Assessment of the range of attraction of pheromone traps to Agriotes lineatus and Agriotes obscurus

• 1 The range of attraction of YATLOR pheromone traps was studied to gain information on the number of traps needed for mass trapping of males of two Agriotes species.
• 2 Male click beetles of the species Agriotes lineatus (L.) and Agriotes obscurus (L.) (25–30 individuals per release point) were marked and released at a distance of 2, 5, 10, 15, 20 and 60 m from a pheromone trap both along and opposite to the known prevailing wind direction. Traps were regularly inspected over approximately 1 month. The percentage of recaptured beetles was calculated and analyzed using analysis of variance. Maximum sampling ranges and effective sampling areas were calculated.
• 3 Averaged over all five trials and distances, approximately 40% of the released beetles (A. lineatus and A. obscurus) were recaptured. The percentage recapture of male adults was significantly affected by release distance, whereas no differences were found for species and release direction.
• 4 Males were recaptured from all release points and the percentage recapture decreased (in part significantly) with increasing distance from 76% (2 m) to 35% (15 m) and 9% (60 m), respectively. Most of the beetles were recaptured within the first 3 days after release, independent of the distance, except 60 m. The effective sampling area for A. lineatus was 1089 m² after 12 days and increased to 1735 m² after 30 days. Corresponding values for A. obscurus were considerably higher: 1518 m² for 12 days and 2633 m² for 30 days.
• 5 We conclude that the range of attraction of the pheromone traps for A. lineatus and A. obscurus is comparatively low, providing high percentage recapture only for release distances up to 10 m. Accordingly, any approach targeted on preventing mating by male mass trapping would require a dense network of pheromone traps.

     

PCR-based species identification of Agriotes larvae

Click beetle larvae within the genus Agriotes (Coleoptera: Elateridae), commonly known as wireworms, are abundant ground-dwelling herbivores which can inflict considerable damage to field crops. In Central Europe up to 20 species, which differ in their distribution, ecology and pest status, occur in arable land. However, the identification of these larvae based on morphological characters is difficult or impossible. This hampers progress towards controlling these pests. Here, we present a polymerase chain reaction (PCR)-based approach to identify, for the first time, 17 Agriotes species typically found in Central Europe. Diagnostic sequence information was generated and submitted to GenBank, allowing the identification of these species via DNA barcoding. Moreover, multiplex PCR assays were developed to identify the nine most abundant species rapidly within a single-step reaction: Agriotes brevis, A. litigiosus, A. obscurus, A. rufipalpis, A. sordidus, A. sputator, A. ustulatus, A. lineatus and A. proximus. The latter two species remain molecularly indistinguishable, questioning their species status. The multiplex PCR assays proved to be highly specific against non-agrioted elaterid beetles and other non-target soil invertebrates. By testing the molecular identification system with over 900 field-collected larvae, our protocol proved to be a reliable, cheap and quick method to routinely identify Central European Agriotes species.     

C-terminal WxL domain mediates cell wall binding in Enterococcus faecalis and other gram-positive bacteria

Analysis of the genome sequence of Enterococcus faecalis clinical isolate V583 revealed novel genes encoding surface proteins. Twenty-seven of these proteins, annotated as having unknown functions, possess a putative N-terminal signal peptide and a conserved C-terminal region characterized by a novel conserved domain designated WxL. Proteins having similar characteristics were also detected in other low-G+C-content gram-positive bacteria. We hypothesized that the WxL region might be a determinant of bacterial cell location. This hypothesis was tested by generating protein fusions between the C-terminal regions of two WxL proteins in E. faecalis and a nuclease reporter protein. We demonstrated that the C-terminal regions of both proteins conferred a cell surface localization to the reporter fusions in E. faecalis. This localization was eliminated by introducing specific deletions into the domains. Interestingly, exogenously added protein fusions displayed binding to whole cells of various gram-positive bacteria. We also showed that the peptidoglycan was a binding ligand for WxL domain attachment to the cell surface and that neither proteins nor carbohydrates were necessary for binding. Based on our findings, we propose that the WxL region is a novel cell wall binding domain in E. faecalis and other gram-positive bacteria.     

Studying the Cu binding sites in the PrP N-terminal region: a test case for ab initio simulations

First principle ab initio molecular dynamics simulations of the Car-Parrinello type have proved to be of invaluable help in understanding the microscopic mechanisms of chemical bonding both in solid state physics and in structural biophysics. In this work we present as a test case a study of the Cu coordination mode at the Prion Protein binding sites localized in the N-terminal octarepeat region. Using medium size PC-clusters, we are able to deal with systems with up to about 350 atoms and 10(3) electrons for as long as approximately 2 ps. With a foreseeable forthcoming scaling up of the available CPU times by a factor 10(3), one can hope to be soon able to simulate systems of biological interest of realistic size and for physical times of the order of the nanosecond.     

Vesicle Associated Membrane Protein 8 (VAMP8)-mediated Zymogen Granule Exocytosis Is Dependent on Endosomal Trafficking via the Constitutive-Like Secretory Pathway

Acinar cell zymogen granules (ZG) express 2 isoforms of the vesicle-associated membrane protein family (VAMP2 and -8) thought to regulate exocytosis. Expression of tetanus toxin to cleave VAMP2 in VAMP8 knock-out (−/−) acini confirmed that VAMP2 and -8 are the primary VAMPs for regulated exocytosis, each contributing ∼50% of the response. Analysis of VAMP8^−/− acini indicated that although stimulated secretion was significantly reduced, a compensatory increase in constitutive secretion maintained total secretion equivalent to wild type (WT). Using a perifusion system to follow secretion over time revealed VAMP2 mediates an early rapid phase peaking and falling within 2–3 min, whereas VAMP8 controls a second prolonged phase that peaks at 4 min and slowly declines over 20 min to support the protracted secretory response. VAMP8^−/− acini show increased expression of the endosomal proteins Ti-VAMP7 (2-fold) and Rab11a (4-fold) and their redistribution from endosomes to ZGs. Expression of GDP-trapped Rab11a-S25N inhibited secretion exclusively from the VAMP8 but not the VAMP2 pathway. VAMP8^−/− acini also showed a >90% decrease in the early endosomal proteins Rab5/D52/EEA1, which control anterograde trafficking in the constitutive-like secretory pathway. In WT acini, short term (14–16 h) culture also results in a >90% decrease in Rab5/D52/EEA1 and a complete loss of the VAMP8 pathway, whereas VAMP2-secretion remains intact. Remarkably, rescue of Rab5/D52/EEA1 expression restored the VAMP8 pathway. Expressed D52 shows extensive colocalization with Rab11a and VAMP8 and partially copurifies with ZG fractions. These results indicate that robust trafficking within the constitutive-like secretory pathway is required for VAMP8- but not VAMP2-mediated ZG exocytosis.     

Environmental factors modulating the abundance and distribution of Callinectes danae (Decapoda: Portunidae) from two areas of the southeastern coast of Brazil

The distribution and abundance of marine organisms are directly or indirectly a function of biotic and abiotic features such as sediment and water properties. In this study we analyzed the spatial and temporal distribution of Callinectes danae in two distinct littoral areas of the northern coast of the State of São Paulo, Brazil and associated them with local geomorphologic features and environmental factors. We collected specimens on a monthly basis, from July 2001 to June 2003, using a fishing boat equipped with double-rig net, in two locations: Ubatuba (UBA) and Caraguatatuba (CAR). Collecting was carried out in seven transects with different depths, which were further subdivided into two regions: bay region (5, 10, 15 and 20 m deep); and inner shelf region (25, 30 and 35 m deep). Samples from different periods and transects differed significantly in abundance. A positive correlation was found between the abundance of C. danae and the following parameters: temperature at the bottom, organic matter content and Phi (grain diameter). By contrast, a negative correlation was established between specimen abundance and salinity at the bottom. The total abundance was highest in 5 m deep transects in both regions. Summer was the season when individuals were least abundant in both areas. We concluded that the spatial and temporal distribution of C. danae are determined by a set of environmental factors and not just by one single factor.