees increased. The densities of all pollinators were generally unrelated to the cover of SNHs in the landscape. Although MFC fields apparently attracted pollinators from SNHs, in landscapes with large areas of MFCs they became diluted. The resulting lower densities might negatively affect yields of pollinator-dependent crops and the reproductive success of wild plants. An expansion of MFCs needs to be accompanied by pollinator-supporting practices in agricultural landscapes., The study was funded by the EU FP7 project “Status and Trends of European Pollinators” (244 090, www.STEP-project.net) and Biodiversa‐FACCE project “Enhancing biodiversity‐based ecosystem services to crops through optimised densities of green infrastructure in agricultural landscapes” (PCIN‐2014‐048, http://www.cec.lu.se/research/ecodeal). JPGV and MV acknowledge support from the Severo Ochoa Program for Centres of Excellence in R+D+I (SEV‐2012‐0262), and FLORMAS (CGL 2012‐33801). JBW, VJW, SGP and SPMR acknowledge the support of Insect Pollinators Initiative research funded jointly by a grant from BBSRC, Defra, NERC, the Scottish Government and the Wellcome Trust, under the Living with Environmental Change Partnership. HGS and RB were supported by the Swedish Research Council FORMAS. JS and DK were supported by the Dutch Ministry of Economic Affairs (BO‐11‐011.01‐011 and KB‐14‐003‐006), AH and ISD were supported by the DFG Collaborative Research Centre 1047 Insect timing, and ISD and MD through the 2013–2014 BiodivERsA/FACCE – JPI project Ecodeal with national funding from BMBF.

In literature, varying and sometimes conflicting effects of physicochemical properties of nanoparticles (NPs) are reported on their uptake and effects in organisms. To address this, small- and medium-sized (20 and 50 nm) silver nanoparticles (AgNPs) with specified different surface coating/charges were synthesized and used to systematically assess effects of NP-properties on their uptake and effects in vitro. Silver nanoparticles were fully characterized for charge and size distribution in both water and test media. Macrophage cells (RAW 264.7) were exposed to these AgNPs at different concentrations (0-200 µg/ml). Uptake dynamics, cell viability, induction of tumor necrosis factor (TNF)-α, ATP production, and reactive oxygen species (ROS) generation were assessed. Microscopic imaging of living exposed cells showed rapid uptake and subcellular cytoplasmic accumulation of AgNPs. Exposure to the tested AgNPs resulted in reduced overall viability. Influence of both size and surface coating (charge) was demonstrated, with the 20-nm-sized AgNPs and bovine serum albumin (BSA)-coated (negatively charged) AgNPs being slightly more toxic. On specific mechanisms of toxicity (TNF-α and ROS production) however, the AgNPs differed to a larger extent. The highest induction of TNF-α was found in cells exposed to the negatively charged AgNP_BSA, both sizes (80× higher than control). Reactive oxygen species induction was only significant with the 20 nm positively charged AgNP_Chit., This work was financially supported by NanoNextNL, a micro- and nanotechnology consortium of the Government of The Netherlands and 130 partners; funding was also received from Managing Risks of Nanoparticles, MARINA (EU-FP7, contract CP-FP 263215); and the Strategic Research Funds titled Novel technologies by the Ministry of Economic Affairs of The Netherlands. Synthesis and characterization of the AgNPs used in this study received support from the QualityNano Project (http://www.qualitynano.eu/) financed by the European Community Research Infrastructures under the FP7 Capacities Program (Grant Number INFRA-2010-262163).

The GMO Risk Assessment and Communication of Evidence (GRACE; www.grace-fp7.eu) project is funded by the European Commission within the 7th Framework Programme. A key objective of GRACE is to conduct 90-day animal feeding trials, animal studies with an extended time frame as well as analytical, in vitro and in silico studies on genetically modified (GM) maize in order to comparatively evaluate their use in GM plant risk assessment. In the present study, the results of two 90-day feeding trials with two different GM maize MON810 varieties, their near-isogenic non-GM varieties and four additional conventional maize varieties are presented. The feeding trials were performed by taking into account the guidance for such studies published by the EFSA Scientific Committee in 2011 and the OECD Test Guideline 408. The results obtained show that the MON810 maize at a level of up to 33 % in the diet did not induce adverse effects in male and female Wistar Han RCC rats after subchronic exposure, independently of the two different genetic backgrounds of the event., This study was carried out as part of the GRACE project (“GMO Risk Assessment and Communication of Evidence”), financially supported by the 7th Framework Programme of the European Community for Research, Technological Development and Demonstration Activities (FP7), Grant Agreement No. 311957, as well as the Dutch Ministry of Economic Affairs and various other co-sponsors.

While self-assembled molecular building blocks could lead to many next-generation functional organic nanomaterials, control over the thin-film morphologies to yield monolithic sub-5 nm patterns with 3D orientational control at macroscopic length scales remains a grand challenge. A series of photoresponsive hybrid oligo(dimethylsiloxane) liquid crystals that form periodic cylindrical nanostructures with periodicities between 3.8 and 5.1 nm is studied. The liquid crystals can be aligned in-plane by exposure to actinic linearly polarized light and out-of-plane by exposure to actinic unpolarized light. The photoalignment is most efficient when performed just under the clearing point of the liquid crystal, at which the cylindrical nanostructures are reoriented within minutes. These results allow the generation of highly ordered sub-5 nm patterns in thin films at macroscopic length scales, with control over the orientation in a noncontact fashion., This research was supported by the Dutch Technology Foundation STW, which is part of the Netherlands Organisation for Scientific Research (NWO), and which is partly funded by the Ministry of Economic Affairs. C.S.-S. thanks the Spanish MINECO projects MAT2011-27978-C02-02, SAF2014-54763-C2-2-R, Gobierno de Aragón, and FEDER (EU). P.L. is a Senior Research Associate from FRS-FNRS (Belgium). The X-ray diffractometer was financed by The Netherlands Organisation for Scientific Research (NWO).

et al., Physicochemical properties of nanoparticles influence their environmental fate and toxicity, and studies investigating this are vital for a holistic approach towards a comprehensive and adequate environmental risk assessment. In this study, we investigated the effects of size, surface coating (charge) of silver nanoparticles (AgNPs) – a most commonly-used nanoparticle-type, on the bioaccumulation in, and toxicity (survival, growth, cocoon production) to the earthworm Lumbricus rubellus. AgNPs were synthesized in three sizes: 20, 35 and 50 nm. Surface-coating with bovine serum albumin (AgNP_BSA), chitosan (AgNP_Chit), or polyvinylpyrrolidone (AgNP_PVP) produced negative, positive and neutral particles respectively. In a 28-day sub-chronic reproduction toxicity test, earthworms were exposed to these AgNPs in soil (0–250 mg Ag/kg soil DW). Earthworms were also exposed to AgNO3 at concentrations below known EC50. Total Ag tissue concentration indicated uptake by earthworms was generally highest for the AgNP_BSA especially at the lower exposure concentration ranges, and seems to reach a plateau level between 50 and 100 mg Ag/kg soil DW. Reproduction was impaired at high concentrations of all AgNPs tested, with AgNP_BSA particles being the most toxic. The EC50 for the 20 nm AgNP_BSA was 66.8 mg Ag/kg soil, with exposure to AgNP_PVP (neutral) > Chitosan (positive). Size had an influence on uptake and toxicity of the AgNP_PVP, but not for AgNP_BSA nor AgNP_Chit. This study provides essential information on the role of physicochemical properties of AgNPs in influencing uptake by a terrestrial organism L. rubellus under environmentally relevant conditions. It also provides evidence of the influence of surface coating (charge) and the limited effect of size in the range of 20–50 nm, in driving uptake and toxicity of the AgNPs tested., This work was financially supported by NanoNextNL, a micro- and nano-technology consortium of the Government of The Netherlands and 130 partners; funding was also received from Managing Risks of Nanoparticles, MARINA (EU-FP7, contract CP-FP 263215), and; the Strategic Research Funds titled Novel technologies by the Ministry of Economic Affairs of The Netherlands. Synthesis and characterization of the AgNPs used in this study received support from the QualityNano Project http://www.qualitynano.eu/which is financed by the European Community Research Infrastructures under the FP7 Capacities Programme (Grant No. INFRA-2010-262163).

4 pags., 5 figs., The emission from nanowires can couple to waveguide modes supported by the nanowire geometry, thus governing the far-field angular pattern. To investigate the geometry-induced coupling of the emission to waveguide modes, we acquire Fourier microscopy images of the photoluminescence of nanowires with diameters ranging from 143 to 208 nm. From the investigated diameter range, we conclude that a few nanometers difference in diameter can abruptly change the coupling of the emission to a specific mode. Moreover, we observe a diameter-dependent width of the Gaussian-shaped angular pattern in the far-field emission. This dependence is understood in terms of interference of the guided modes, which emit at the end facets of the nanowire. Our results are important for the design of quantum emitters, solid state lighting, and photovoltaic devices based on nanowires. VC 2016 AIP Publishing LLC, This research was supported by the Dutch technology foundation STW, which is part of the “Netherlands Organisation for Scientific Research (NWO),” and partially funded by the Dutch Ministry of Economic Affairs. This work was also part of the research program of the “Foundation for Fundamental Research on Matter (FOM),” which is financially supported by NWO. J.A.S.G. acknowledges the Spanish Ministerio de Economía y Competitividad for financial support through the Grant Nos. NANOPLASþ (FIS2012-31070) and LENSBEAM (FIS2015-69295-C3-2-P).

Following protection measures implemented since the 1970s, large carnivores are currently increasing in number and returning to areas from which they were absent for decades or even centuries. Monitoring programmes for these species rely extensively on non-invasive sampling and genotyping. However, attempts to connect results of such studies at larger spatial or temporal scales often suffer from the incompatibility of genetic markers implemented by researchers in different laboratories. This is particularly critical for long-distance dispersers, revealing the need for harmonized monitoring schemes that would enable the understanding of gene flow and dispersal dynamics. Based on a review of genetic studies on grey wolves Canis lupus from Europe, we provide an overview of the genetic markers currently in use, and identify opportunities and hurdles for studies based on continent-scale datasets. Our results highlight an urgent need for harmonization of methods to enable transnational research based on data that have already been collected, and to allow these data to be linked to material collected in the future. We suggest timely standardization of newly developed genotyping approaches, and propose that action is directed towards the establishment of shared single nucleotide polymorphism panels, next-generation sequencing of microsatellites, a common reference sample collection and an online database for data exchange. Enhanced cooperation among genetic researchers dealing with large carnivores in consortia would facilitate streamlining of methods, their faster and wider adoption, and production of results at the large spatial scales that ultimately matter for the conservation of these charismatic species., The authors received financial support from the Dutch Ministry for Economic Affairs, the Swiss Federal Office for the Environment, the EuroNatur International Fund for Animal Welfare, Wolves and Humans Foundation, an IF Research contract from the Portuguese Science Foundation (FCT, IF/564/2012), grant SAW‐2011‐SGN‐3 of the Leibniz Association (Germany) and grant DEC‐2014/12/S/NZ8/00624 of the National Science Centre (Poland).

Infections of domestic and wild birds with low-pathogenic avian influenza viruses (LPAIVs) have been associated with protective immunity to subsequent infection. However, the degree and duration of immunity in wild birds from previous LPAIV infection, by the same or a different subtype, are poorly understood. Therefore, we inoculated H13N2 (A/black-headed gull/Netherlands/7/2009) and H16N3 (A/black-headed gull/Netherlands/26/2009) LPAIVs into black-headed gulls (Chroicocephalus ridibundus), their natural host species, and measured the long-term immune response and protection against one or two reinfections over a period of >1 year. This is the typical interval between LPAIV epizootics in wild birds. Reinfection with the same virus resulted in progressively less virus excretion, with complete abrogation of virus excretion after two infections for H13 but not H16. However, reinfection with the other virus affected neither the level nor duration of virus excretion. Virus excretion by immunologically naive birds did not differ in total levels of excreted H13 or H16 virus between first- and second-year birds, but the duration of H13 excretion was shorter for second-year birds. Furthermore, serum antibody levels did not correlate with protection against LPAIV infection. LPAIV-infected gulls showed no clinical signs of disease. These results imply that the epidemiological cycles of H13 and H16 in black-headed gulls are relatively independent from each other and depend mainly on infection of firstyear birds., U.H. was partly funded by a Netherlands Genomics Institute/Virgo Consortium visiting scientist stipend. This work was sponsored by grants from the Dutch Ministry of Economic Affairs, European Research Council project FLUPLAN (250136), and NIH NIAID contracts HHSN266200700010C (2007-2014) and HHSN272201400008C (2014- 2021).

5 pags, 4 figs. -- PACS numbers: 78.68.+m, 73.20.Mf, 73.50.Bk, We map the complex electric fields associated with the scattering of surface plasmon polaritons by single subwavelength holes of different sizes in thick gold films. We identify and quantify the different modes associated with this event, including a radial surface wave with an angularly isotropic amplitude. This wave is shown to arise from the out-of-plane electric dipole induced in the hole, and we quantify the corresponding polarizability, which is in excellent agreement with electromagnetic theory. Time-resolved measurements reveal a time delay of 38±18fs between the surface plasmon polariton and the radial wave, which we attribute to the interaction with a broad hole resonance. © 2012 American Physical Society., This work is part of the research program of the Stichting voor Fundamenteel Onderzoek der Materie (FOM), which is financially supported by the Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO). This work is supported by NanoNextNL of the Dutch ministry EL&I and 130 partners. F. J. G. d. A. acknowledges support from the Spanish MEC (MAT2010-14885 and Consolider NanoLight.es).

Black-headed gulls (Chroicocephalus ridibundus) are a suitable host species to study the epidemiology of low-pathogenic avian influenza virus (LPAIV) infection in wild waterbirds because they are a common colony-breeding species in which LPAIV infection is detected frequently, limited mainly to the H13 and H16 subtypes. However, the sites of virus replication and associated lesions are poorly understood. We therefore performed virological and pathological analyses on tissues of black-headed gulls naturally infected with LPAIV. We found that 24 of 111 black-headed gulls collected from breeding colonies were infected with LPAIV (10 birds with H16N3, one bird with H13N8, 13 birds undetermined), based on virus and viral genome detection in pharyngeal and cloacal swabs. Of these 24 gulls, 15 expressed virus antigen in their tissues. Virus antigen expression was limited to epithelial cells of intestine and cloacal bursa. No histological lesions were detected in association with virus antigen expression. Our findings show that LPAIV replication in the intestinal tract of black-headed gulls is mainly a superficial infection in absence of detectable lesions, as determined recently for natural LPAIV infection in free-living mallards (Anas platyrhynchos). These findings imply that LPAIV in black-headed gulls has adapted to minimal pathogenicity to its host and that potentially the primary transmission route is faecal-oral., J.H.V. and R.A.M.F. are supported by contracts with the Dutch Ministry of Agriculture and NIAID/NIH (number HHSN266200700010C). U.H. had a short-term visiting scientist “José Castillejo” fellowship from the Spanish Ministry of Innovation and Research (Ministerio de Innovación e Investigación MICINN, JC2010-0257) at Erasmus Medical Center. T. Kuiken is supported by contracts with the European Commission through the ANTIGONE project (number 278976) and with the Dutch Ministry of Economic Affairs through the Impulse Veterinary Avian Influenza Research.