Role of position 627 of PB2 and the multibasic cleavage site of the hemagglutinin in the virulence of H5N1 avian influenza virus in chickens and ducks

Highly pathogenic H5N1 avian influenza viruses have caused major disease outbreaks in domestic and free-living birds with transmission to humans resulting in 59% mortality amongst 564 cases. The mutation of the amino acid at position 627 of the viral polymerase basic-2 protein (PB2) from glutamic acid (E) in avian isolates to lysine (K) in human isolates is frequently found, but it is not known if this change affects the fitness and pathogenicity of the virus in birds. We show here that horizontal transmission of A/Vietnam/1203/2004 H5N1 (VN/1203) virus in chickens and ducks was not affected by the change of K to E at PB2-627. All chickens died between 21 to 48 hours post infection (pi), while 70% of the ducks survived infection. Virus replication was detected in chickens within 12 hours pi and reached peak titers in spleen, lung and brain between 18 to 24 hours for both viruses. Viral antigen in chickens was predominantly in the endothelium, while in ducks it was present in multiple cell types, including neurons, myocardium, skeletal muscle and connective tissues. Virus replicated to a high titer in chicken thrombocytes and caused upregulation of TLR3 and several cell adhesion molecules, which may explain the rapid virus dissemination and location of viral antigen in endothelium. Virus replication in ducks reached peak values between 2 and 4 days pi in spleen, lung and brain tissues and in contrast to infection in chickens, thrombocytes were not involved. In addition, infection of chickens with low pathogenic VN/1203 caused neuropathology, with E at position PB2-627 causing significantly higher infection rates than K, indicating that it enhances virulence in chickens.

Invertebrate food sources for waterbirds provided by the reconstructed wetland of Nyirkai-Hany, northwestern Hungary

The Nyirkai-Hany wetland reconstruction area in northwestern Hungary is now designated as a Ramsar and a Natura 2000 site. It was created in 2001–2002 by the Fertő-Hanság National Park Directorate to restore a part of the formerly drained large wetland called Hanság and to offer waterbirds a suitable habitat for feeding and breeding. We focused on this aim of the restoration project and studied the temporal and spatial variation in abundance of birds and their invertebrate prey in this newly created wetland. From April 2007 until May 2008, we sampled plankton, nekton and benthos of different habitats monthly and monitored waterbirds weekly on the three different areas of the Nyirkai-Hany. During our investigations, 135 invertebrate and 53 waterbird species were recorded. Benthos and macrophyte decomposition accelerating guilds were the most abundant waterbird guilds—besides the dominant grazing importer material transporter guild, represented primarily by geese—in the Nyirkai-Hany. Zooplankton assemblages primarily consisted of small species not easily used as a food by planktivorous waterbirds. The low density of zoobenthic biomass and the small extent of shallow water mudflats probably accounted for the scarcity of the bioturbing guild group of birds. Nektonic biomass varied greatly among locations having different vegetation types, was greatest in the shallow water areas dominated by Typha, Carex and Phragmites species and lowest at offshore vegetation-free sites. Chironomids, mayflies and odonates were especially abundant and their biomass significantly correlated with several waterbird species, mainly belonging to the macrophyte decomposition accelerating guild (e.g. Anas platyrynchos, Fulica atra). This guild itself, which has increased in abundance in recent years, showed an exceptionally strong correlation with odonate abundance. These results indicate the growing importance of the Nyirkai-Hany wetland area as a foraging site for waterbirds.

Effects of alterations of primer-binding site sequences on human immunodeficiency virus type 1 replication

The human immunodeficiency virus type 1 genomic RNA primer-binding site (PBS) sequence comprises 18 nucleotides which are complementary to those at the 3' end of the replication initiation primer tRNA(3Lys). To investigate the role of the PBS in viral replication, we either deleted the original wild-type PBS (complementary to tRNA(3Lys) or replaced it with DNA sequences complementary to either tRNA(1,2Lys) or tRNA(Phe). Transfection of COS cells with such molecular constructs yielded similar levels of viral progeny that were indistinguishable with regard to viral proteins and tRNA content. Virus particles derived from PBS-deleted molecular clones were noninfectious for MT-4, Jurkat, and CEM-T4 cells. However, infectious viruses were derived from constructs in which the PBS had been altered to sequences complementary to either tRNA(1,2Lys) or tRNA(Phe), although mutated forms showed significant lags in replication efficiency in comparison with wild types. Molecular analysis of reverse-transcribed DNA in cells infected by the mutated viruses indicated that both tRNA(1,2Lys) and tRNA(Phe) could function as primers for reverse transcription during the early stages of infection. Sequencing of full-length proviral DNA, obtained 6 days after infection, revealed the mutated PBS, indicating that a complete cycle of reverse transcription had occurred. During subsequent rounds of infection, reversion of the mutated PBS to wild-type sequences was observed, accompanied by increased production of viral gene products. Reversion to wild-type PBS sequences was confirmed both by specific PCR analysis, using distinct primer pairs, and by direct sequencing of amplified segments. We also performed endogenous in vitro reverse transcription experiments in which synthesis of minus-strand strong-stop viral DNA was primed from a synthetic RNA template containing a PBS complementary to various tRNA isoacceptors. These results showed that tRNA(3Lys) was a much more efficient primer of such reactions than either tRNA(1,2Lys) or tRNA(Phe).

Proteolytic processing of the P2/nucleocapsid cleavage site is critical for human immunodeficiency virus type 1 RNA dimer maturation

Differences in virion RNA dimer stability between mature and protease-defective (immature) forms of human immunodeficiency virus type 1 (HIV-1) suggest that maturation of the viral RNA dimer is regulated by the proteolytic processing of the HIV-1 Gag and Gag-Pol precursor proteins. However, the proteolytic processing of these proteins occurs in several steps denoted primary, secondary, and tertiary cleavage events and, to date, the processing step associated with formation of stable HIV-1 RNA dimers has not been identified. We show here that a mutation in the primary cleavage site (p2/nucleocapsid [NC]) hinders formation of stable virion RNA dimers, while dimer stability is unaffected by mutations in the secondary (matrix/capsid [CA], p1/p6) or a tertiary cleavage site (CA/p2). By introducing mutations in a shared cleavage site of either Gag or Gag-Pol, we also show that the cleavage of the p2/NC site in Gag is more important for dimer formation and stability than p2/NC cleavage in Gag-Pol. Electron microscopy analysis of viral particles shows that mutations in the primary cleavage site in Gag but not in Gag-Pol inhibit viral particle maturation. We conclude that virion RNA dimer maturation is dependent on proteolytic processing of the primary cleavage site and is associated with virion core formation.

The thermal stability and structural site-distribution of Eu3+ ions in the red-emitting phosphors Ca9Eu2W4O24 and Sr9Eu2W4O24

The red-emitting phosphors Ca₉Eu₂W₄O₂₄ and Sr₉Eu₂W₄O₂₄ were synthesized by the solid-state reaction method. The crystal phases were characterized by X-ray powder diffraction. The photoluminescence excitation and emission spectra were investigated. The luminescence excitation and emission spectra confirm that the phosphors are efficiently excited by near UV light. The dependence of luminescence intensities on the heating temperatures was investigated. The Ca₉Eu₂W₄O₂₄ phosphor exhibits higher thermal stability than that of Sr₉Eu₂W₄O₂₄. The crystallographic sites for Eu³⁺ ions in Ca₉Eu₂W₄O₂₄ and Sr₉Eu₂W₄O₂₄ are investigated by the site-selective excitation spectra in the ⁵D₀ → ⁷F₀ wavelength region. It is identified that the Eu³⁺ ions occupy only M sites (statistically occupied by 0.5Eu and 0.5Ca) in Ca₉Eu₂W₄O₂₄ and, however, the Eu³⁺ ions can substitute both M sites (Eu³⁺ + Sr²⁺) and Sr²⁺ sites in Sr₉Eu₂W₄O₂₄. The luminescence spectra and the thermal stability are discussed on the basis of the crystal structure, Eu³⁺ site-distributions and the energy transfer.

Individual foraging site fidelity in lactating New Zealand fur seals : Continental shelf vs. oceanic habitats

Wide-ranging marine central place foragers often exhibit foraging site fidelity to oceanographic features over differing spatial scales (i.e., localized coastal upwellings and oceanic fronts). Few studies have tested how the degree of site fidelity to foraging areas varies in relation to the type of ocean features used. In order to determine how foraging site fidelity varied between continental shelf and oceanic foraging habitats, 31 lactating New Zealand fur seals (Arctocephalus australis forsteri1) were satellite tracked over consecutive foraging trips (14–108 d). Thirty-seven foraging trips were recorded from 11 females that foraged on the continental shelf, in a region associated with a coastal upwelling, while 65 foraging trips were recorded from 20 females that foraged in oceanic waters. There were no significant differences in the mean bearings (to maximum distance) of individual's consecutive foraging trips, suggesting individual fidelity to foraging areas. However, overlap in area and time spent in area varied considerably between continental shelf and oceanic foragers. Females that foraged on the continental shelf had significantly greater overlap in consecutive foraging trips when compared to females that foraged in oceanic waters (overlap in 5 × 5 km grid cells visited on consecutive trips 55.9%± 20.4% and 13.4%± 7.6%, respectively). Females that foraged on the continental shelf also spent significantly more time within the same grid cell than females that foraged in oceanic waters (maximum time spent in 5 × 5 km grid cells: 14%± 5% and 4%± 2%, respectively). This comparatively high foraging site fidelity may reflect the concentration of productivity associated with a coastal upwelling system, the Bonney Upwelling. Lower foraging site fidelity recorded by seals that foraged in oceanic waters implies a lower density/larger scale habitat, where prey are more dispersed or less predictable at fine scales, when compared to the continental shelf region.

Increasing the level of sustainability via off-site production : a study of the residential construction sector in China

Even though the use of off-site production in the construction process for residential buildings is seen as a viable means of increasing the level of sustainability with respects to its significant economic, environmental and social contributions, there have been very few studies to-date into off-site construction within the context of developing countries where the places have crucial demands for accommodations. Accordingly this research involves the rapidly growing developing nation, China, as a representative developing country with the objective to investigate the current status of off-site practices in the urban residential construction sector. This paper also examines the factors affecting demand for and potential barriers against a wider uptake of off-site technologies in China. The findings provide a rare insight into off-site construction in China’s housing industry and this information will be transferable to other developing countries. This study also contributes to a broader understanding about the off-site construction in developing from an international perspective.

Effects of an invasive species on refuge-site selection by native fauna : the impact of cane toads on native frogs in the Australian tropics

Invasive species can induce shifts in habitat use by native taxa: either by modifying habitat availability, or by repelling or attracting native species to the vicinity of the invader. The ongoing invasion of cane toads (Rhinella marina) through tropical Australia might affect native frogs by affecting refuge-site availability, because both frogs and toads frequently shelter by day in burrows. Our laboratory and field studies in the wet-dry tropics show that native frogs of at least three species (Litoria tornieri, Litoria nasuta and Litoria dahlii) preferentially aggregate with conspecifics, and with (some) other species of native frogs. However, the frogs rarely aggregated with cane toads either in outdoor arenas or in standardized experimental burrows that we monitored in the field. The native frogs that we tested either avoided burrows containing cane toads (or cane toad scent) or else ignored the stimulus (i.e. treated such a burrow in the same way as they did an empty burrow). Native frogs selected a highly non-random suite of burrows as diurnal retreat sites, whereas cane toads were less selective. Hence, even in the absence of toads, frogs do not use many of the burrows that are suitable for toads. The invasion of cane toads through tropical Australia is unlikely to have had a major impact on retreat-site availability for native frogs.

Natal site and offshore swimming influence fitness and long-distance ocean transport in young sea turtles

Although long-distance transport of marine organisms is constrained by numerous oceanic and biological factors, some species have evolved life-histories reliant on such movements. We examine the factors that promote long-distance transport in a transoceanic migrant, young loggerhead sea turtles (Caretta caretta), from the southeastern U.S. Empirical data from near-surface buoys and simulations in two ocean circulation models indicated that passive drifters are often retained for long periods shoreward of oceanic fronts that delineate coastal and offshore waters. Further simulations revealed that offshore swimming aided newly hatched turtles in moving past fronts and increased turtles’ probability of survival, reaching distant foraging grounds, and encountering favorable temperatures. Swimming was most beneficial in regions that were more favorable under scenarios assuming passive drift. These results have broad implications for understanding the movement processes of many marine species, highlighting likely retention of more planktonic species and potential for dispersal in more nektonic species.