Cell imaging by phonon microscopy: sub-optical wavelength ultrasound for non-invasive imaging

The mechanical properties of cells play an important role in cell function and behavior. This paper presents recent developments that have enabled the use of laser-generated phonons (ultrasound) with sub-optical wavelengths to look inside living cells. The phonons reveal contrast from changes in the elasticity of the cell and can provide high resolution three dimensional images

Estimating the extended and hidden species diversity from environmental DNA in hyper-diverse regions

Species inventories are the building blocks of our assessment of biodiversity patterns and human impact. Yet, historical inventories based on visual observations are often incomplete, impairing subsequent analyses of ecological mechanisms, extinction risk and management success. Environmental DNA (eDNA) metabarcoding is an emerging tool that can provide wider biodiversity assessments than classical visual-based surveys. However, eDNA-based inventories remain limited by sampling effort and reference database incompleteness. In this study, we propose a new framework coupling eDNA surveys and sampling-theory methods to estimate species richness in under-sampled and hyper-diverse regions where some species remain absent from the checklist or undetected by visual surveys. We applied this framework to the coastal fish diversity in the heart of the coral triangle, the richest marine biodiversity hotspot worldwide. Combining data from 279 underwater visual censuses, 92 eDNA samples and an extensive custom genetic reference database, we show that eDNA metabarcoding recorded 196 putative species not detected by underwater visual census including 37 species absent from the regional checklist. We provide an updated checklist of marine fishes in the ‘Raja Ampat Bird's Head Peninsula' ecoregion with 2534 species including 1761 confirmed and 773 highly probable presences. The Chao lower-bound diversity estimator, based on the incidence of rare species, shows that the region potentially hosts an additional 123 fish species, including pelagic, cryptobenthic and vulnerable species. The extended and hidden biodiversity along with their asymptotic estimates highlight the ability of eDNA to expand regional inventories and species distributions to better guide conservation strategies

Genetic alterations of ALK in high-risk neuroblastoma patients: a SIOPEN study

Background: In neuroblastoma (NB), the ALK receptor tyrosine kinase can be constitutively activated either through genomic amplification or activating point mutations. We studied ALK genetic alterations in high-risk NB patients to determine their frequency and prognostic impact.N/

Frequency and Prognostic Impact of ALK Amplifications and Mutations in the European Neuroblastoma Study Group (SIOPEN) High-Risk Neuroblastoma Trial (HR-NBL1)

Purpose: In neuroblastoma (NB), the ALK receptor tyrosine kinase can be constitutively activated through activating point mutations or genomic amplification. We studied ALK genetic alterations in high-risk (HR) patients on the HR-NBL1/SIOPEN trial to determine their frequency, correlation with clinical parameters, and prognostic impact. Materials and methods: Diagnostic tumor samples were available from 1,092 HR-NBL1/SIOPEN patients to determine ALK amplification status (n = 330), ALK mutational profile (n = 191), or both (n = 571). Results: Genomic ALK amplification (ALKa) was detected in 4.5% of cases (41 out of 901), all except one with MYCN amplification (MNA). ALKa was associated with a significantly poorer overall survival (OS) (5-year OS: ALKa [n = 41] 28% [95% CI, 15 to 42]; no-ALKa [n = 860] 51% [95% CI, 47 to 54], [P 20% mutated allele fraction) in 10% of cases (76 out of 762) and at a subclonal level (mutated allele fraction 0.1%-20%) in 3.9% of patients (30 out of 762), with a strong correlation between the presence of ALKm and MNA (P < .001). Among 571 cases with known ALKa and ALKm status, a statistically significant difference in OS was observed between cases with ALKa or clonal ALKm versus subclonal ALKm or no ALK alterations (5-year OS: ALKa [n = 19], 26% [95% CI, 10 to 47], clonal ALKm [n = 65] 33% [95% CI, 21 to 44], subclonal ALKm (n = 22) 48% [95% CI, 26 to 67], and no alteration [n = 465], 51% [95% CI, 46 to 55], respectively; P = .001). Importantly, in a multivariate model, involvement of more than one metastatic compartment (hazard ratio [HR], 2.87; P < .001), ALKa (HR, 2.38; P = .004), and clonal ALKm (HR, 1.77; P = .001) were independent predictors of poor outcome. Conclusion: Genetic alterations of ALK (clonal mutations and amplifications) in HR-NB are independent predictors of poorer survival. These data provide a rationale for integration of ALK inhibitors in upfront treatment of HR-NB with ALK alterations.Key Objective: High risk neuroblastoma (HR-NB) is one of the most difficult childhood cancers to cure. This study examined whether the presence of an ALK alteration (amplification or mutation) was associated with a poor prognosis in a large patient series treated on the prospective European high-risk neuroblastoma trial (HR-NBL1). Knowledge Generated: We found that ALK amplification or clonal mutation was associated with inferior prognosis in patients with HR-NB and both are independent prognostic variables on multivariate analysis. To our knowledge, this is the first study to report the highly prognostic significance of ALK amplification in HR-NB. Relevance: As ALK can be targeted therapeutically, this study convincingly argues for the introduction of ALK inhibitors for upfront management of patients with HR-NB with ALK aberrations. Importantly, the prognostic significance of ALK alterations included a subgroup of trial patients treated with the current standard of care for HR-NB including anti-GD2 immunotherapy.info:eu-repo/semantics/publishedVersio

Population dynamics and genetic connectivity in recent chimpanzee history

The European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (grant agreement no. 864203) (to T.M.-B.). BFU2017-86471-P (MINECO/FEDER, UE) (to T.M.-B.). “Unidad de Excelencia María de Maeztu”, funded by the AEI (CEX2018-000792-M) (to T.M.-B.). Howard Hughes International Early Career (to T.M.-B.). NIH 1R01HG010898-01A1 (to T.M.-B.). Secretaria d’Universitats i Recerca and CERCA Program del Departament d’Economia i Coneixement de la Generalitat de Catalunya (GRC 2017 SGR 880) (to T.M.-B.). UCL’s Wellcome Trust ISSF3 award 204841/Z/16/Z (to A.M.A. and J.M.S.). Generalitat de Catalunya (2017 SGR-1040) (to M. Llorente). Wellcome Trust Investigator Award 202802/Z/16/Z (to D.A.H.). The Pan African Program: The Cultured Chimpanzee (PanAf) is generously funded by the Max Planck Society, the Max Planck Society Innovation Fund, and the Heinz L. Krekeler Foundation.Knowledge on the population history of endangered species is critical for conservation, but whole-genome data on chimpanzees (Pan troglodytes) is geographically sparse. Here, we produced the first non-invasive geolocalized catalog of genomic diversity by capturing chromosome 21 from 828 non-invasive samples collected at 48 sampling sites across Africa. The four recognized subspecies show clear genetic differentiation correlating with known barriers, while previously undescribed genetic exchange suggests that these have been permeable on a local scale. We obtained a detailed reconstruction of population stratification and fine-scale patterns of isolation, migration, and connectivity, including a comprehensive picture of admixture with bonobos (Pan paniscus). Unlike humans, chimpanzees did not experience extended episodes of long-distance migrations, which might have limited cultural transmission. Finally, based on local rare variation, we implement a fine-grained geolocalization approach demonstrating improved precision in determining the origin of confiscated chimpanzees.Publisher PDFPeer reviewe

Cross-ocean patterns and processes in fish biodiversity on coral reefs through the lens of eDNA metabarcoding

Increasing speed and magnitude of global change threaten the world's biodiversity and particularly coral reef fishes. A better understanding of large-scale patterns and processes on coral reefs is essential to prevent fish biodiversity decline but it requires new monitoring approaches. Here, we use environmental DNA metabarcoding to reconstruct well-known patterns of fish biodiversity on coral reefs and uncover hidden patterns on these highly diverse and threatened ecosystems. We analysed 226 environmental DNA (eDNA) seawater samples from 100 stations in five tropical regions (Caribbean, Central and Southwest Pacific, Coral Triangle and Western Indian Ocean) and compared those to 2047 underwater visual censuses from the Reef Life Survey in 1224 stations. Environmental DNA reveals a higher (16%) fish biodiversity, with 2650 taxa, and 25% more families than underwater visual surveys. By identifying more pelagic, reef-associated and crypto-benthic species, eDNA offers a fresh view on assembly rules across spatial scales. Nevertheless, the reef life survey identified more species than eDNA in 47 shared families, which can be due to incomplete sequence assignment, possibly combined with incomplete detection in the environment, for some species. Combining eDNA metabarcoding and extensive visual census offers novel insights on the spatial organization of the richest marine ecosystems

The Role of Neutrophil Proteins on the Amyloid Beta-RAGE Axis

We would like to thank Dr. Arthur Owora, previously a Research Biostatistician of the Department of Pharmaceutical Sciences, University of Oklahoma Health Sciences Center, for his assistance on the statistical analysis performed in this study. We thank Dr. Sixia Chen of the Department of Biostatistics and Epidemiogy, University of Oklahoma Health Sciences Center, for his additional input on the statistical analysis. We thank the Laboratory for Molecular Biology and Cytometry Research at the University of Oklahoma Health Sciences Center for the use of the Core Facility which allowed us to perform the MALDI-TOF MS and MS/MS experiments. GM-0111 was provided as a gift by Dr. Justin Savage, GlycoMira Therapeutics, Inc.We previously showed an elevated expression of the neutrophil protein, cationic antimicrobial protein of 37kDa (CAP37), in brains of patients with Alzheimer’s disease (AD), suggesting that CAP37 could be involved in AD pathogenesis. The first step in determining how CAP37 might contribute to AD pathogenesis was to identify the receptor through which it induces cell responses. To identify a putative receptor, we performed GAMMA analysis to determine genes that positively correlated with CAP37 in terms of expression. Positive correlations with ligands for the receptor for advanced glycation end products (RAGE) were observed. Additionally, CAP37 expression positively correlated with two other neutrophil proteins, neutrophil elastase and cathepsin G. Enzyme-linked immunosorbent assays (ELISAs) demonstrated an interaction between CAP37, neutrophil elastase, and cathepsin G with RAGE. Amyloid beta 1–42 (Aβ1–42), a known RAGE ligand, accumulates in AD brains and interacts with RAGE, contributing to Aβ1–42 neurotoxicity. We questioned whether the binding of CAP37, neutrophil elastase and/or cathepsin G to RAGE could interfere with Aβ1–42 binding to RAGE. Using ELISAs, we determined that CAP37 and neutrophil elastase inhibited binding of Aβ1–42 to RAGE, and this effect was reversed by protease inhibitors in the case of neutrophil elastase. Since neutrophil elastase and cathepsin G have enzymatic activity, mass spectrometry was performed to determine the proteolytic activity of all three neutrophil proteins on Aβ1–42. All three neutrophil proteins bound to Aβ1–42 with different affinities and cleaved Aβ1–42 with different kinetics and substrate specificities. We posit that these neutrophil proteins could modulate neurotoxicity in AD by cleaving Aβ1–42 and influencing the Aβ1–42 –RAGE interaction. Further studies will be required to determine the biological significance of these effects and their relevance in neurodegenerative diseases such as AD. Our findings identify a novel area of study that underscores the importance of neutrophils and neutrophil proteins in neuroinflammatory diseases such as AD.Yeshttp://www.plosone.org/static/editorial#pee

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Necessity, potential and limitations of the molecular taxonomic unit approach for analyzing environmental fish DNA biodiversity

La vitesse et l’intensité des changements globaux nécessitent de nouveaux moyens d’observations de la biodiversité qui soient rapides, non-destructifs, standardisés, déployables à large échelle et dans les écosystèmes les plus reculés (océan profond). Les méthodes de recensement classiques reposent sur l’identification morphologique ou sonore des espèces, mais celles-ci sont coûteuses en temps et en expertise. Au-delà de ces signaux, les animaux laissent aussi des traces d’ADN dans leur environnement sous la forme de cellules dermiques, de mucus ou de fèces. Le metabarcoding de cet ADN environnemental (ADNe) consiste à le collecter, l’amplifier et le séquencer pour identifier les espèces présentes grâce à des bases de séquences génétiques de référence. Or, ces bases de référence sont incomplètes, ce qui limite fortement le potentiel de l’ADNe pour révéler la biodiversité présente. Cette thèse a pour but de développer une approche alternative basée sur des unités taxonomiques moléculaires (MOTUs) pour analyser la biodiversité des macroorganismes aquatiques, et plus particulièrement celle des poissons osseux. J’ai tout d’abord réalisé une synthèse globale et spatialisée de la couverture taxonomique des bases de référence de séquences génétiques pour tous les poissons osseux, qui montre une sous-représentation des espèces de la zone tropicale ainsi que des lacunes concernant les espèces menacées et non-indigènes. Seules 13% des espèces de poisson sont séquencées pour le marqueur le plus commun, ce qui exclut toute ambition d’analyse exhaustive de la biodiversité par assignation aux espèces à court et moyen terme. En conséquence, j’ai développé un pipeline bio-informatique pour générer des estimations de la diversité en unités taxonomiques moléculaires (MOTUs) par famille de poissons. Les résultats démontrent que cette diversité en MOTUs représente un excellent proxy de la diversité en espèces à différentes échelles spatiales. Ensuite une application du metabarcoding de l’ADNe et de l’approche en MOTUs a permis d’estimer la diversité fonctionnelle, basée sur les traits des espèces, et la diversité phylogénétique, basée sur l’histoire évolutive des espèces, des poissons tropicaux de manière plus exhaustive que des méthodes traditionnelles (vidéos, plongées). Enfin, dans une première analyse globale de la diversité des récifs coralliens en ADNe, qui rassemble 251 échantillons récoltés depuis l’Océan Indien jusque dans les Caraïbes, l’approche en MOTUs permet de reconstruire les gradients biogéographiques des poissons mais aussi de révéler une hétérogénéité spatiale locale jusqu’alors sous-estimée. Alors qu’il est aujourd’hui crucial de mettre en place des méthodes de suivi efficaces, non dépendantes de spécialistes et à haute fréquence temporelle pour mieux comprendre les effets des changements globaux sur la biodiversité, ces travaux démontrent tout le potentiel de l’ADNe avec approche en MOTUs pour construire des indicateurs robustes de plusieurs facettes de la biodiversité à plusieurs échelles, mais aussi tester les hypothèses théoriques sous-jacentes à la distribution de cette biodiversité.The speed and intensity of global change requires new means of observing biodiversity that are rapid, non-destructive, standardized, widely deployable and in remote ecosystems (deep sea). Conventional inventory methods are based on morphological or acoustic identification of species, which are costly in terms of time and expertise. Beyond these signals, animals also leave traces of DNA in their environment in the form of dermal cells, mucus or feces. The metabarcoding of this environmental DNA (eDNA) consists in collecting this DNA, amplifying and sequencing it to identify the species present using a genetic reference database. However, these reference databases are incomplete, which severely limits the potential of eDNA. The aim of this thesis is to develop an alternative approach based on molecular taxonomic units (MOTUs) to analyze the biodiversity of aquatic macroorganisms, and more particularly that of bony fish. I first performed a global and spatialized synthesis of the taxonomic coverage of the genetic reference database for all bony fishes, which shows an under-representation of species in the tropical zone as well as taxonomic gaps for endangered and non-indigenous species. Only 13% of fish species are sequenced for the most common marker, which excludes any ambition for an exhaustive analysis of biodiversity using only species-level assignments in the short or medium term. Consequently, I have developed a bioinformatics pipeline to generate estimates of diversity using molecular taxonomic units (MOTUs) by fish family. It shows how this MOTU diversity represents an excellent proxy for species diversity at different spatial scales. Then an application of eDNA metabarcoding and the MOTUs approach allowed to estimate the functional diversity, based on species traits, and the phylogenetic diversity, based on the evolutionary history of the species, of tropical fishes in a more exhaustive way than traditional methods (videos, dives). Finally, in a first global analysis of coral reef diversity in eDNA, which brings together 251 samples collected from the Indian Ocean to the Caribbean, the MOTUs approach allows the reconstruction of major trends in fish biogeography but also reveals local spatial heterogeneity hitherto underestimated. While it is now crucial to set up efficient, non-specialist dependent and high temporal frequency monitoring methods to better understand the effects of global changes on biodiversity, this work demonstrates the full potential of eDNA using a MOTUs approach to build robust indicators of several facets of biodiversity at several scales, but also to test theoretical hypotheses underlying the distribution of this biodiversity

Nécessité, potentiel et limitations de l’approche en unités taxonomiques moléculaires pour analyser la biodiversité de l’ADN environnemental des poissons

The speed and intensity of global change requires new means of observing biodiversity that are rapid, non-destructive, standardized, widely deployable and in remote ecosystems (deep sea). Conventional inventory methods are based on morphological or acoustic identification of species, which are costly in terms of time and expertise. Beyond these signals, animals also leave traces of DNA in their environment in the form of dermal cells, mucus or feces. The metabarcoding of this environmental DNA (eDNA) consists in collecting this DNA, amplifying and sequencing it to identify the species present using a genetic reference database. However, these reference databases are incomplete, which severely limits the potential of eDNA. The aim of this thesis is to develop an alternative approach based on molecular taxonomic units (MOTUs) to analyze the biodiversity of aquatic macroorganisms, and more particularly that of bony fish. I first performed a global and spatialized synthesis of the taxonomic coverage of the genetic reference database for all bony fishes, which shows an under-representation of species in the tropical zone as well as taxonomic gaps for endangered and non-indigenous species. Only 13% of fish species are sequenced for the most common marker, which excludes any ambition for an exhaustive analysis of biodiversity using only species-level assignments in the short or medium term. Consequently, I have developed a bioinformatics pipeline to generate estimates of diversity using molecular taxonomic units (MOTUs) by fish family. It shows how this MOTU diversity represents an excellent proxy for species diversity at different spatial scales. Then an application of eDNA metabarcoding and the MOTUs approach allowed to estimate the functional diversity, based on species traits, and the phylogenetic diversity, based on the evolutionary history of the species, of tropical fishes in a more exhaustive way than traditional methods (videos, dives). Finally, in a first global analysis of coral reef diversity in eDNA, which brings together 251 samples collected from the Indian Ocean to the Caribbean, the MOTUs approach allows the reconstruction of major trends in fish biogeography but also reveals local spatial heterogeneity hitherto underestimated. While it is now crucial to set up efficient, non-specialist dependent and high temporal frequency monitoring methods to better understand the effects of global changes on biodiversity, this work demonstrates the full potential of eDNA using a MOTUs approach to build robust indicators of several facets of biodiversity at several scales, but also to test theoretical hypotheses underlying the distribution of this biodiversity.La vitesse et l’intensité des changements globaux nécessitent de nouveaux moyens d’observations de la biodiversité qui soient rapides, non-destructifs, standardisés, déployables à large échelle et dans les écosystèmes les plus reculés (océan profond). Les méthodes de recensement classiques reposent sur l’identification morphologique ou sonore des espèces, mais celles-ci sont coûteuses en temps et en expertise. Au-delà de ces signaux, les animaux laissent aussi des traces d’ADN dans leur environnement sous la forme de cellules dermiques, de mucus ou de fèces. Le metabarcoding de cet ADN environnemental (ADNe) consiste à le collecter, l’amplifier et le séquencer pour identifier les espèces présentes grâce à des bases de séquences génétiques de référence. Or, ces bases de référence sont incomplètes, ce qui limite fortement le potentiel de l’ADNe pour révéler la biodiversité présente. Cette thèse a pour but de développer une approche alternative basée sur des unités taxonomiques moléculaires (MOTUs) pour analyser la biodiversité des macroorganismes aquatiques, et plus particulièrement celle des poissons osseux. J’ai tout d’abord réalisé une synthèse globale et spatialisée de la couverture taxonomique des bases de référence de séquences génétiques pour tous les poissons osseux, qui montre une sous-représentation des espèces de la zone tropicale ainsi que des lacunes concernant les espèces menacées et non-indigènes. Seules 13% des espèces de poisson sont séquencées pour le marqueur le plus commun, ce qui exclut toute ambition d’analyse exhaustive de la biodiversité par assignation aux espèces à court et moyen terme. En conséquence, j’ai développé un pipeline bio-informatique pour générer des estimations de la diversité en unités taxonomiques moléculaires (MOTUs) par famille de poissons. Les résultats démontrent que cette diversité en MOTUs représente un excellent proxy de la diversité en espèces à différentes échelles spatiales. Ensuite une application du metabarcoding de l’ADNe et de l’approche en MOTUs a permis d’estimer la diversité fonctionnelle, basée sur les traits des espèces, et la diversité phylogénétique, basée sur l’histoire évolutive des espèces, des poissons tropicaux de manière plus exhaustive que des méthodes traditionnelles (vidéos, plongées). Enfin, dans une première analyse globale de la diversité des récifs coralliens en ADNe, qui rassemble 251 échantillons récoltés depuis l’Océan Indien jusque dans les Caraïbes, l’approche en MOTUs permet de reconstruire les gradients biogéographiques des poissons mais aussi de révéler une hétérogénéité spatiale locale jusqu’alors sous-estimée. Alors qu’il est aujourd’hui crucial de mettre en place des méthodes de suivi efficaces, non dépendantes de spécialistes et à haute fréquence temporelle pour mieux comprendre les effets des changements globaux sur la biodiversité, ces travaux démontrent tout le potentiel de l’ADNe avec approche en MOTUs pour construire des indicateurs robustes de plusieurs facettes de la biodiversité à plusieurs échelles, mais aussi tester les hypothèses théoriques sous-jacentes à la distribution de cette biodiversité