Matching productivity to resource availability in a small predator, the stoat (Mustela erminea)

Stoats (Mustela erminea), introduced to New Zealand in the late nineteenth century, are common in New Zealand beech (Nothofagus sp.) forests, where populations of feral house mice (Mus musculus) fluctuate between years much as voles do in the northern hemisphere. We present new field evidence and two models demonstrating (i) a strong correlation between density indices for young stoats in summer and for mice in the previous spring, and (ii) a significant linear relationship between productivity per female and spring density of mice up to 25 mice captures per 100 trap-nights. These models confirm that short-lived small mustelid predators dependent on fluctuating populations of prey have evolved means of matching their productivity to the prospects of success across a wide range, from total failure in rodent crash years to >12 independent young per female in rodent peak years. We suggest that the enhanced reproductive success of female stoats when rodents are abundant is due to a combination of critical improvements in both the reproductive physiology and the foraging behaviour of female stoats in rodent peak years. Conversely, a drastic shortage of rodents increases the mortality of embryos and nestlings, while the adult females are able to survive, and even remain relatively fat, on other foods

Cancer detection in human tissue samples using a fiber-tip pH probe

Abstract not availableErik P. Schartner, Matthew R. Henderson, Malcolm Purdey, Deepak Dhatrak, Tanya M. Monro, P. Grantley Gill, and David F. Calle

An organic fluorophore-nanodiamond hybrid sensor for photostable imaging and orthogonal, on-demand biosensing

Organic fluorescent probes are widely used to detect key biomolecules; however, they often lack the photostability required for extended intracellular imaging. Here we report a new hybrid nanomaterial (peroxynanosensor, PNS), consisting of an organic fluorescent probe bound to a nanodiamond, that overcomes this limitation to allow concurrent and extended cell-based imaging of the nanodiamond and ratiometric detection of hydrogen peroxide. Far-red fluorescence of the nanodiamond offers continuous monitoring without photobleaching, while the green fluorescence of the organic fluorescent probe attached to the nanodiamond surface detects hydrogen peroxide on demand. PNS detects basal production of hydrogen peroxide within M1 polarised macrophages and does not affect macrophage growth during prolonged co-incubation. This nanosensor can be used for extended bio-imaging not previously possible with an organic fluorescent probe, and is spectrally compatible with both Hoechst 33342 and MitoTracker Orange stains for hyperspectral imaging.Malcolm S. Purdey, Patrick K. Capon, Benjamin J. Pullen, Philipp Reineck, Nisha Schwarz, Peter J. Psaltis, Stephen J. Nicholls, Brant C. Gibson and Andrew D. Abel

Manganese Enhances Prion Protein Survival in Model Soils and Increases Prion Infectivity to Cells

Prion diseases are considered to be transmissible. The existence of sporadic forms of prion diseases such as scrapie implies an environmental source for the infectious agent. This would suggest that under certain conditions the prion protein, the accepted agent of transmission, can survive in the environment. We have developed a novel technique to extract the prion protein from soil matrices. Previous studies have suggested that environmental manganese is a possible risk factor for prion diseases. We have shown that exposure to manganese is a soil matrix causes a dramatic increase in prion protein survival (∼10 fold) over a two year period. We have also shown that manganese increases infectivity of mouse passaged scrapie to culture cells by 2 logs. These results clearly verify that manganese is a risk factor for both the survival of the infectious agent in the environment and its transmissibility

Hyperspectral microscopy can detect metabolic heterogeneity within bovine post-compaction embryos incubated under two oxygen concentrations (7% versus 20%)

Study Question: Can we separate embryos cultured under either 7% or 20% oxygen atmospheres by measuring their metabolic heterogeneity? Summary Answer: Metabolic heterogeneity and changes in metabolic profiles in morula exposed to two different oxygen concentrations were not detectable using traditional fluorophore and two-channel autofluorescence but were detectable using hyperspectral microscopy. What is Known Already: Increased genetic and morphological blastomere heterogeneity is associated with compromised developmental competence of embryos and currently forms the basis for embryo scoring within the clinic. However, there remains uncertainty over the accuracy of current techniques, such as PGS and time-lapse microscopy, to predict subsequent pregnancy establishment. Study Design, Size, Duration: The impact of two oxygen concentrations (7% = optimal and 20% = stressed) during post-fertilisation embryo culture was assessed. Cattle embryos were exposed to the different oxygen concentrations for 8 days (D8; embryo developmental competence) or 5 days (D5; metabolism measurements). Between 3 and 4 experimental replicates were performed, with 40–50 embryos per replicate used for the developmental competency experiment, 10–20 embryos per replicate for the fluorophore and two-channel autofluorescence experiments and a total of 21–22 embryos used for the hyperspectral microscopy study. Participants/Materials, Setting, Methods: In-vitro produced (IVP) cattle embryos were utilised for this study. Post-fertilisation, embryos were exposed to 7% or 20% oxygen. To determine impact of oxygen concentrations on embryo viability, blastocyst development was assessed on D8. On D5, metabolic heterogeneity was assessed in morula (on-time) embryos using fluorophores probes (active mitochondria, hydrogen peroxide and reduced glutathione), two-channel autofluorescence (FAD and NAD(P)H) and 18-channel hyperspectral microscopy. Main Results and the Role of Chance: Exposure to 20% oxygen following fertilisation significantly reduced total blastocyst, expanded and hatched blastocyst rates by 1.4-, 1.9- and 2.8-fold, respectively, compared to 7% oxygen (P 0.05). While there were no significant differences in two-channel autofluorescent profiles of morula exposed to 7% and 20% oxygen (main effect, P > 0.05), morula that subsequently progressed to the blastocyst stage had significantly higher levels of FAD and NAD(P)H fluorescence compared to arrested morula (P < 0.05), with no change in the redox ratio. Hyperspectral autofluorescence imaging (in 18-spectral channels) of the D5 morula revealed highly significant differences in four features of the metabolic profiles of morula exposed to the two different oxygen concentrations (P < 0.001). These four features were weighted and their linear combination revealed clear discrimination between the two treatment groups. Limitations, Reasons for Caution: Metabolic profiles were assessed at a single time point (morula), and as such further investigation is required to determine if differences in hyperspectral signatures can be detected in pre-compaction embryos and oocytes, using both cattle and subsequently human models. Furthermore, embryo transfers should be performed to determine the relationship between metabolic profiles and pregnancy success. Wider Implications of the Findings: Advanced autofluorescence imaging techniques, such as hyperspectral microscopy, may provide clinics with additional tools to improve the assessment of embryos prior to transfer.Melanie L. Sutton-McDowall, Martin Gosnell, Ayad G. Anwer, Melissa White, Malcolm Purdey, Andrew D. Abell, Ewa M. Goldys, and Jeremy G. Thompso

Localised hydrogen peroxide sensing for reproductive health

Session 10 - Chemical Sensors and Biosensors IIThe production of reactive oxygen species (ROS) is known to affect the developmental competence of embryos. Hydrogen peroxide (H₂O₂) an important reactive oxygen species, is also known to causes DNA damage and defective sperm function. Current techniques require incubating a developing embryo with an organic fluorophore which is potentially hazardous for the embryo. What we need is a localised ROS sensor which does not require fluorophores in solution and hence will allow continuous monitoring of H₂O₂ production without adversely affect the development of the embryo. Here we report studies on such a fibre-based sensor for the detection of H₂O₂ that uses a surface-bound aryl boronate fluorophore carboxyperoxyfluor-1(CPF1). Optical fibres present a unique platform due to desirable characteristics as dip sensors in biological solutions. Attempts to functionalise the fibre tips using polyelectrolyte layers and (3-aminopropyl)triethoxysilane (APTES) coatings resulted in a limited signal and poor fluorescent response to H₂O₂ due to a low tip surface density of the fluorophore. To increase the surface density, CPF1 was integrated into a polymer matrix formed on the fibre tip by a UV-catalysed polymerisation process of acrylamide onto a methacrylate silane layer. The polyacrylamide containing CPF1 gave a much higher surface density than previous surface attachment methods and the sensor was found to effectively detect H₂O₂. Using this method, biologically relevant concentrations of H₂O₂ were detected, enabling remote sensing studies into ROS releases from embryos throughout early development.Malcolm S Purdey, Erik P Schartner, Melanie L Sutton-McDowall, Lesley J Ritter, Jeremy G Thompson, Tanya M Monro, and Andrew D Abel

High precision pH measurements in biological environments using a portable optical fibre pH sensor

Abstract not availableGeorgina M. Sylvia, Erik P. Schartner, Hanna J. McLennan, Avishkar Saini, Kylie R. Dunning, Malcolm S. Purdey, Andrew D. Abell, Jeremy G. Thompso

Boronate probes for the detection of hydrogen peroxide release from human spermatozoa

Human spermatozoa are compromised by production of reactive oxygen species (ROS) and detection of ROS in spermatozoa is important for the diagnosis of male infertility. Probes 2',7'-dichlorohydrofluorescein diacetate (DCFH), dihydroethidium (DHE) and MitoSOX red (MSR) are commonly used for detecting ROS by flow cytometry, however these probes lack sensitivity to hydrogen peroxide (H2O2), which is particularly damaging to mammalian sperm cells. This study reports the synthesis and use of three aryl boronate probes, peroxyfluor-1 (PF1), carboxy peroxyfluor-1 (CPF1) and a novel probe 2(2-ethoxyethoxy)ethoxy peroxyfluor-1 (EEPF1) in human spermatozoa. PF1 and EEPF1 were found to be effective in detecting H2O2 and peroxynitrite (ONOO(-)) produced by spermatozoa when stimulated with menadione or 4-hydroxynonenal. EEPF1 was more effective at detection of ROS in spermatozoa than DCFH, DHE and MSR; furthermore it distinguished poorly motile sperm as shown by greater ROS production. EEPF1 should therefore have a significant role in diagnosis of oxidative stress in male infertility, cryopreservation, age, lifestyle and exposure to environmental toxicants.Malcolm S. Purdey, Haley S. Connaughton, Sara Whiting, Erik P. Schartner, Tanya M. Monro, Jeremy G. Thompson, R. John Aitken, Andrew D. Abel