Evidence for Changes in Estuarine Zooplankton Fostered by Increased Climate Variance

Estuaries are among the most valuable aquatic systems in terms of their services to human welfare. They offer an ideal framework to assess multiscale processes linking climate and food web dynamics through the hydrological cycle. Resolving food web responses to climate change is fundamental to resilience management of these threatened ecosystems under global change scenarios. Here, we examined the temporal variability of the plankton food web in the Mondego Estuary, central Iberian Peninsula, over the period 2003 to 2012. The results pointed out a cascading effect from climate to plankton communities that follow a non-stationary behavior shaped by the climate variance envelope. Concurrent changes in hydrographic processes at the regional, that is, upwelling intensity, and local, that is, estuarine hydrology, scales were driven by climatic forcing promoted by the North Atlantic Oscillation; the influence of which permeated the physical environment in the estuary affecting both autotrophic and heterotrophic communities. The most conspicuous change arose around 2008 and consisted of an obvious decrease in freshwater taxa along with a noticeable increase in marine organisms, mainly driven by gelatinous zooplankton. The observed increase in small-sized cosmopolitan copepods, that is, Clausocalanus arcuicornis, Oithona plumifera, thermophilic species, that is, Penilia avirostris, and gelatinous zooplankton suggests a structural change in the Mondego plankton community. These results provide empirical support to the expectation that expanding climate variance changes plankton structure and functioning, likely fostering trophic interactions in pelagic food webs.     

Complete telomere-to-telomere de novo assembly of the Plasmodium falciparum genome through long-read (>11?kb), single molecule,real-time sequencing

The application of next-generation sequencing to estimate genetic diversity of Plasmodium falciparum, the most lethal malaria parasite, has proved challenging due to the skewed AT-richness [∼80.6% (A + T)] of its genome and the lack of technology to assemble highly polymorphic subtelomeric regions that contain clonally variant, multigene virulence families (Ex: var and rifin). To address this, we performed amplification-free, single molecule, real-time sequencing of P. falciparum genomic DNA and generated reads of average length 12 kb, with 50% of the reads between 15.5 and 50 kb in length. Next, using the Hierarchical Genome Assembly Process, we assembled the P. falciparum genome de novo and successfully compiled all 14 nuclear chromosomes telomere-to-telomere. We also accurately resolved centromeres [∼90–99% (A + T)] and subtelomeric regions and identified large insertions and duplications that add extra var and rifin genes to the genome, along with smaller structural variants such as homopolymer tract expansions. Overall, we show that amplification-free, long-read sequencing combined with de novo assembly overcomes major challenges inherent to studying the P. falciparum genome. Indeed, this technology may not only identify the polymorphic and repetitive subtelomeric sequences of parasite populations from endemic areas but may also evaluate structural variation linked to virulence, drug resistance and disease transmission.     

Status of Savi's pipistrelle Hypsugo savii (Chiroptera) and range expansion in Central and south‐eastern Europe: a review

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Soil nitrogen fluctuations in citrus plots fertilized with sulfur-coated urea and ammonium nitro sulfate

Summary The release of nitrogen from a slow release fertilizer (sulfur-coated urea, SCU) in an orange orchard was studied during three annual cycles and at three soil depths. The release of N from SCU was compared with that from a standard fertilizer (ammonium nitrate sulfate, ANS). The amounts of available soil nitrogen were compared at different periods of the year and for the whole year. The SCU granules maintained higher levels of available nitrogen in the soil during critical periods. Soil N levels were similar between treatments consisting of 1500 g of N from ANS and 750 g of N from SCU.     

Effect of sex pheromone emission on the attraction of Lobesia botrana

Since the discovery of Lobesia botrana Denis & Schiffermüller (Lepidoptera: Tortricidae) sex pheromone, it has played an important role in the control and detection of this pest, for example, through the use of pheromone‐baited traps and mating disruption techniques. Rubber septa are the most common pheromone dispensers used in monitoring traps, but often dispenser performance is not optimized. The key to improve methods based on pheromones as attractants (monitoring, mass trapping, or ‘attract and kill’) is to know the optimum emission interval, because release rates can strongly affect the attraction. In this work, five levels of pheromone load with different release rates were compared in traps using mesoporous pheromone dispensers to investigate the optimum release rate maximizing L. botrana catches. Residual pheromone loads of the dispensers were extracted and quantified by gas chromatography, to study release profiles and to estimate the various emission levels. The efficacy of pheromone emission was measured in field trials as number of moths caught. A quadratic model was fitted to relate the numbers caught vs. the daily emission rates. The resulting quadratic term was statistically significant, confirming the existence of a relative maximum for L. botrana catches. Taking into account that the trial was carried out only in one location, an optimum emission value of ca. 400 μg per day could be considered to enhance the attraction of L. botrana under West‐Mediterranean weather conditions.     

Identification and genetic diversity analysis of strain Pseudomonas syringae S5: A pathogen responsible for bacteriosis in Avena sativa plants

The genus Pseudomonas includes pathogenic species P. syringae, which can be found in various agricultural environments and which can affect a wide variety of plants, causing significant economic losses when the environmental conditions for its proliferation are optimal. Comprehensive characterizations of phytopathogenic bacteria belonging to the genus Pseudomonas are scarce in Argentina. In this work, the tabtoxin‐producing strain Pseudomonas S5, isolated from oat, was identi?ed as a P. syringae through biochemical tests such as the LOPAT test, and genetic tests such as the analysis of the small subunit ribosomal RNA gene (16S rRNA) sequence and repetitive elements, using BOX and ERIC primers. It was also determined that this phytopathogen is potentially capable of infecting other crops of agricultural importance for our region, such as soybean. This ability to infect different hosts gives it an adaptive advantage that allows it to endure seasonal changes in the environment where it lives. Our work contributes to the physiological classification of the phytopathogen P. syringae S5 isolated from our region, as well as to the knowledge about its range of potential hosts.     

Prevailingly cationic agmatine-based amphoteric polyamidoamine as a nontoxic, nonhemolytic, and "stealthlike" DNA complexing agent and transfection promoter

AGMA1, a prevailingly cationic amphoteric polyamidoamine obtained by polyaddition of (4-aminobutyl)guanidine (agmatine) to 2,2-bis(acrylamido)acetic acid, was studied as a potential DNA carrier and transfection promoter. Fluorescein-labeled AGMA1 was prepared by conjugation with fluorescein isothiocyanate and its cell uptake, blood permanence, and body distribution studied. In spite of its cationic character, AGMA1 is neither toxic nor hemolytic in the pH range 4.0-7.4, circulates for a long time in the blood without preferentially localizing in the liver, easily enters HT-29 cells, gives stable complexes with DNA, and is endowed with good transfection efficiency, suggesting the ability to transport in the cytoplasm a DNA payload without any measurable membranolytic activity. If compared with other transfection promoters, including polyamidoamines of different structures, AGMA1 is apparently endowed with a unique combination of desirable requirements for a nonviral DNA polymer carrier and warrants potential as a transfection agent in vivo.     

Bimodal loop-loop interactions increase the affinity of RNA aptamers for HIV-1 RNA structures

Multiple loop-loop interactions between adjacent RNA hairpins regulate gene expression in different organisms. To demonstrate that such natural interactions could be mimicked for generating RNA ligands that are able to recognize simultaneously at least two structured RNA targets, a double kissing complex model was designed. The target consisted of two HIV-1 transactivating responsive (TAR) RNA variants, BRU and MAL, connected by a non-nucleotidic linker. The double ligand was generated by combining the corresponding hairpin aptamers, R06BRU and R06MAL, identified previously by in vitro selection [Ducongé, F., and Toulmé, J. J (1999) RNA 5, 1605-1614]. The resulting interaction was analyzed by thermal denaturation monitored by UV spectroscopy, electrophoretic mobility shift assays (EMSAs), and surface plasmon resonance (SPR) experiments. The bimodal complex was characterized by a binding equilibrium constant increased by at least 1 order of magnitude compared to that of the complexes between the individual parent hairpins. This resulted from a slower dissociation rate. We then made use of such a strategy for targeting two structured functional motifs of the folded 5' untranslated region (5'UTR) of HIV-1. Two bivalent RNA ligands were designed that targeted simultaneously the TAR and dimerization initiation site (DIS) hairpins or the TAR and poly(A) ones. The results show that these ligands also displayed enhanced affinity for their target compared to the individual molecules. The work reported here suggests that bimodal structured RNA ligands might provide a way of increasing the affinity of aptamers for folded RNA targets.