title: Evaluating detection limits of next-generation sequencing for the surveillance and monitoring of international marine pests

authors: Pochon, Xavier

abstract: Most surveillance programmes for marine invasive species (MIS) require considerable taxonomic expertise, are laborious, and are unable to identify species at larval or juvenile stages. Therefore, marine pests may go undetected at the initial stages of incursions when population densities are low. In this study, we evaluated the ability of the benchtop GS Junior™ 454 pyrosequencing system to detect the presence of MIS in complex sample matrices. An initial in-silico evaluation of the mitochondrial cytochrome c oxidase subunit I (COI) and the nuclear small subunit ribosomal DNA (SSU) genes, found that multiple primer sets (targeting a ca. 400 base pair region) would be required to obtain species level identification within the COI gene. In contrast a single universal primer set was designed to target the V1–V3 region of SSU, allowing simultaneous PCR amplification of a wide taxonomic range of MIS. To evaluate the limits of detection of this method, artificial contrived communities (10 species from 5 taxonomic groups) were created using varying concentrations of known DNA samples and PCR products. Environmental samples (water and sediment) spiked with one or five 160 hr old Asterias amurensis larvae were also examined. Pyrosequencing was able to recover DNA/PCR products of individual species present at greater than 0.64% abundance from all tested contrived communities. Additionally, single A. amurensis larvae were detected from both water and sediment samples despite the co-occurrence of a large array of environmental eukaryotes, indicating an equivalent sensitivity to quantitative PCR. NGS technology has tremendous potential for the early detection of marine invasive species worldwide.

text: Evaluating detection limits of next-generation sequencing for the surveillance and monitoring of international marine pests Most surveillance programmes for marine invasive species (MIS) require considerable taxonomic expertise, are laborious, and are unable to identify species at larval or juvenile stages. Therefore, marine pests may go undetected at the initial stages of incursions when population densities are low. In this study, we evaluated the ability of the benchtop GS Junior™ 454 pyrosequencing system to detect the presence of MIS in complex sample matrices. An initial in-silico evaluation of the mitochondrial cytochrome c oxidase subunit I (COI) and the nuclear small subunit ribosomal DNA (SSU) genes, found that multiple primer sets (targeting a ca. 400 base pair region) would be required to obtain species level identification within the COI gene. In contrast a single universal primer set was designed to target the V1–V3 region of SSU, allowing simultaneous PCR amplification of a wide taxonomic range of MIS. To evaluate the limits of detection of this method, artificial contrived communities (10 species from 5 taxonomic groups) were created using varying concentrations of known DNA samples and PCR products. Environmental samples (water and sediment) spiked with one or five 160 hr old Asterias amurensis larvae were also examined. Pyrosequencing was able to recover DNA/PCR products of individual species present at greater than 0.64% abundance from all tested contrived communities. Additionally, single A. amurensis larvae were detected from both water and sediment samples despite the co-occurrence of a large array of environmental eukaryotes, indicating an equivalent sensitivity to quantitative PCR. NGS technology has tremendous potential for the early detection of marine invasive species worldwide.

year: 2013

title: Tensile Properties and Fracture Behaviour of Induction Sintered Ti and Ti-6Al-4V (wt %) Powder Compacts

authors: Raynova, Stiliana (Stella) Rousseva

subjects: Titanium alloys

abstract: Induction heating of powder compacts could be a very effective method for metal powder consolidation to get the final product or as an intermediate consolidation step to produce feedstock for metal powder forging or extrusion. Our study has proven that only a few minutes of induction heating, of Ti and Ti-6Al-4V (wt %) powder compacts, increases their density dramatically and causes a significant sintering effect, as evidenced by the formation of interparticle diffusion bonding and reflected by the tensile properties of the induction sintered powder compacts. This paper presents and discusses the results of a study on the tensile properties and fracture behaviour of the Ti and Ti-6Al-4V powder compacts sintered under a variety of induction heating conditions. Keywords: Titanium alloys, powder consolidation, powder compact sintering, induction heating.

text: Tensile Properties and Fracture Behaviour of Induction Sintered Ti and Ti-6Al-4V (wt %) Powder Compacts Induction heating of powder compacts could be a very effective method for metal powder consolidation to get the final product or as an intermediate consolidation step to produce feedstock for metal powder forging or extrusion. Our study has proven that only a few minutes of induction heating, of Ti and Ti-6Al-4V (wt %) powder compacts, increases their density dramatically and causes a significant sintering effect, as evidenced by the formation of interparticle diffusion bonding and reflected by the tensile properties of the induction sintered powder compacts. This paper presents and discusses the results of a study on the tensile properties and fracture behaviour of the Ti and Ti-6Al-4V powder compacts sintered under a variety of induction heating conditions. Keywords: Titanium alloys, powder consolidation, powder compact sintering, induction heating.

year: 2011

title: Supervision and culture: Meetings at thresholds

authors: Crocket, Kathie

subjects: supervision

abstract: Counsellors are required to engage in supervision in order to reflect on, reflexively review, and extend their practice. Supervision, then, might be understood as a partnership in which the focus of practitioners and supervisors is on ethical and effective practice with all clients. In Aotearoa/New Zealand, there has recently been interest in the implications for supervision of cultural difference, particularly in terms of the Treaty of Waitangi as a practice metaphor, and when non-Māori practitioners counsel Māori clients. This article offers an account of a qualitative investigation by a group of counsellors/supervisors into their experiences of supervision as cultural partnership. Based on interviews and then using writing-as-research, the article explores the playing out of supervision’s contribution to practitioners’ effective and ethical practice in the context of Aotearoa/New Zealand, showing a range of possible accounts and strategies and discussing their effects. Employing the metaphor of threshold, the article includes a series of reflections and considerations for supervision practice when attention is drawn to difference.

text: Supervision and culture: Meetings at thresholds Counsellors are required to engage in supervision in order to reflect on, reflexively review, and extend their practice. Supervision, then, might be understood as a partnership in which the focus of practitioners and supervisors is on ethical and effective practice with all clients. In Aotearoa/New Zealand, there has recently been interest in the implications for supervision of cultural difference, particularly in terms of the Treaty of Waitangi as a practice metaphor, and when non-Māori practitioners counsel Māori clients. This article offers an account of a qualitative investigation by a group of counsellors/supervisors into their experiences of supervision as cultural partnership. Based on interviews and then using writing-as-research, the article explores the playing out of supervision’s contribution to practitioners’ effective and ethical practice in the context of Aotearoa/New Zealand, showing a range of possible accounts and strategies and discussing their effects. Employing the metaphor of threshold, the article includes a series of reflections and considerations for supervision practice when attention is drawn to difference.

year: 2013