pilF polymorphism-based real-time PCR to distinguish Vibrio vulnificus strains of human health relevance

The Gram-negative bacterium Vibrio vulnificus is a common inhabitant of estuarine environments. Globally, V. vulnificus is a significant foodborne pathogen capable of causing necrotizing wound infections and primary septicemia, and is a leading cause of seafood-related mortality. Unfortunately, molecular methods for the detection and enumeration of pathogenic V. vulnificus are hampered by the genetically diverse nature of this pathogen, the range of different biotypes capable of infecting humans and aquatic animals, and the fact that V. vulnificus contains pathogenic as well as non-pathogenic variants. Here we report an alternative approach utilizing the development of a real-time PCR assay for the detection of pathogenic V. vulnificus strains based on a polymorphism in pilF, a gene previously indicated to be associated with human pathogenicity. Compared to human serum reactivity, the real-time PCR assay successfully detected pathogenic strains in 46 out of 47 analysed V. vulnificus isolates (97.9%). The method is also rapid, sensitive, and more importantly can be reliably utilised on biotype 2 and 3 strains, unlike other current methods for V. vulnificus virulence differentiation

Zinc is a transmembrane agonist that induces platelet activation in a tyrosine phosphorylation-dependent manner

Following platelet adhesion and primary activation at sites of vascular injury, secondary platelet activation is induced by soluble platelet agonists, such as ADP, ATP, thrombin and thromboxane. Zinc ions are also released from platelets and damaged cells and have been shown to act as a platelet agonist. However, the mechanism of zinc-induced platelet activation is not well understood. Here we show that exogenous zinc gains access to the platelet cytosol and induces full platelet aggregation that is dependent on platelet protein tyrosine phosphorylation, PKC and integrin αIIbβ3 activity and is mediated by granule release and secondary signalling. ZnSO4 increased the binding affinity of GpVI, but not integrin α2β1. Low concentrations of ZnSO4 potentiated platelet aggregation by collagen-related peptide (CRP-XL), thrombin and adrenaline. Chelation of intracellular zinc reduced platelet aggregation induced by a number of different agonists, inhibited zinc-induced tyrosine phosphorylation and inhibited platelet activation in whole blood under physiologically relevant flow conditions. Our data are consistent with a transmembrane signalling role for zinc in platelet activation during thrombus formation

Phylogenetic Relationships in Pterodroma Petrels Are Obscured by Recent Secondary Contact and Hybridization

The classification of petrels (Pterodroma spp.) from Round Island, near Mauritius in the Indian Ocean, has confounded researchers since their discovery in 1948. In this study we investigate the relationships between Round Island petrels and their closest relatives using evidence from mitochondrial DNA sequence data and ectoparasites. Far from providing clear delimitation of species boundaries, our results reveal that hybridization among species on Round Island has led to genetic leakage between populations from different ocean basins. The most common species on the island, Pterodroma arminjoniana, appears to be hybridizing with two rarer species (P. heraldica and P. neglecta), subverting the reproductive isolation of all three and allowing gene flow. P. heraldica and P. neglecta breed sympatrically in the Pacific Ocean, where P. arminjoniana is absent, but no record of hybridization between these two exists and they remain phenotypically distinct. The breakdown of species boundaries in Round Island petrels followed environmental change (deforestation and changes in species composition due to hunting) within their overlapping ranges. Such multi-species interactions have implications not only for conservation, but also for our understanding of the processes of evolutionary diversification and speciation

Identification of a major GpVI-binding locus in human type III collagen

We have analyzed the adhesion of human and murine platelets, and of recombinant human and murine GpVI ectodomains, to synthetic triple-helical collagen-like peptides. These included 57 peptides derived from the sequence of human type III collagen and 9 peptides derived from the cyanogen bromide fragment of bovine type III collagen, α1(III)CB4. We have identified several peptides that interact with GpVI, in particular a peptide designated III-30 with the sequence GAOGLRGGAGPOGPEGGKGAAGPOGPO. Both human and murine platelets bound to peptide III-30 in a GpVI-dependent manner. III-30 also supported binding of recombinant GpVI ectodomains. Cross-linked III-30 induced aggregation of human and murine platelets, although with a lower potency than collagen-related peptide. Modifications of the peptide sequence indicated that the hydroxyproline residues play a significant role in supporting its GpVI reactivity. However, many peptides containing OGP/GPO motifs did not support adhesion to GpVI. These data indicate that the ability of a triple-helical peptide to bind GpVI is not solely determined by the presence or spatial arrangement of these OGP/GPO motifs within the peptides

Sexual size dimorphism, spatial segregation and sex-biased bycatch of southern and northern royal albatrosses in pelagic longline fisheries

Bycatch in longline fisheries is a major contributor to the global decline of albatrosses. Sexual segregation at sea often leads to unequal overlap with different fisheries, resulting in sex-biased bycatch, exacerbating the impact on a population level. In great albatrosses (Diomedea spp.), males (the larger sex) tend to spend more time at higher latitudes than females, attributed to competitive exclusion or differences in flight performance mediated by the pronounced sexual size dimorphism (SSD). Consequently, larger numbers of females are bycaught in pelagic longline fisheries in subtropical and temperate areas. Although this has been shown for Diomedea exulans, it has not been confirmed for all great albatross species. Here we examined the degree of SSD and developed discriminant functions to determine species and sex in D. epomophora and D. sanfordi; species that are often killed in several fisheries in the Southern Hemisphere. Based on a large sample of albatrosses bycaught off Uruguay, both species showed substantial SSD. Discriminant functions assigned species and sex to otherwise indeterminate individuals with 90–100% accuracy. Based on all birds identified (n=128), bycatch in the pelagic longline fishery was female-biased, indicating sexual segregation at sea. The discriminant functions presented enable species and sex to be identified, providing critical data for future bycatch assessments

The low-frequency allele of the platelet collagen signaling receptor glycoprotein VI is associated with reduced functional responses and expression

Interaction of platelets with collagen under conditions of blood flow is a multistep process with tethering via glycoprotein IbIXV (GPIbIXV) over von Willebrand factor, adhesion by direct interaction with the integrin GPIaIIa, and signaling via GPVI. GPVI can be specifically agonized by cross-linked collagen-related peptide (CRP-XL), which results in a signaling cascade very similar to that evoked by native collagen. The GPVI gene has 2 common alleles that differ by 3 replacements in the glycosylated stem and 2 in the cytoplasmic domain. We used CRP-XL to elucidate the variation in responses observed in platelet function in different individuals. We observed a 3-fold difference in the response to CRP-XL in platelet aggregation when comparing platelets from 10 high-frequency allele homozygotes with 8 low-frequency ones (2-way analysis of variance [ANOVA], P < .0001). The difference in functional responses was reflected in fibrinogen binding and in downstream signaling events as measured by tyrosine phosphorylation, the expression of P-selectin, and the binding of annexin V and the generation of thrombin on the platelet surface (2-way ANOVA, P < .001). Platelets homozygous for the low-frequency allele tended to be less able to form a thrombus on a collagen surface in flowing whole blood or in the platelet function analyzer–100 (t test, P = .065 and P = .061, respectively). The functional difference was correlated to a difference in total and membrane-expressed GPVI measured by monoclonal and polyclonal antibodies. This study demonstrates for the first time that platelet function may be altered by allelic differences in GPVI

A development of ecological risk screening with an application to fisheries off SW England

This work was funded by the Sea Fish Industry Authority.A development of the ecological risk screening (ERS) technique, scale intensity and consequence analysis (SICA), is described and application to the varied fisheries and ecosystem off the southwest of England on behalf of an industry steering group (SG) is summarized. The purpose was to prioritize ecological risks systematically and consistently in relation to policy goals agreed by the SG. Scientists listed and advised on ecosystem components, their units (individual species, stocks, habitats, or communities) and attributes, as well as agents of change in the SW, their activities, and generalized effects relevant to the policy goals. A working group (WG) of fishers, fishery observers, technical advisors, and marine scientists paired each unit with the activity thought most likely to impact the most sensitive policy goal, then scored risk according to defined rules spatially, temporally, and as intensity and duration of effects. The geometric mean of the four scores, slightly adjusted for unscored factors if necessary, was the relative impact score (RIS). With this standardized method, the main aspects of risk were considered separately and independently, thereby assisting objective prioritization. Nineteen unit–activity pairs were listed as priority risks (RIS >3) in the SW region during a 2-d meeting that fully exploited the wide range of information and experience available at the WG. Socio-economics was not considered. The ERS for the SW was designed to be compatible with other similar ERSs that might be carried out for neighbouring marine regions. ERS can minimize extra monitoring needed for ecosystem management and, in principle, collaborating non-fishery agents of change could be included. By engaging all stakeholders in the setting of initial priorities for action and by assembling all available sources of information, ERS offers a useful starting point for holistic ecosystem management.PostprintPeer reviewe