Isolation of single circulating trophoblasts from maternal circulation for noninvasive fetal copy number variant profiling

Objective

To develop a multi-step workflow for the isolation of circulating extravillous trophoblasts (cEVTs) by describing the key steps enabling a semi-automated process, including a proprietary algorithm for fetal cell origin genetic confirmation and copy number variant (CNV) detection.

Methods

Determination of the limit of detection (LoD) for submicroscopic CNV was performed by serial experiments with genomic DNA and single cells from Coriell cell line biobank with known imbalances of different sizes. A pregnancy population of 372 women was prospectively enrolled and blindly analyzed to evaluate the current workflow.

Results

An LoD of 800 Kb was demonstrated with Coriell cell lines. This level of resolution was confirmed in the clinical cohort with the identification of a pathogenic CNV of 800 Kb, also detected by chromosomal microarray. The mean number of recovered cEVTs was 3.5 cells per sample with a significant reverse linear trend between gestational age and cEVT recovery rate and number of recovered cEVTs. In twin pregnanices, evaluation of zygosity, fetal sex and copy number profiling was performed in each individual cell.

Conclusion

Our semi-automated methodology for the isolation and single-cell analysis of cEVTS supports the feasibility of a cell-based noninvasive prenatal test for fetal genomic profiling.     

The current and future management of wild mammals hunted with dogs in England and Wales

There is increasing concern about the use of lethal methods to control wild mammal populations, especially those methods that may have significant impacts on animal welfare. The continued use of dogs to hunt wild mammals in England and Wales, principally foxes (Vulpes vulpes), red deer (Cervus elaphus), brown hares (Lepus europaeus) and mink (Mustela vison), has become a focus for political debate and has been the subject of a recent UK government inquiry. This paper reports the results of a questionnaire study to quantify the use, effectiveness and acceptability of the different methods currently used to manage these four species, and future changes in management following a possible ban on hunting with dogs. There was no straightforward relationship between culling pressure and perceived pest status of the different species from the questionnaire data. For foxes and brown hares, the proportion of land managers (practitioners) carrying out lethal control was higher than that considering these species to be pests. However, the reverse was the case for mink. The most frequently used and effective control methods, which were most acceptable to practitioners and public alike, were various forms of shooting. The general public perceived hunting with dogs as one of the least acceptable means of control for all four species. Practitioners thought that hunting with dogs for red deer and the use of terriers against foxes were among the least acceptable forms of control, but considered hunting with dogs in other situations and for other species to be relatively acceptable. Most practitioners said a ban on hunting with dogs would make no difference to their management of the four species. A ban on hunting with dogs would have minimal impact on populations of foxes, red deer and mink, but it may be of conservation benefit to hares.     

Predicting Interactions among Fishing,Ocean Warming,and Ocean Acidification in a Marine System with Whole‐Ecosystem Models

An important challenge for conservation is a quantitative understanding of how multiple human stressors will interact to mitigate or exacerbate global environmental change at a community or ecosystem level. We explored the interaction effects of fishing, ocean warming, and ocean acidification over time on 60 functional groups of species in the southeastern Australian marine ecosystem. We tracked changes in relative biomass within a coupled dynamic whole‐ecosystem modeling framework that included the biophysical system, human effects, socioeconomics, and management evaluation. We estimated the individual, additive, and interactive effects on the ecosystem and for five community groups (top predators, fishes, benthic invertebrates, plankton, and primary producers). We calculated the size and direction of interaction effects with an additive null model and interpreted results as synergistic (amplified stress), additive (no additional stress), or antagonistic (reduced stress). Individually, only ocean acidification had a negative effect on total biomass. Fishing and ocean warming and ocean warming with ocean acidification had an additive effect on biomass. Adding fishing to ocean warming and ocean acidification significantly changed the direction and magnitude of the interaction effect to a synergistic response on biomass. The interaction effect depended on the response level examined (ecosystem vs. community). For communities, the size, direction, and type of interaction effect varied depending on the combination of stressors. Top predator and fish biomass had a synergistic response to the interaction of all three stressors, whereas biomass of benthic invertebrates responded antagonistically. With our approach, we were able to identify the regional effects of fishing on the size and direction of the interacting effects of ocean warming and ocean acidification. Predicción de Interacciones entre Pesca, Calentamiento de Océanos y Acidificación de Océanos en un Sistema Marino con Modelos de Ecosistemas Completos