Species of the common discomycete genus Bisporella reassigned to at least four genera.

Bisporella as typically conceived is a genus of noticeable, bright yellow inoperculate discomycetes. This interpretation of the genus, however, is at odds with Bisporella pallescens, the current name of the type species of the genus; furthermore, the genus has been interpreted as including the unusual species Bisporella resinicola. By comparing morphological and molecular traits of species traditionally included in Bisporella, we show that the genus is polyphyletic, with many "typical" members of the genus belonging instead in Calycina in Pezizellaceae. Bisporella pallescens is conclusively linked with its asexual morph, Bispora antennata, and the genus Bisporella is abandoned as a later synonym of the monotypic genus Bispora (previously applied only to asexual fungi) and placed as sister to Hymenoscyphus in Helotiaceae. Bisporella resinicola is shown to represent an independent monotypic genus, Eustilbum, which so far is placed incertae sedis in Helotiales. Finally, "Bisporella" subpallida, like Bispora, belongs to Helotiaceae but is instead related to "Phaeohelotium" epiphyllum.

Apothecial Ancestry, Evolution, and Re-Evolution in Thelebolales (Leotiomycetes, Fungi).

Closed cleistothecia-like ascomata have repeatedly evolved in non-related perithecioid and apothecioid lineages of lichenized and non-lichenized Ascomycota. The evolution of a closed, darkly pigmented ascoma that protects asci and ascospores is conceived as either an adaptation to harsh environmental conditions or a specialized dispersal strategy. Species with closed ascomata have mostly lost sterile hymenial elements (paraphyses) and the capacity to actively discharge ascospores. The class Leotiomycetes, one of the most speciose classes of Ascomycota, is mainly apothecioid, paraphysate, and possesses active ascospore discharge. Lineages with closed ascomata, and their morphological variants, have evolved independently in several families, such as Erysiphaceae, Myxotrichaceae, Rutstroemiaceae, etc. Thelebolales is a distinctive order in the Leotiomycetes class. It has two widespread families (Thelebolaceae, Pseudeurotiaceae) with mostly closed ascomata, evanescent asci, and thus passively dispersed ascospores. Within the order, closed ascomata dominate and a great diversity of peridia have evolved as adaptations to different dispersal strategies. The type genus, Thelebolus, is an exceptional case of ascomatal evolution within the order. Its species are the most diverse in functional traits, encompassing species with closed ascomata and evanescent asci, and species with open ascomata, active ascospore discharge, and paraphyses. Open ascomata were previously suggested as the ancestral state in the genus, these ascomata depend on mammals and birds as dispersal agents. In this scheme, species with closed ascomata, a lack of paraphyses, and passive ascospore discharge exhibit derived traits that evolved in adaptation to cold ecosystems. Here, we used morphological and phylogenetic methods, as well as the reconstruction of ancestral traits for ascomatal type, asci dehiscence, the presence or absence of paraphyses, and ascospore features to explore evolution within Thelebolales. We demonstrate the apothecial ancestry in Thelebolales and propose a new hypothesis about the evolution of the open ascomata in Thelebolus, involving a process of re-evolution where the active dispersal of ascospores appears independently twice within the order. We propose a new family, Holwayaceae, within Thelebolales, that retains the phenotypic features exhibited by species of Thelebolus, i.e., pigmented capitate paraphyses and active asci discharge with an opening limitation ring.

Which bidomain conductivity is the most important for modelling heart and torso surface potentials during ischaemia?

Mathematical simulations using the bidomain model, which represents cardiac tissue as consisting of an intracellular and an extracellular space, are a key approach that can be used to improve understanding of heart conditions such as ischaemia. However, key inputs to these models, such as the bidomain conductivity values, are not known with any certainty. Since efforts are underway to measure these values, it would be useful to be able to quantify the effect on model outputs of uncertainty in these inputs, and also to determine, if possible, which are the most important values to focus on in experimental studies. Our previous work has systematically studied the sensitivity of heart surface potentials to the bidomain conductivity values, and this was performed using a half-ellipsoidal model of the left ventricle. This study uses a bi-ventricular heart in a torso model and this time looks at the sensitivity of the torso surface potentials, as well as the heart surface potentials, to various conductivity values (blood, torso and the six bidomain conductivities). We found that both epicardial and torso potentials are the most sensitive to the intracellular longitudinal (along the cardiac fibres) conductivity (gil) with more minor sensitivity to the torso conductivity, and that changes in gil have a significant effect on the surface potential distributions on both the torso and the heart.

A modified approach to determine the six cardiac bidomain conductivities.

Accurate values for the six cardiac bidomain conductivities are crucial for meaningful computational studies of conduction in cardiac tissue, and are yet to be determined by experimental means. Although previous studies have proposed an approach using a multi-electrode array to measure potentials, from which the conductivities can be determined, it has been found that the conductivities cannot be retrieved consistently when the noise in the potentials varies. This paper presents a protocol, which not only has been shown to retrieve the conductivities to a reasonable accuracy, but does so under the presence of a more appropriate additive Gaussian noise model, while using fewer computational resources. Through repetitions of the protocol, a comparison of two pre-fabricated 128 electrode arrays, one array with a square arrangement of electrodes and the other with a rectangular arrangement, was made against a 75-electrode array proposed in previous studies. Results indicated that the two pre-fabricated arrays were generally more capable of obtaining the cardiac conductivities to a higher degree of accuracy than the 75-electrode array. The 128-electrode rectangular array was orientated such that the length of the array first ran along the direction of the fibres, then was reorientated such that the length of the array ran perpendicular to the direction of the fibres. The 128-electrode rectangular array, when orientated in this manner, was more capable of retrieving the conductivities than the remainder of the arrays tested, and thus we suggest this arrangement be used during experimental trials.

Brahmaculus gen. nov. (Leotiomycetes, Chlorociboriaceae).

A second genus in Chlorociboriaceae is described here as Brahmaculus gen. nov. Macroscopically distinctive, all species have bright yellow apothecia with several apothecial cups held on short branches at the tip of a long stipe. The genus is widely distributed across the Southern Hemisphere; the four new species described here include two from Chile (B. magellanicus sp. nov., B. osornoensis sp. nov.) and one each from New Zealand (B. moonlighticus sp. nov.) and Australia (B. packhamiae sp. nov.). They differ from species referred to Chlorociboria, the only other genus in Chlorociboriaceae, in their terrestrial habitat and ascomata that are noticeably more hairy than the known Chlorociboria species, most of which have apothecia with short, macroscopically indistinct hair-like elements. Based on our analyses, Chlorociboria as accepted here is paraphyletic. Additional study is needed to clarify where alternative, monophyletic generic limits should be drawn and how these genera may be recognised morphologically. Also described here are three new Chlorociboria spp. from New Zealand (C. metrosideri sp. nov., C. solandri sp. nov., C. subtilis sp. nov.), distinctive in developing on dead leaves rather than wood and in two of them not forming the green pigmentation characteristic of most Chlorociboria species. New Zealand specimens previously incorrectly identified as Chlorociboria argentinensis are provided with a new name, C. novae-zelandiae sp. nov.

Fungal taxonomy and sequence-based nomenclature.

The identification and proper naming of microfungi, in particular plant, animal and human pathogens, remains challenging. Molecular identification is becoming the default approach for many fungal groups, and environmental metabarcoding is contributing an increasing amount of sequence data documenting fungal diversity on a global scale. This includes lineages represented only by sequence data. At present, these taxa cannot be formally described under the current nomenclature rules. By considering approaches used in bacterial taxonomy, we propose solutions for the nomenclature of taxa known only from sequences to facilitate consistent reporting and communication in the literature and public sequence repositories.

Approaches for determining cardiac bidomain conductivity values: progress and challenges.

Modelling the electrical activity of the heart is an important tool for understanding electrical function in various diseases and conduction disorders. Clearly, for model results to be useful, it is necessary to have accurate inputs for the models, in particular the commonly used bidomain model. However, there are only three sets of four experimentally determined conductivity values for cardiac ventricular tissue and these are inconsistent, were measured around 40 years ago, often produce different results in simulations and do not fully represent the three-dimensional anisotropic nature of cardiac tissue. Despite efforts in the intervening years, difficulties associated with making the measurements and also determining the conductivities from the experimental data have not yet been overcome. In this review, we summarise what is known about the conductivity values, as well as progress to date in meeting the challenges associated with both the mathematical modelling and the experimental techniques. Graphical abstract Epicardial potential distributions, arising from a subendocardial ischaemic region, modelled using conductivity data from the indicated studies.

Unambiguous identification of fungi: where do we stand and how accurate and precise is fungal DNA barcoding?

True fungi (Fungi) and fungus-like organisms (e.g. Mycetozoa, Oomycota) constitute the second largest group of organisms based on global richness estimates, with around 3 million predicted species. Compared to plants and animals, fungi have simple body plans with often morphologically and ecologically obscure structures. This poses challenges for accurate and precise identifications. Here we provide a conceptual framework for the identification of fungi, encouraging the approach of integrative (polyphasic) taxonomy for species delimitation, i.e. the combination of genealogy (phylogeny), phenotype (including autecology), and reproductive biology (when feasible). This allows objective evaluation of diagnostic characters, either phenotypic or molecular or both. Verification of identifications is crucial but often neglected. Because of clade-specific evolutionary histories, there is currently no single tool for the identification of fungi, although DNA barcoding using the internal transcribed spacer (ITS) remains a first diagnosis, particularly in metabarcoding studies. Secondary DNA barcodes are increasingly implemented for groups where ITS does not provide sufficient precision. Issues of pairwise sequence similarity-based identifications and OTU clustering are discussed, and multiple sequence alignment-based phylogenetic approaches with subsequent verification are recommended as more accurate alternatives. In metabarcoding approaches, the trade-off between speed and accuracy and precision of molecular identifications must be carefully considered. Intragenomic variation of the ITS and other barcoding markers should be properly documented, as phylotype diversity is not necessarily a proxy of species richness. Important strategies to improve molecular identification of fungi are: (1) broadly document intraspecific and intragenomic variation of barcoding markers; (2) substantially expand sequence repositories, focusing on undersampled clades and missing taxa; (3) improve curation of sequence labels in primary repositories and substantially increase the number of sequences based on verified material; (4) link sequence data to digital information of voucher specimens including imagery. In parallel, technological improvements to genome sequencing offer promising alternatives to DNA barcoding in the future. Despite the prevalence of DNA-based fungal taxonomy, phenotype-based approaches remain an important strategy to catalog the global diversity of fungi and establish initial species hypotheses.

Three new species and a new combination of Triblidium.

Triblidiaceae (Rhytismatales) currently consists of two genera: Triblidium and Huangshania. Triblidium is the type genus and is characterised by melanized apothecia that occur scattered or in small clusters on the substratum, cleistohymenial (opening in the mesohymenial phase), inamyloid thin-walled asci and hyaline muriform ascospores. Before this study, only the type species, Triblidium caliciiforme, had DNA sequences in the NCBI GenBank. In this study, six specimens of Triblidium were collected from China and France and new ITS, mtSSU, LSU and RPB2 sequences were generated. Our molecular phylogenetic analysis and morphological study demonstrated three new species of Triblidium, which are formally described here: T. hubeiense, T. rostriforme and T. yunnanense. Additionally, our results indicated that Huangshania that was considered to be distinct from Triblidium because of its elongated, transversely-septate ascospores, is congeneric with Triblidium. Therefore, we have placed Huangshania in synonymy under Triblidium, rendering Triblidiaceae a monotypic family.

Differences between models of partial thickness and subendocardial ischaemia in terms of sensitivity analyses of ST-segment epicardial potential distributions.

Mathematical modelling is a useful technique to help elucidate the connection between non-transmural ischaemia and ST elevation and depression of the ECG. Generally, models represent non-transmural ischaemia using an ischaemic zone that extends from the endocardium partway to the epicardium. However, recent experimental work has suggested that ischaemia typically arises within the heart wall. This work examines the effect of modelling cardiac ischaemia in the left ventricle using two different models: subendocardial ischaemia and partial thickness ischaemia, representing the first and second scenarios, respectively. We found that it is possible, only in the model of subendocardial ischaemia, to see a single minimum on the epicardial surface above the ischaemic region, and this only occurs for low ischaemic thicknesses. This may help to explain the rarity of ST depression that is located over the ischaemic region. It was also found that, in both models, the epicardial potential distribution is most sensitive to the proximity of the ischaemic region to the epicardium, rather than to the thickness of the ischaemic region. Since proximity does not indicate the thickness of the ischaemic region, this suggests a reason why it may be difficult to determine the degree of ischaemia using the ST segment of the ECG.

A multigene phylogeny toward a new phylogenetic classification of Leotiomycetes.

Fungi in the class Leotiomycetes are ecologically diverse, including mycorrhizas, endophytes of roots and leaves, plant pathogens, aquatic and aero-aquatic hyphomycetes, mammalian pathogens, and saprobes. These fungi are commonly detected in cultures from diseased tissue and from environmental DNA extracts. The identification of specimens from such character-poor samples increasingly relies on DNA sequencing. However, the current classification of Leotiomycetes is still largely based on morphologically defined taxa, especially at higher taxonomic levels. Consequently, the formal Leotiomycetes classification is frequently poorly congruent with the relationships suggested by DNA sequencing studies. Previous class-wide phylogenies of Leotiomycetes have been based on ribosomal DNA markers, with most of the published multi-gene studies being focussed on particular genera or families. In this paper we collate data available from specimens representing both sexual and asexual morphs from across the genetic breadth of the class, with a focus on generic type species, to present a phylogeny based on up to 15 concatenated genes across 279 specimens. Included in the dataset are genes that were extracted from 72 of the genomes available for the class, including 10 new genomes released with this study. To test the statistical support for the deepest branches in the phylogeny, an additional phylogeny based on 3156 genes from 51 selected genomes is also presented. To fill some of the taxonomic gaps in the 15-gene phylogeny, we further present an ITS gene tree, particularly targeting ex-type specimens of generic type species. A small number of novel taxa are proposed: Marthamycetales ord. nov., and Drepanopezizaceae and Mniaeciaceae fams. nov. The formal taxonomic changes are limited in part because of the ad hoc nature of taxon and specimen selection, based purely on the availability of data. The phylogeny constitutes a framework for enabling future taxonomically targeted studies using deliberate specimen selection. Such studies will ideally include designation of epitypes for the type species of those genera for which DNA is not able to be extracted from the original type specimen, and consideration of morphological characters whenever genetically defined clades are recognized as formal taxa within a classification.

Accuracy of electrocardiographic imaging using the method of fundamental solutions.

Solving the inverse problem of electrocardiology via the Method of Fundamental Solutions has been proposed previously. The advantage of this approach is that it is a meshless method, so it is far easier to implement numerically than many other approaches. However, determining the heart surface potential distribution is still an ill-posed problem and thus requires some form of Tikhonov regularisation to obtain the required distributions. In this study, several methods for determining an "optimal" regularisation parameter are compared in the context of solving the inverse problem of electrocardiology via the Method of Fundamental Solutions. It is found that the Robust Generalised Cross-Validation method most often yields epicardial potential distributions with the least relative error when compared to the input distribution. The study also compares the inverse solutions obtained with the Method of Fundamental Solutions with those obtained in a previous study using the boundary element method. It is found that choosing the best solution methodology and regularisation parameter determination method depends on the particular scenario being considered.

Sensitivity analysis of ST-segment epicardial potentials arising from changes in ischaemic region conductivities in early and late stage ischaemia.

Although computational studies are increasingly used to gain insight into diseases such as myocardial ischaemia, there is still considerable uncertainty about the values for many of the parameters in these studies. This is particularly true for the bidomain conductivity values that are used in normal tissue and, even more so, in ischaemic tissue, when modelling ischaemia. In this work, we extended a previous study that used a half-ellipsoidal model and a realistic model to study subendocardial ischaemia during the ST segment, so that we could simulate both early and late stage ischaemia. We found that, for both stages of ischaemia, there was still the same connection between the degree of ischaemia and the development of features such as minima and maxima in the epicardial potential distribution (EPD), although the magnitudes of the potentials were very often less, which may be significant in terms of detecting them experimentally. Using uncertainty quantification associated with the ischaemic region conductivities, we also determined that the EPD features were sensitive to the ischaemic region extracellular normal and longitudinal conductivities during early stage ischaemia, whereas, during late stage ischaemia, the intracellular longitudinal conductivity was the most significant. However, since we again found that these effects were minor compared with the effects of fibre rotation angle and ischaemic depth, this might suggest that it is not necessary to use different conductivity values inside and outside the ischaemic region when modelling ST segment subendocardial ischaemia, unless the magnitudes of the potentials are an important part of the study.

Import volumes and biosecurity interventions shape the arrival rate of fungal pathogens.

Global trade and the movement of people accelerate biological invasions by spreading species worldwide. Biosecurity measures seek to allow trade and passenger movements while preventing incursions that could lead to the establishment of unwanted pests, pathogens, and weeds. However, few data exist to evaluate whether changes in trade volumes, passenger arrivals, and biosecurity measures have altered rates of establishment of nonnative species over time. This is particularly true for pathogens, which pose significant risks to animal and plant health and are consequently a major focus of biosecurity efforts but are difficult to detect. Here, we use a database of all known plant pathogen associations recorded in New Zealand to estimate the rate at which new fungal pathogens arrived and established on 131 economically important plant species over the last 133 years. We show that the annual arrival rate of new fungal pathogens increased from 1880 to about 1980 in parallel with increasing import trade volume but subsequently stabilised despite continued rapid growth in import trade and recent rapid increases in international passenger arrivals. Nevertheless, while pathogen arrival rates for crop and pasture species have declined in recent decades, arrival rates have increased for forestry and fruit tree species. These contrasting trends between production sectors reflect differences in biosecurity effort and suggest that targeted biosecurity can slow pathogen arrival and establishment despite increasing trade and international movement of people.

Determining the most significant input parameters in models of subendocardial ischaemia and their effect on ST segment epicardial potential distributions.

There is considerable interest in simulating ischaemia in the ventricle and its effect on the electrocardiogram, because a better understanding of the connection between the two may lead to improvements in diagnosis of myocardial ischaemia. In this work we studied subendocardial ischaemia, in a simplified half-ellipsoidal bidomain model of a ventricle, and its effect on ST segment epicardial potential distributions (EPDs). We found that the EPD changed as the ischaemic depth increased, from a single minimum (min1) over the ischaemic region to a maximum (max) there, with min1 over the border of the region. Lastly, a second minimum (min2) developed on the opposite side of the ischaemic region, in addition to min1 and max. We replicated these results in a realistic ventricular model and showed that the min1 only case could be found for ischaemic depths of up to around 35% of the ventricular wall. In addition, we systematically examined the sensitivity of EPD parameters, such as the potentials and positions of min1, max and min2, to various inputs to the half-ellipsoidal model, such as fibre rotation angle, ischaemic depth and conductivities. We found that the EPD parameters were not sensitive to the blood or transverse bidomain conductivities and were most sensitive to either ischaemic depth and/or fibre rotation angle. This allowed us to conclude that the asynchronous development of the two minima might provide a way of distinguishing between low and high thickness subendocardial ischaemia, and that this method may well be valid despite variability in the population.

Quantifying the effect of uncertainty in input parameters in a simplified bidomain model of partial thickness ischaemia.

Reduced blood flow in the coronary arteries can lead to damaged heart tissue (myocardial ischaemia). Although one method for detecting myocardial ischaemia involves changes in the ST segment of the electrocardiogram, the relationship between these changes and subendocardial ischaemia is not fully understood. In this study, we modelled ST-segment epicardial potentials in a slab model of cardiac ventricular tissue, with a central ischaemic region, using the bidomain model, which considers conduction longitudinal, transverse and normal to the cardiac fibres. We systematically quantified the effect of uncertainty on the input parameters, fibre rotation angle, ischaemic depth, blood conductivity and six bidomain conductivities, on outputs that characterise the epicardial potential distribution. We found that three typical types of epicardial potential distributions (one minimum over the central ischaemic region, a tripole of minima, and two minima flanking a central maximum) could all occur for a wide range of ischaemic depths. In addition, the positions of the minima were affected by both the fibre rotation angle and the ischaemic depth, but not by changes in the conductivity values. We also showed that the magnitude of ST depression is affected only by changes in the longitudinal and normal conductivities, but not by the transverse conductivities.

Overview of Phacidiales, including Aotearoamyces gen. nov. on Nothofagus.

The new genus Aotearoamyces is proposed to accommodate a single species that was repeatedly collected on fallen wood in Nothofagaceae forests of New Zealand and was previously misidentified as a Claussenomyces species. This monotypic genus belongs to Tympanidaceae, a recently erected family in Phacidiales. Aotearoamyces is differentiated from other Tympanidaceae by phragmospores that do not form conidia either in or outside the asci, an exciple of textura intricata with hyphae widely spaced and strongly gelatinized (plectenchyma), and apically flexuous, partly helicoid paraphyses. The asexual morph was studied in pure culture. Phylogenetic analyses of combined SSU, ITS and LSU sequences strongly support a sister relationship between the sexually typified Aotearoamyces and the asexually typified "Collophorina" paarla characterized morphologically by forming endoconidia, a feature not found in the genetically distinct type species of Collophorina. Based on our molecular results, we place the genus Epithamnolia in the Mniaecia lineage within Phacidiales.

Comparing diversity of fungi from living leaves using culturing and high-throughput environmental sequencing.

High-throughput sequencing technologies using amplicon approaches have changed the way that studies investigating fungal distribution are undertaken. These powerful and time-efficient technologies have the potential for the first time to accurately map fungal distributions across landscapes or changes in diversity across ecological or biological gradients of interest. There is no requirement for a fungus to form a fruiting body to be detected, and both culturable and nonculturable organisms can be detected. Here we use high-throughput amplicon sequencing from bulk DNA extracts to test the impact that biases associated with culture-based methods had on an earlier study that compared the influence of site and host on fungal diversity in Nothofagaceae forests in New Zealand. Both detection methods sampled tissue from the same set of symptomless, living leaves. We found that both the culturing and high-throughput approaches show that host is a stronger driver of fungal community structure than site, but that both methods have some taxonomic biases. We also found that the individual trees selected for high-throughput sampling can impact the alpha-diversity detected and through this could potentially affect subsequent analyses based on a comparison of this diversity.

The effect of boundary conditions on epicardial potential distributions.

This study presents a comparison of semi-analytical and numerical solution techniques for solving the passive bidomain equation in simple tissue geometries containing a region of subendocardial ischaemia. When the semi-analytical solution is based on Fourier transforms, recovering the solution from the frequency domain via fast Fourier transforms imposes a periodic boundary condition on the solution of the partial differential equation. On the other hand, the numerical solution uses an insulation boundary condition. When these techniques are applied to calculate the epicardial surface potentials, both yield a three well potential distribution which is identical if fibre rotation within the tissue is ignored. However, when fibre rotation is included, the resulting three-well distribution rotates, but through different angles, depending on the solution method. A quantitative comparison between the semi-analytical and numerical solutiontechniques is presented in terms of the effect fibre rotation has on the rotation of the epicardial potential distribution. It turns out that the Fourier transform approach predicts a larger rotation of the epicardial potential distribution than the numerical solution. The conclusion from this study is that it is not always possible to use analytical or semi-analytical solutions to check the accuracy of numerical solution procedures. For the problem considered here, this checking is only possible when it is assumed that there is no fibre rotation through the tissue.