Easy and accessible way to calibrate a fluorescence microscope and to create a microplastic identification key

We present here a technique for setting up detection limits on any fluorescent microscope in conjunction with the fluorophore Nile Red for microplastic identification. Our method also describes a rigorous morphology-specific identification key for microplastics to reduce subjectivity between researchers. The detection limits were established for nine common polymer types and five natural substrates which could result in false-positive signals when using Nile Red for microplastic identification. This method was then applied to real freshwater samples and identified particles were validated with micro-FTIR or Raman spectroscopy. This approach may reduce subjectivity in microplastic identification and counting and enhances transparency, repeatability and harmonization within microplastic research community.• Instructions for calibration of detection limits for microplastics on fluorescence microscope systems described•Microplastic identification key developed and tested to reduce false positive detection•Lower subjectivity for microplastic identification obtained using the detection limits & identification ke

The role of discharge variability in determining alluvial stratigraphy

We illustrate the potential for using physics-based modeling to link alluvial stratigraphy to large river morphology and dynamics. Model simulations, validated using ground penetrating radar data from the Río Paraná, Argentina, demonstrate a strong relationship between bar-scale set thickness and channel depth, which applies across a wide range of river patterns and bar types. We show that hydrologic regime, indexed by discharge variability and flood duration, exerts a first-order influence on morphodynamics and hence bar set thickness, and that planform morphology alone may be a misleading variable for interpreting deposits. Indeed, our results illustrate that rivers evolving under contrasting hydrologic regimes may have very similar morphology, yet be characterized by marked differences in stratigraphy. This realization represents an important limitation on the application of established theory that links river topography to alluvial deposits, and highlights the need to obtain field evidence of discharge variability when developing paleoenvironmental reconstructions. Model simulations demonstrate the potential for deriving such evidence using metrics of paleocurrent variance

Influence of Dunes on Channel‐Scale Flow and Sediment Transport in a Sand Bed Braided River

This is the final version. Available on open access from the American Geophysical Union via the DOI in this recordData availability: Project data is stored in, and available from, the UK Centre for Ecology & Hydrology (http://eidc.ceh.ac.uk).Current understanding of the role that dunes play in controlling bar and channel-scale processes and river morphodynamics is incomplete. We present results from a combined numerical modeling and field monitoring study that isolates the impact of dunes on depth-averaged and near-bed flow structure, with implications for morphodynamic modeling. Numerical simulations were conducted using the three-dimensional Computational Fluid Dynamics code OpenFOAM to quantify the time-averaged flow structure within a 400 m x 100 m channel using DEMs for which: (i) dunes and bars were present within the model; and (ii) only bar43 scale topographic features were resolved (dunes were removed). Comparison of these two simulations shows that dunes enhance lateral flows and reduce velocities over bar tops by as much as 30%. Dunes influence the direction of modeled sediment transport at spatial scales larger than individual bedforms due to their effect on topographic steering of the near-bed flow structure. We show that dunes can amplify, dampen or even reverse the deflection of sediment down lateral bar slopes, and this is closely associated with 3D and obliquely orientated dunes. Sediment transport patterns calculated using theory implemented in depth-averaged morphodynamic models suggests that gravitational deflection of sediment is still controlled by bar-scale topography, even in the presence of dunes. However, improved parameterizations of flow and sediment transport in depth-averaged morphodynamic models are needed that account for the effects of both dune- and bar- scale morphology on near-bed flow and sediment transport.Natural Environment Research Council (NERC

Quantification of bedform dynamics and bedload sediment flux in sandy braided rivers from airborne and satellite imagery

Images from specially‐commissioned aeroplane sorties (manned aerial vehicle, MAV), repeat unmanned aerial vehicle (UAV) surveys, and Planet CubeSat satellites are used to quantify dune and bar dynamics in the sandy braided South Saskatchewan River, Canada. Structure‐from‐Motion (SfM) techniques and application of a depth‐brightness model are used to produce a series of Digital Surface Models (DSMs) at low and near‐bankfull flows. A number of technical and image processing challenges are described that arise from the application of SfM in dry and submerged environments. A model for best practice is presented and analysis suggests a depth‐brightness model approach can represent the different scales of bedforms present in sandy braided rivers with low‐turbidity and shallow (< 2 m deep) water. The aerial imagery is used to quantify the spatial distribution of unit bar and dune migration rate in an 18 km reach and three ~1 km long reaches respectively. Dune and unit bar migration rates are highly variable in response to local variations in planform morphology. Sediment transport rates for dunes and unit bars, obtained by integrating migration rates (from UAV) with the volume of sediment moved (from DSMs using MAV imagery) show near‐equivalence in sediment flux. Hence, reach‐based sediment transport rate estimates can be derived from unit bar data alone. Moreover, it is shown that reasonable estimates of sediment transport rate can be made using just unit bar migration rates as measured from 2D imagery, including from satellite images, so long as informed assumptions are made regarding average bar shape and height. With recent availability of frequent, repeat satellite imagery, and the ease of undertaking repeat MAV and UAV surveys, for the first time, it may be possible to provide global estimates of bedload sediment flux for large or inaccessible low‐turbidity rivers that currently have sparse information on bedload sediment transport rates

The planform mobility of river channel confluences: Insights from analysis of remotely sensed imagery

River channel confluences are widely acknowledged as important geomorphological nodes that control the downstream routing of water and sediment, and which are locations for the preservation of thick fluvial deposits overlying a basal scour. Despite their importance, there has been little study of the stratigraphic characteristics of river junctions, or the role of confluence morphodynamics in influencing stratigraphic character and preservation potential. As a result, although it is known that confluences can migrate through time, models of confluence geomorphology and sedimentology are usually presented from the perspective that the confluence remains at a fixed location. This is problematic for a number of reasons, not least of which is the continuing debate over whether it is possible to discriminate between scour that has been generated by autocyclic processes (such as confluence scour) and that driven by allocyclic controls (such as sea-level change). This paper investigates the spatial mobility of river confluences by using the 40-year record of Landsat Imagery to elucidate the styles, rates of change and areal extent over which large river confluence scours may migrate. On the basis of these observations, a new classification of the types of confluence scour is proposed and applied to the Amazon and Ganges-Brahmaputra-Meghna (GBM) basins. This analysis demonstrates that the drivers of confluence mobility are broadly the same as those that drive channel change more generally. Thus in the GBM basin, a high sediment supply, large variability in monsoonal driven discharge and easily erodible bank materials result in a catchment where over 80% of large confluences are mobile over this 40-year window; conversely this figure is < 40% for the Amazon basin. These results highlight that: i) the potential areal extent of confluence scours is much greater than previously assumed, with the location of some confluences on the Jamuna (Brahmaputra) River migrating over a distance of 20 times the tributary channel width; ii) extensive migration in the confluence location is more common than currently assumed, and iii) confluence mobility is often tied to the lithological and hydrological characteristics of the drainage basins that determine sediment yield

The sedimentology of river confluences

Channel confluences are key nodes within large river networks, and yet surprisingly little is known about their spatial and temporal evolution. Moreover, because confluences are associated with vertical scour that typically extends to several times the mean channel depth, the deposits associated with such scours should have a high preservation potential within the rock record. Paradoxically, such scours are rarely observed, and their preservation and sedimentological interpretation are poorly understood. The present study details results from a physically‐based morphodynamic model that is applied to simulate the evolution and alluvial architecture of large river junctions. Boundary conditions within the model were defined to approximate the junction of the Ganges and Jamuna rivers, Bangladesh, with the model output being supplemented by geophysical datasets collected at this junction. The numerical simulations reveal several distinct styles of sedimentary fill that are related to the morphodynamic behaviour of bars, confluence scour downstream of braid bars, bend scour and major junction scour. Comparison with existing, largely qualitative, conceptual models reveals that none of these can be applied simply, although elements of each are evident in the deposits generated by the numerical simulation and observed in the geophysical data. The characteristics of the simulated scour deposits are found to vary according to the degree of reworking caused by channel migration, a factor not considered adequately in current conceptual models of confluence sedimentology. The alluvial architecture of major junction scours is thus characterized by the prevalence of erosion surfaces in conjunction with the thickest depositional sets. Confluence scour downstream of braid bar and bend scour sites may preserve some large individual sets, but these locations are typically characterized by lower average set thickness compared to major junction scour and by a lack of large‐scale erosional surfaces. Areas of deposition not related to any of the specific scour types highlighted above record the thinnest depositional sets. This variety in the alluvial architecture of scours may go some way towards explaining the paradox of ancient junction scours, that while abundant large scours are likely in the rock record, they have been reported rarely. The present results outline the likely range of confluence sedimentology and will serve as a new tool for recognizing and interpreting these deposits in the ancient fluvial record

Gathering at the top? Environmental controls of microplastic uptake and biomagnification in freshwater food webs

Microplastics are ubiquitous in the environment, with high concentrations being detected now also in river corridors and sediments globally. Whilst there has been increasing field evidence of microplastics accumulation in the guts and tissues of freshwater and marine aquatic species, the uptake mechanisms of microplastics into freshwater food webs, and the physical and geological controls on pathway-specific exposures to microplastics, are not well understood. This knowledge gap is hampering the assessment of exposure risks, and potential ecotoxicological and public health impacts from microplastics. This review provides a comprehensive synthesis of key research challenges in analysing the environmental fate and transport of microplastics in freshwater ecosystems, including the identification of hydrological, sedimentological and particle property controls on microplastic accumulation in aquatic ecosystems. This mechanistic analysis outlines the dominant pathways for exposure to microplastics in freshwater ecosystems and identifies potentially critical uptake mechanisms and entry pathways for microplastics and associated contaminants into aquatic food webs as well as their risk to accumulate and biomagnify. We identify seven key research challenges that, if overcome, will permit the advancement beyond current conceptual limitations and provide the mechanistic process understanding required to assess microplastic exposure, uptake, hazard, and overall risk to aquatic systems and humans, and provide key insights into the priority impact pathways in freshwater ecosystems to support environmental management decision making

Neocentromeres Form Efficiently at Multiple Possible Loci in Candida albicans

Centromeres are critically important for chromosome stability and integrity. Most eukaryotes have regional centromeres that include long tracts of repetitive DNA packaged into pericentric heterochromatin. Neocentromeres, new sites of functional kinetochore assembly, can form at ectopic loci because no DNA sequence is strictly required for assembly of a functional kinetochore. In humans, neocentromeres often arise in cells with gross chromosome rearrangements that rescue an acentric chromosome. Here, we studied the properties of centromeres in Candida albicans, the most prevalent fungal pathogen of humans, which has small regional centromeres that lack pericentric heterochromatin. We functionally delimited centromere DNA on Chromosome 5 (CEN5) and then replaced the entire region with the counter-selectable URA3 gene or other marker genes. All of the resulting cen5Δ::URA3 transformants stably retained both copies of Chr5, indicating that a functional neocentromere had assembled efficiently on the homolog lacking CEN5 DNA. Strains selected to maintain only the cen5Δ::URA3 homolog and no wild-type Chr5 homolog also grew well, indicating that neocentromere function is independent of the presence of any wild-type CEN5 DNA. Two classes of neocentromere (neoCEN) strains were distinguishable: “proximal neoCEN” and “distal neoCEN” strains. Neocentromeres in the distal neoCEN strains formed at loci about 200–450 kb from cen5Δ::URA3 on either chromosome arm, as detected by massively parallel sequencing of DNA isolated by CENP-ACse4p chromatin immunoprecipitation (ChIP). In the proximal neoCEN strains, the neocentromeres formed directly adjacent to cen5Δ::URA3 and moved onto the URA3 DNA, resulting in silencing of its expression. Functional neocentromeres form efficiently at several possible loci that share properties of low gene density and flanking repeated DNA sequences. Subsequently, neocentromeres can move locally, which can be detected by silencing of an adjacent URA3 gene, or can relocate to entirely different regions of the chromosome. The ability to select for neocentromere formation and movement in C. albicans permits mechanistic analysis of the assembly and maintenance of a regional centromere

Homeobox Transcription Factors Are Required for Conidiation and Appressorium Development in the Rice Blast Fungus Magnaporthe oryzae

The appropriate development of conidia and appressoria is critical in the disease cycle of many fungal pathogens, including Magnaporthe oryzae. A total of eight genes (MoHOX1 to MoHOX8) encoding putative homeobox transcription factors (TFs) were identified from the M. oryzae genome. Knockout mutants for each MoHOX gene were obtained via homology-dependent gene replacement. Two mutants, ΔMohox3 and ΔMohox5, exhibited no difference to wild-type in growth, conidiation, conidium size, conidial germination, appressorium formation, and pathogenicity. However, the ΔMohox1 showed a dramatic reduction in hyphal growth and increase in melanin pigmentation, compared to those in wild-type. ΔMohox4 and ΔMohox6 showed significantly reduced conidium size and hyphal growth, respectively. ΔMohox8 formed normal appressoria, but failed in pathogenicity, probably due to defects in the development of penetration peg and invasive growth. It is most notable that asexual reproduction was completely abolished in ΔMohox2, in which no conidia formed. ΔMohox2 was still pathogenic through hypha-driven appressoria in a manner similar to that of the wild-type. However, ΔMohox7 was unable to form appressoria either on conidial germ tubes, or at hyphal tips, being non-pathogenic. These factors indicate that M. oryzae is able to cause foliar disease via hyphal appressorium-mediated penetration, and MoHOX7 is mutually required to drive appressorium formation from hyphae and germ tubes. Transcriptional analyses suggest that the functioning of M. oryzae homeobox TFs is mediated through the regulation of gene expression and is affected by cAMP and Ca2+ signaling and/or MAPK pathways. The divergent roles of this gene set may help reveal how the genome and regulatory pathways evolved within the rice blast pathogen and close relatives