Analysis of Microbiome Data.

The microbiome represents a hidden world of tiny organisms populating not only our surroundings but also our own bodies. By enabling comprehensive profiling of these invisible creatures, modern genomic sequencing tools have given us an unprecedented ability to characterize these populations and uncover their outsize impact on our environment and health. Statistical analysis of microbiome data is critical to infer patterns from the observed abundances. The application and development of analytical methods in this area require careful consideration of the unique aspects of microbiome profiles. We begin this review with a brief overview of microbiome data collection and processing and describe the resulting data structure. We then provide an overview of statistical methods for key tasks in microbiome data analysis, including data visualization, comparison of microbial abundance across groups, regression modeling, and network inference. We conclude with a discussion and highlight interesting future directions.

An extended novel approach of river health analysis using nonmetric scaling ordination of fish ecological entities and functional identity indices.

As river ecosystems continue to face environmental pressures, it is crucial to develop and apply new biologically relevant statistical tools to support river health assessments. This study aimed to test the potential significance of nonmetric ordination scores and multidimensional functional indices of fish communities for analyzing river ecological health linked to variations in environmental factors. We conducted a nonmetric multidimensional scaling (NMDS) to derive ordination scores based on twenty fish ecological entities (FEs) that we newly defined, along with their relative abundance (RA). These FEs were determined through a unique combination of trophic, habitat, and tolerance guilds. Additionally, we calculated the functional diversity and identity (F-Ide) indices using the RA-weighted distances between FEs defined by the guild identities in a multidimensional space. The results showed that these novel analyses were helpful in assessing river ecological health and spatial changes in the environment. The dominant F-Ide in each community was largely responsible for the variation of river health scores, driven by the combined impact of chemical water quality, alterations in substrate composition, land use changes, and the gradient of elevation. These environmental factors significantly influenced the site scores in the first axis of NMDS, F-Ide indices, and river health scores. Therefore, our study highlights the practical value of utilizing nonmetric scaling ordination scores of FEs and F-Ide indices as tools for evaluating the health of river ecosystems. By elucidating fish community variance, these tools can aid in determining the extent of river health degradation attributable to environmental stressors, including chemical water pollution and changes in substrate resulting from changes in land use and at different elevations.

Discovering the ecological structure of different macrophyte groups in rivers using non-parametric and parametric multivariate ordination techniques.

This paper analyses various methods of ecological ordering that are often used in modelling the relationship between vegetation and habitat. The results of direct gradient ordination by Canonical correspondence analysis (CCA), which is based on correlation, were compared with Non-metric multidimensional scaling (NMDS), which is based on rank analyses. Both tools were also compared with Detrended correspondence analysis (DCA), which is a popular indirect gradient analysis method. The macrophyte assessment was conducted at 98 river locations in the lowland regions of Poland. Each of the surveyed locations falls within a consistent abiotic category: small to medium-sized lowland rivers with a sandy bottom. Habitat elements analysed included limnological variables and geographic parameters, and the botanical survey focused on submerged macrophytes, including vascular plants, as well as bryophytes and algae. Firstly, it was shown that various analytical tools for determining the importance of ecological factors (Monte Carlo test, BIOENV) identify slightly different significant factors responsible for the development of macrophytes in rivers. Secondly, considerable similarity was found in the structure of macrophyte communities generated on NMDS and DCA biplots, while macrophyte communities were presented very differently based on CCA. Thirdly, the ecological preferences of aquatic plants based on one-dimensional analyses primarily reflected the results of CCA, whereas they did not always follow the ecological pattern revealed by NMDS. Finally, by conducting separate studies for non-vascular plants and vascular macrophytes, it was confirmed that different ecological drivers are responsible for the development of particular groups of macrophytes.

Volatile organic compounds produced during postmortem processes can be linked via chromatographic profiles to individual postmortem bacterial species.

Decomposition odor is produced during postmortem mammalian tissue breakdown by bacteria, insects, and intrinsic chemical processes. Past research has not thoroughly investigated which volatile organic compounds (VOCs) can be linked directly to individual bacterial species on decomposing remains. The purpose of this study was to profile the VOCs produced over time by individual species of bacteria using comprehensive two-dimensional gas chromatography (GC×GC) to expand our foundational knowledge of what each bacterial species contributes to decomposition odor. Five different species of bacteria (Bacillus subtilis, Ignatzschineria indica, Ignatzschineria ureiclastica, Curtobacterium luteum, and Vagococcus lutrae) were cultured on standard nutrient agar individually and monitored daily using solid phase microextraction arrow (SPME Arrow) and GC×GC in combination with quadrupole mass spectrometry (qMS) and flame ionization detection (FID). The GC×GC-qMS/FID approach was used to generate rich VOC profiles that represented the bacterial species' metabolic VOC production longitudinally. The data obtained from the chromatographic output was used to compare with a prior study using one-dimensional GC-qMS, and also between each of the five species to investigate the extent of overlap between species. No single VOC could be found in all five bacterial species investigated, and there was little overlap in the profile between species. To further visualize these differences, chromatographic peak data was investigated using two different ordination strategies, principal component analysis (PCA) and principal coordinate analysis (PCoA). The two ordination strategies were compared with each other using a Procrustes analysis. This was performed to understand differences in ordination strategies between the separation science community and chemical ecological community. Overall, ordination strategies were found to produce similar results, as evidenced by the correlation of PCA and PCoA in the Procrustes analysis. All analysis strategies yielded distinct VOC profiles for each species. Further study of additional species will support understanding of the holistic view of decomposition odor from a chemical ecology perspective, and further support our understanding of the production of decomposition odor that culminates from such a complex environment.

Ecological river health assessments, based on fish ordination analysis of ecological indicator entities and the biological integrity metrics, responding to the chemical water pollution.

Evaluation of the ecological health of rivers requires a focused examination of how biological indicators respond to chemical stressors to offer key insights for effective conservation strategies. We examined the influence of stressors on aquatic ecosystems by analyzing various ecological entities and biotic integrity metrics of fish communities. A nonmetric multidimensional scaling (NMDS) approach was applied to determine scores based on 19 fish ecological entities (FEs) and a fish-based multi-metric index of biotic integrity (mIBI-F). The composition of fish communities in reference clusters differed from the disturbed clusters due to instream chemical stressors. These chemical stressors, including high levels of nutrients, organic matter, and ionic/suspended solids, were linked to variation in the key indicator FEs, whose guild identities were closely associated with instream chemical degradation. The scores of FEs (abundance weighted) and mIBI-F metrics in the first NMDS axis (NMDS1) were significantly linked with chemical health indicators (p < 0.001), such as total phosphorus (R2 = 0.67 and 0.47), electrical conductivity (R2 = 0.59 and 0.49), and chlorophyll-a (R2 = 0.48 and 0.25). These NMDS1 scores showed better accuracy than the conventional mIBI-F score in capturing river ecological health linked with chemical health status as determined by a multi-metric index of water pollution. Our study suggests that based on the ordination approach, the biological integrity of these systems reflected the chemical health.

Oat species and interspecific amphiploids show predominance of diploid nuclei in the syncytial endosperm.

Apart from apomictic types, the Polygonum-type eight-nuclear embryo sac is considered to be dominant in grasses. A triploid endosperm is formed as a result of double fertilisation. This study showed, for the first time, the dominance of diploid nuclei in the syncytial stage of the central cell of embryo sac in oat species and amphiploids. The dominance of diploid nuclei, which were the basis for the formation of polyploid nuclei, was weaker in amphiploids due to aneuploid events. The genomic in situ hybridisation method applied in the study did not distinguish the maternal and paternal haploid nuclei of embryo sac. However, this method demonstrated the lack of a set of genomes of one haploid nucleus. Embryological analyses of the initial stages of oat endosperm development revealed a fertilised egg cell, and two polar nuclei differing in size. It can be assumed that the formation of diploid oat endosperm occurred after the fusion of one polar nucleus and the nucleus of a male gamete, while the second polar nucleus gave rise to 1n nuclei. The levels of ploidy of syncytial nuclei were not influenced by both aneuploid events and correlated with pollen developmental anomalies. The differences in the analysed cytogenetic events distinguished amphiploids and their parental species in the ordination space.

Multivariate study of lice (Insecta: Psocodea: Phthiraptera) assemblages hosted by hummingbirds (Aves: Trochilidae).

Lice were collected from 579 hummingbirds, representing 49 species, in 19 locations in Brazil, Costa Rica, Honduras, Paraguay and Peru, at elevations 0­3000 m above sea level. The following variables were included in an ecological analysis (1) host species' mean body mass, sexual size dimorphism, sexual dichromatism, migratory behaviour and dominance behaviour; (2) mean elevation, mean and predictability of temperature, mean and predictability of precipitation of the host species' geographic area; (3) prevalence and mean abundance of species of lice as measures of infestation. Ordination methods were applied to evaluate data structure. Since the traits are expressed at different scales (nominal, interval and ratio), a principal component analysis based on d-correlations for the traits and a principal coordinates analysis based on the Gower index for species were applied. Lice or louse eggs were found on 80 (13.8%) birds of 22 species. A total of 267 lice of 4 genera, Trochiloecetes, Trochiliphagus, Myrsidea and Leremenopon, were collected, with a total mean intensity of 4.6. There were positive interactions between migration behaviour and infestation indices, with elevational migrants having a higher prevalence and abundance of lice than resident birds. Further, we found weak negative correlations between host body mass and infestation indices and positive correlations between mean elevation and prevalence and abundance of Trochiliphagus. Thus, formerly unknown differences in the ecological characteristics and infestation measures of Trochiliphagus and Trochiloecetes lice were revealed, which allows a better understanding of these associations and their potential impacts on hummingbirds.

Investigating the Mediating Role of Executive Function in the Relationship Between ADHD and DCD Symptoms and Depression in Adults.

This study was designed to test the effects of Attention Deficit Hyperactivity Disorder (ADHD) and Developmental Co-ordination Disorder (DCD) on depression levels and investigate the mediating role of executive function (EF) in adults. Adults with ADHD, DCD, and ADHD + DCD (N = 139) completed self-report measures of ADHD, DCD, depression, and EF. There were distinct profiles of EF across diagnostic groups, and higher depression symptoms in adults with ADHD + DCD than DCD alone. All EF domains were predicted by ADHD symptoms, and several by DCD symptoms. ADHD and DCD symptoms, and most EF domains, predicted depression symptoms. Overall EF difficulties fully mediated the relationships between ADHD/DCD and depression symptoms. Several specific EF domains relating to behavioural regulation and metacognition also showed full/partial mediating effects. The mediating role of EF difficulties between these neurodevelopmental conditions and depression symptoms has implications for their understanding and treatment, suggesting that targeting EF may be important for preventing co-occurring depression.

Policy gaps and food systems optimization: a review of agriculture, environment, and health policies in South Africa.

South Africa faces the triple burden of malnutrition, high poverty levels, unemployment, and inequality. "Wicked problems" such as these require innovative and transdisciplinary responses, multi-stakeholder coordination and collaboration, managing complex synergies and trade-offs, and achieving sustainable outcomes. Through qualitative content analysis of national and provincial sector-based policies, we explored the interlinkages between the agriculture, environment, and health sectors in South Africa in the context of sustainable food and nutrition security and the extent to which these interlinkages are integrated into policy and planning. A systemic analysis of the review outcomes was performed to identify its main learning outcome, the status quo in the policy process. The nature of feedback loops was identified, and a leverage point was suggested. The review highlighted that policymakers in the agriculture, environment and health sectors are aware of, and have understood, the relationships among the three sectors. They have also made attempts to address these interlinkages through collaboration and coordination. Unfortunately, this has been met with several challenges due to fragmented sector-specific mandates and targets and a lack of resources for integrated solutions. This creates implementation gaps and unintended duplication of activities, leading to poor service delivery. Transitioning to sustainable and healthy food systems will only be possible after these gaps have been closed and implementation optimization has been achieved. Focusing on meta-level problem-framing, functional collaboration through transdisciplinary approaches, and integrated targets are critical to successful policy implementation and progressive realization of national goals related to sustainable food and nutrition security, unemployment, poverty, and inequality.

Genome-wide and constrained ordination-based analyses of EC code data support reclassification of the species of Massilia La Scola et al. 2000 into Telluria Bowman et al. 1993, Mokoshia gen. nov. and Zemynaea gen. nov.

Based on genome-wide data, Massilia species belonging to the clade including Telluria mixta LMG 11547T should be entirely transferred to the genus Telluria owing to the nomenclatural priority of the type species Telluria mixta. This results in the transfer of 35 Massilia species to the genus Telluria. The presented data also supports the creation of two new genera since peripherally branching Massilia species are distinct from Telluria and other related genera. It is proposed that 13 Massilia species are transferred to Mokoshia gen. nov. with the type species designated Mokoshia eurypsychrophila comb. nov. The species Massilia arenosa is proposed to belong to the genus Zemynaea gen. nov. as the type species Zemynaea arenosa comb. nov. The genome-wide analysis was well supported by canonical ordination analysis of Enzyme Commission (EC) codes annotated from genomes via pannzer2. This new approach was performed to assess the conclusions of the genome-based data and reduce possible ambiguity in the taxonomic decision making. Cross-validation of EC code data compared within canonical plots validated the reclassifications and correctly visualized the expected genus-level taxonomic relationships. The approach is complementary to genome-wide methodology and could be used for testing sequence alignment based data across genetically related genera. In addition to the proposed broader reclassifications, invalidly described species 'Massilia antibiotica', 'Massilia aromaticivorans', 'Massilia cellulosiltytica' and 'Massilia humi' are described as Telluria antibiotica sp. nov., Telluria aromaticivorans sp. nov., Telluria cellulosilytica sp. nov. and Pseudoduganella humi sp. nov., respectively. In addition, Telluria chitinolytica is reclassified as Pseudoduganella chitinolytica comb. nov. The use of combined genome-wide and annotation descriptors compared using canonical ordination clarifies the taxonomy of Telluria and its sibling genera and provides another way to evaluate complex taxonomic data.

Improved description of terrestrial habitat types by including microbial communities as indicators.

Soils host diverse communities of microorganisms essential for ecosystem functions and soil health. Despite their importance, microorganisms are not covered by legislation protecting biodiversity or habitats, such as the Habitats Directive. Advances in molecular methods have caused breakthroughs in microbial community analysis, and recent studies have shown that parts of the communities are habitat-specific. If distinct microbial communities are present in the habitat types defined in the Habitats Directive, the Directive may be improved by including these communities. Thus, monitoring and reporting of biodiversity and conservation status of habitat types could be based not only on plant communities but also on microbial communities. In the present study, bacterial and plant communities were examined in six habitat types defined in the Habitats Directive by conducting botanical surveys and collecting soil samples for amplicon sequencing across 19 sites in Denmark. Furthermore, selected physico-chemical properties expected to differ between habitat types and explain variations in community composition of bacteria and vegetation were analysed (pH, electrical conductivity (EC), soil texture, soil water repellency, soil organic carbon content (OC), inorganic nitrogen, and in-situ water content (SWC)). Despite some variations within the same habitat type and overlaps between habitat types, habitat-specific communities were observed for both bacterial and plant communities, but no correlation was observed between the alpha diversity of vegetation and bacteria. PERMANOVA analysis was used to evaluate the variables best able to explain variation in the community composition of vegetation and bacteria. Habitat type alone could explain 46% and 47% of the variation in bacterial and plant communities, respectively. Excluding habitat type as a variable, the best model (pH, SWC, OC, fine silt, and Shannon's diversity index for vegetation) could explain 37% of the variation for bacteria. For vegetation, the best model (pH, EC, ammonium content and Shannon's diversity index for bacteria) could explain 25% of the variation. Based on these results, bacterial communities could be included in the Habitats Directive to improve the monitoring, as microorganisms are more sensitive to changes in the environment compared to vegetation, which the current monitoring is based on.

Barriers to Accessing Oncology Services for Effective Cancer Care in the Public Health Institutions in Limpopo Province, South Africa: A Qualitative Study.

It is estimated that by 2030, 24 million people worldwide will develop cancer, and 13 million will die annually, with 75% of deaths in low- and middle-income countries. The management and effective control of care have not been fully achieved due to a lack of material and human resources exacerbated by poor governance and co-ordination of the services. The study aimed to explore barriers to accessing oncology services for effective cancer care in the public health institutions in Limpopo province. The study was conducted in the five district hospitals in Limpopo province. A qualitative exploratory descriptive and contextual approach was used to collect data that employed focus group discussions amongst healthcare professionals in different disciplines. Non-probability purposive sampling was used to sample participants from various sections contributing to oncology care. Five focus group discussions were conducted at the selected hospitals. The data were analysed using the eight steps of Tesch's method. The findings revealed that Limpopo province has a shortage of high-technology medical equipment, poor coordination, and a lack of oncological and allied expertise. Governments should ensure that patients receive the care required as stated in the constitution to navigate cancer care pathways to improve patient health outcomes, particularly in rural areas where care is fragmented and poorly financed. Recommendations to support oncology patients involve psychosocial work and palliative care of the multidisciplinary teams to be put forward. The identified barriers regarding oncology care may contribute to changing the departments' outlook and effective functioning by including interdisciplinary oncology teams at all levels of care.

Temperate forest understory vegetation shifts after 40 years of conservation.

Understanding how vegetation composition and diversity respond to global changes is crucial for effective ecosystem management and conservation. This study evaluated shifts in understory vegetation after 40 years of conservation within Drawa National Park (NW Poland), to check which plant communities changed the most, and whether vegetation shifts reflect global change symptoms (climate change and pollution) or natural forest dynamics. Using ordination and generalized mixed-effects linear models, we assessed changes in alpha diversity metrics, accounting for taxonomic, functional, and phylogenetic aspects within 170 quasi-permanent plots, surveyed in 1973-85 and resurveyed in 2015-19. We found an overall homogenization of forest vegetation and specific shift patterns in certain forest associations. In coniferous and nutrient-poor broadleaved forests, the overall number of species increased due to the replacement of functionally distinct or specialized species with more ubiquitous species that could exploit increased resource availability. In riparian forests and alder carrs we found either shifts from riparian forest to alder carrs or to mesic broadleaved forests. The most stable communities were fertile broadleaved forests. Our study quantified shifts in taxonomic, functional, and phylogenetic diversity after 40 years of conservation and provides important insights into the shifts in vegetation composition in temperate forest communities. In coniferous and nutrient-poor broadleaved forests we found an increase in species richness and replacement of functionally distinct or specialized species by ubiquitous species, indicating increased resource availability. Shifts between wet broadleaved forests and transition into mesic forests suggest water limitation, which can be related to climate change. The most stable were fertile broadleaved forests fluctuating due to natural stand dynamics. The findings highlight the need for ongoing monitoring and management of ecological systems to preserve their diversity and functionality in the face of global changes.

Niche shifts and range expansions after invasions of two major pests: the Asian longhorned beetle and the citrus longhorned beetle.

BACKGROUND: In recent years, the quarantine forestry pests the Asian longhorned beetle (ALB) Anoplophora glabripennis and the citrus longhorned beetle (CLB) Anoplophora chinensis have spread across the Northern Hemisphere, triggering concern about their potential distribution. However, little is known about the niche shifts of the pests during the invasion, making it difficult to assess their potential ranges. We thus employed two distinct approaches (i.e., ordination-based and reciprocal model-based) to compare the native and invaded niches of ALB and CLB after their spread to new continents based on global occurrence records. We further constructed models with pooled occurrences from both the native and invaded ranges to analyze the effects of occurrence partitioning on predicted ranges. RESULTS: We detected expansions in the invaded niches of both pests, indicating that the niches shifted to varying extents after the invasion. Large shares of the native niches of ALB and CLB remained unfilled, revealing the potential for further invasion in new regions. The models calibrated with pooled occurrences clearly underestimated the potential ranges in invaded regions compared with the projections based on partitioned models considering native and invaded areas separately. CONCLUSIONS: These results emphasize the importance of elucidating the niche dynamics of invasive species for obtaining accurately predicted ranges, which may help identify risk areas masked by the assumption of niche conservatism. Furthermore, prevention and quarantine measures for ALB and CLB are clearly needed to avoid future serious damage to forest ecosystems. © 2023 Society of Chemical Industry.

Examining the influence of health sector coordination on the efficiency of county health systems in Kenya.

BACKGROUND: Health systems are complex, consisting of multiple interacting structures and actors whose effective coordination is paramount to enhancing health system goals. Health sector coordination is a potential source of inefficiency in the health sector. We examined how the coordination of the health sector affects health system efficiency in Kenya. METHODS: We conducted a qualitative cross-sectional study, collecting data at the national level and in two purposely selected counties in Kenya. We collected data using in-depth interviews (n = 37) with national and county-level respondents, and document reviews. We analyzed the data using a thematic approach. RESULTS: The study found that while formal coordination structures exist in the Kenyan health system, duplication, fragmentation, and misalignment of health system functions and actor actions compromise the coordination of the health sector. These challenges were observed in both vertical (coordination within the ministry of health, within the county departments of health, and between the national ministry of health and the county department of health) and horizontal coordination mechanisms (coordination between the ministry of health or the county department of health and non-state partners, and coordination among county governments). These coordination challenges are likely to impact the efficiency of the Kenyan health system by increasing the transaction costs of health system functions. Inadequate coordination also impairs the implementation of health programmes and hence compromises health system performance. CONCLUSION: The efficiency of the Kenyan health system could be enhanced by strengthening the coordination of the Kenyan health sector. This can be achieved by aligning and harmonizing the intergovernmental and health sector-specific coordination mechanisms, strengthening the implementation of the Kenya health sector coordination framework at the county level, and enhancing donor coordination through common funding arrangements and integrating vertical disease programs with the rest of the health system. The ministry of health and county departments of health should also review internal organizational structures to enhance functional and role clarity of organizational units and staff, respectively. Finally, counties should consider initiating health sector coordination mechanisms between counties to reduce the fragmentation of health system functions across neighboring counties.

Decision Tree Ensembles Utilizing Multivariate Splits Are Effective at Investigating Beta Diversity in Medically Relevant 16S Amplicon Sequencing Data.

Developing an understanding of how microbial communities vary across conditions is an important analytical step. We used 16S rRNA data isolated from human stool samples to investigate whether learned dissimilarities, such as those produced using unsupervised decision tree ensembles, can be used to improve the analysis of the composition of bacterial communities in patients suffering from Crohn's disease and adenomas/colorectal cancers. We also introduce a workflow capable of learning dissimilarities, projecting them into a lower dimensional space, and identifying features that impact the location of samples in the projections. For example, when used with the centered log ratio transformation, our new workflow (TreeOrdination) could identify differences in the microbial communities of Crohn's disease patients and healthy controls. Further investigation of our models elucidated the global impact amplicon sequence variants (ASVs) had on the locations of samples in the projected space and how each ASV impacted individual samples in this space. Furthermore, this approach can be used to integrate patient data easily into the model and results in models that generalize well to unseen data. Models employing multivariate splits can improve the analysis of complex high-throughput sequencing data sets because they are better able to learn about the underlying structure of the data set. IMPORTANCE There is an ever-increasing level of interest in accurately modeling and understanding the roles that commensal organisms play in human health and disease. We show that learned representations can be used to create informative ordinations. We also demonstrate that the application of modern model introspection algorithms can be used to investigate and quantify the impacts of taxa in these ordinations, and that the taxa identified by these approaches have been associated with immune-mediated inflammatory diseases and colorectal cancer.

The paradox of adaptive trait clines with nonclinal patterns in the underlying genes.

Multivariate climate change presents an urgent need to understand how species adapt to complex environments. Population genetic theory predicts that loci under selection will form monotonic allele frequency clines with their selective environment, which has led to the wide use of genotype-environment associations (GEAs). This study used a set of simulations to elucidate the conditions under which allele frequency clines are more or less likely to evolve as multiple quantitative traits adapt to multivariate environments. Phenotypic clines evolved with nonmonotonic (i.e., nonclinal) patterns in allele frequencies under conditions that promoted unique combinations of mutations to achieve the multivariate optimum in different parts of the landscape. Such conditions resulted from interactions among landscape, demography, pleiotropy, and genetic architecture. GEA methods failed to accurately infer the genetic basis of adaptation under a range of scenarios due to first principles (clinal patterns did not evolve) or statistical issues (clinal patterns evolved but were not detected due to overcorrection for structure). Despite the limitations of GEAs, this study shows that a back-transformation of multivariate ordination can accurately predict individual multivariate traits from genotype and environmental data regardless of whether inference from GEAs was accurate. In addition, frameworks are introduced that can be used by empiricists to quantify the importance of clinal alleles in adaptation. This research highlights that multivariate trait prediction from genotype and environmental data can lead to accurate inference regardless of whether the underlying loci display clinal or nonmonotonic patterns.

Morphological Continua Make Poor Species: Genus-Wide Morphometric Survey of the European Bee Orchids (Ophrys L.).

Despite (or perhaps because of) intensive multidisciplinary research, opinions on the optimal number of species recognised within the Eurasian orchid genus Ophrys range from nine to at least 400. The lower figure of nine macrospecies is based primarily on seeking small but reliable discontinuities in DNA 'barcode' regions, an approach subsequently reinforced and finessed via high-throughput sequencing studies. The upper figure of ca. 400 microspecies reflects the morphological authoritarianism of traditional taxonomy combined with belief in extreme pollinator specificity caused by reliance on pollination through pseudo-copulation, enacted by bees and wasps. Groupings of microspecies that are less inclusive than macrospecies are termed mesospecies. Herein, we present multivariate morphometric analyses based on 51 characters scored for 457 individual plants that together span the full morphological and molecular diversity within the genus Ophrys, encompassing 113 named microspecies that collectively represent all 29 mesospecies and all nine macrospecies. We critique our preferred morphometric approach of accumulating heterogeneous data and analysing them primarily using principal coordinates, noting that our conclusions would have been strengthened by even greater sampling and the inclusion of data describing pseudo-pheromone cocktails. Morphological variation within Ophrys proved to be exceptionally multidimensional, lacking strong directional trends. Multivariate clustering of plants according to prior taxonomy was typically weak, irrespective of whether it was assessed at the level of macrospecies, mesospecies or microspecies; considerable morphological overlap was evident even between subsets of the molecularly differentiable macrospecies. Characters supporting genuine taxonomic distinctions were often sufficiently subtle that they were masked by greater and more positively correlated variation that reflected strong contrasts in flower size, tepal colour or, less often, plant size. Individual macrospecies appear to represent morphological continua, within which taxonomic divisions are likely to prove arbitrary if based exclusively on morphological criteria and adequately sampled across their geographic range. It remains unclear how much of the mosaic of subtle character variation among the microspecies reflects genetic versus epigenetic or non-genetic influences and what proportion of any contrasts observed in gene frequencies can be attributed to the adaptive microevolution that is widely considered to dictate speciation in the genus. Moreover, supplementing weak morphological criteria with extrinsic criteria, typically by imposing constraints on geographic location and/or supposed pollinator preference, assumes rather than demonstrates the presence of even the weakest of species boundaries. Overall, it is clear that entities in Ophrys below the level of macrospecies have insufficiently structured variation, either phenotypic or genotypic, to be resolved into discrete, self-circumscribing ("natural") entities that can legitimately be equated with species as delimited within other less specialised plant genera. Our search for a non-arbitrary (meso)species concept competent to circumscribe an intermediate number of species has so far proven unsuccessful.

Navigating the care of families with a child or children with autistic spectrum disorder.

The aim of this project was to better understand nurse navigators work with children and families who are living with severe autism spectrum disorder to achieve improved health and wellbeing outcomes. Nurse navigators were introduced into the public health sector in Queensland in 2016, with 400 navigators currently working across 16 health services in diverse geographic and demographic settings. Narrative inquiry was used to explore one nurse navigator's journey working with children and families living with severe Autism. The challenges of rigid health systems to adapt to the requirements of children with special needs, particularly in relation to care in the emergency department and where interventional procedures are necessary were apparent. Nurse navigators can effectively co-ordinate the care of an extremely vulnerable patient cohort and provide essential advocacy in a health system that is rigid and lacking the flexibility to deal with individual needs.

Getting the most out of maths: How to coordinate mathematical modelling research to support a pandemic, lessons learnt from three initiatives that were part of the COVID-19 response in the UK.

In March 2020 mathematics became a key part of the scientific advice to the UK government on the pandemic response to COVID-19. Mathematical and statistical modelling provided critical information on the spread of the virus and the potential impact of different interventions. The unprecedented scale of the challenge led the epidemiological modelling community in the UK to be pushed to its limits. At the same time, mathematical modellers across the country were keen to use their knowledge and skills to support the COVID-19 modelling effort. However, this sudden great interest in epidemiological modelling needed to be coordinated to provide much-needed support, and to limit the burden on epidemiological modellers already very stretched for time. In this paper we describe three initiatives set up in the UK in spring 2020 to coordinate the mathematical sciences research community in supporting mathematical modelling of COVID-19. Each initiative had different primary aims and worked to maximise synergies between the various projects. We reflect on the lessons learnt, highlighting the key roles of pre-existing research collaborations and focal centres of coordination in contributing to the success of these initiatives. We conclude with recommendations about important ways in which the scientific research community could be better prepared for future pandemics. This manuscript was submitted as part of a theme issue on "Modelling COVID-19 and Preparedness for Future Pandemics".