Small home ranges and high site fidelity in red knots (Calidris c. canutus) wintering on the Banc d’Arguin, Mauritania

Using automated and manual radio-telemetry and resightings of individual colour-ringed birds, we assessed the daily use of space of red knots Calidris canutus canutus at a tropical wintering area along the Sahara coast, the Banc d Arguin in Mauritania. Confirming earlier suggestions, we found that birds were very faithful to their roosts and that the daily foraging range was small; in the course of several winter months birds used an area of only 2 16 km2 of intertidal area. We found no differences between their movements in daylight and at night. Additionally, individuals seem to return to exactly the same locations in subsequent winters. This pattern is very different from red knots wintering in the temperate Wadden Sea. Here, they readily change roost sites and easily cover areas of about 800 km2 in the course of weeks but, just as in Mauritania, no differences between day and night are apparent. In northern Patagonia and north-western Australia, red knots have range sizes closer to those on the Banc d Arguin, but here they do show differences in space use between day and night. Ecological explanations for these contrasting patterns require further comparative data based on in-depth studies on the predictability of the food base and the presence of diurnal and nocturnal predators.

Densities of individually marked migrants away from the marking site to estimate population sizes : a test with three wader populations

Capsule Population estimates based on the mark–resighting method can be a useful alternative to population-wide counts.

Aims To investigate whether the mark–resighting method can be used as an alternative to counts to estimate the size of wader populations.

Methods Individual colour-marking and subsequent resightings allowed accurate estimates of annual survival for three populations of waders, on which basis we could estimate the actual number of marked birds alive. Densities of marked birds were determined on sites away (2000–4300 km) from the ringing locations expecting marked birds to be randomly distributed among non-marked conspecifics. Population sizes are estimated by combining these densities with the number of marked birds alive.

Results We found indications that the distribution of marked birds was indeed random in the locations away from the site of marking. The estimated population size of Red Knot Calidris canutus canutus was in accordance with the most recent estimates based on counts. Our estimate of the Calidris c. islandica population was somewhat lower, and that of the Bar-tailed Godwit Limosa lapponica taymyrensis population was considerably lower than the latest estimates based on counts.

Conclusion Population estimates based on the mark–resighting method can be a useful alternative for, or addition to, population-wide counts, as long as the assumption of random distribution of marked birds at the reading sites is taken into account. We conclude that the Afro-Siberian Bar-tailed Godwit population has recently decreased in size or has been substantially overestimated during the counts.

Yhh1p/Cft1p directly links poly(A) site recognition and RNA polymerase II transcription termination

RNA polymerase II (pol II) transcription termination requires co-transcriptional recognition of a functional polyadenylation signal, but the molecular mechanisms that transduce this signal to pol II remain unclear. We show that Yhh1p/Cft1p, the yeast homologue of the mammalian AAUAAA interacting protein CPSF 160, is an RNA-binding protein and provide evidence that it participates in poly(A) site recognition. Interestingly, RNA binding is mediated by a central domain composed of predicted -propeller-forming repeats, which occurs in proteins of diverse cellular functions. We also found that Yhh1p/Cft1p bound specifically to the phosphorylated C-terminal domain (CTD) of pol II in vitro and in a two-hybrid test in vivo. Furthermore, transcriptional run-on analysis demonstrated that yhh1 mutants were defective in transcription termination, suggesting that Yhh1p/Cft1p functions in the coupling of transcription and 3'-end formation. We propose that direct interactions of Yhh1p/Cft1p with both the RNA transcript and the CTD are required to communicate poly(A) site recognition to elongating pol II to initiate transcription termination.

Phenylalanine-544 plays a key role in substrate and inhibitor binding by providing a hydrophobic packing point at the active site of insulin-regulated aminopeptidase

Inhibitors of insulin-regulated aminopeptidase (IRAP) improve memory and are being developed as a novel treatment for memory loss. In this study, the binding of a class of these inhibitors to human IRAP was investigated using molecular docking and site-directed mutagenesis. Four benzopyran-based IRAP inhibitors with different affinities were docked into a homology model of the catalytic site of IRAP. Two 4-pyridinyl derivatives orient with the benzopyran oxygen interacting with the Zn2+ ion and a direct parallel ring-stack interaction between the benzopyran rings and Phe544. In contrast, the two 4-quinolinyl derivatives orient in a different manner, interacting with the Zn2+ ion via the quinoline nitrogen, and Phe544 contributes an edge-face hydrophobic stacking point with the benzopyran moiety. Mutagenic replacement of Phe544 with alanine, isoleucine, or valine resulted in either complete loss of catalytic activity or altered hydrolysis velocity that was substrate-dependent. Phe544 is also important for inhibitor binding, because these mutations altered the Ki in some cases, and docking of the inhibitors into the corresponding Phe544 mutant models revealed how the interaction might be disturbed. These findings demonstrate a key role of Phe544 in the binding of the benzopyran IRAP inhibitors and for optimal positioning of enzyme substrates during catalysis.

A Ramsar wetland in crisis the Coorong, Lower Lakes and Murray Mouth, Australia

The state of global freshwater ecosystems is increasingly parlous with water resource development degrading high-conservation wetlands. Rehabilitation is challenging because necessary increases in environmental flows have concomitant social impacts, complicated because many rivers flow between jurisdictions or countries. Australia's MurrayDarling Basin is a large river basin with such problems encapsulated in the crisis of its Ramsar-listed terminal wetland, the Coorong, Lower Lakes and Murray Mouth. Prolonged drought and upstream diversion of water dropped water levels in the Lakes below sea level (20092010), exposing hazardous acid sulfate soils. Salinities increased dramatically (e.g. South Lagoon of Coorong>200gL-1, cf. modelled natural 80gL-1), reducing populations of waterbirds, fish, macroinvertebrates and littoral plants. Calcareous masses of estuarine tubeworms (Ficopomatus enigmaticus) killed freshwater turtles (Chelidae) and other fauna. Management primarily focussed on treating symptoms (e.g. acidification), rather than reduced flows, at considerable expense (≥AU$2 billion). We modelled a scenario that increased annual flows during low-flow periods from current levels up to one-third of what the natural flow would have been, potentially delivering substantial environmental benefits and avoiding future crises. Realisation of this outcome depends on increasing environmental flows and implementing sophisticated river management during dry periods, both highly contentious options.

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).