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Hormogastrid earthworms are found in the diversity hotspot of the Franco-Iberian domain, together with the better-known family Lumbricidae. Integrative systematics (the combination of morphological, molecular and ecological data) have increased our knowledge of the diversity and evolutionary history of these earthworms, highlighting unresolved taxonomic conflicts. One example of a species group in need of integrative taxonomic revision is the genus Boucheona in France. In this work, we analyzed their diversity using previously published data together with additional data obtained from recently sampled localities. Molecular data including DNA barcodes and additional markers enabled us to reconstruct Bayesian and time-calibrated phylogenies to discuss the evolutionary relationships among the different taxa, and to propose hypotheses regarding their biogeographical history. Based on our results, four species of Boucheona are present in Southern France, including two new taxa. Morphological distinctness and molecular phylogenetics results supported the status of four populations as the newly described Boucheona corbierensis sp. nov., as well as the status of “Hormogaster pretiosa var. nigra” as an independent species, redescribed as Boucheona tenebrae sp. nov. These results provide a new perspective of the importance of the genus Boucheona in southern France, as the possible evolutionary origin of a clade of giant anecic earthworms with unknown (but probably remarkable) impact on ecosystem functioning across their range.

The availability of public genomic resources can greatly assist biodiversity assessment, conservation, and restoration efforts by providing evidence for scientifically informed management decisions. Here we survey the main approaches and applications in biodiversity and conservation genomics, considering practical factors, such as cost, time, prerequisite skills, and current shortcomings of applications. Most approaches perform best in combination with reference genomes from the target species or closely related species. We review case studies to illustrate how reference genomes can facilitate biodiversity research and conservation across the tree of life. We conclude that the time is ripe to view reference genomes as fundamental resources and to integrate their use as a best practice in conservation genomics., Trends in Genetics, 39 (7), ISSN:0168-9525

Since the 18th century, several taxonomists have contributed to knowledge of the French earthworm fauna, one of the best-known earthworm communities in Europe, with 164 species currently described. Surprisingly, new species keep being added constantly, even large ones, which are usually the first to be described. The recent discovery of six undescribed morphospecies of Scherotheca during a sampling survey in southern France suggests that other large Lumbricidae remain to be discovered. We provide here an integrative description of these six species, Scherotheca albina Marchán & Decaëns sp. nov., Scherotheca betharramensis Marchán & Decaëns sp. nov., Scherotheca microthomasi Marchán & Decaëns sp. nov., Scherotheca shaihulud Marchán & Decaëns sp. nov., Scherotheca etcheberryensis Marchán & Decaëns sp. nov. and Scherotheca trezencensis Marchán & Decaëns sp. nov., formally name them and place them in a molecular phylogenetic framework together with their Scherotheca relatives from south-western France. This increases the diversity of Scherotheca to 51 species, including Scherotheca haymozi comb. nov. as a small-sized representative of the genus. The validity of the elevation of several subspecies to specific status was corroborated by molecular phylogenetic evidence, and previously unknown cryptic speciation within Scherotheca was detected. Two geographically coherent clades (roughly matching Nouvelle-Aquitaine and Occitanie) correspond to two existing subgenera. The unusually high taxonomic and morphological diversity of Scherotheca within the Pyrénées-Atlantiques may be related to biotic interactions, climatic factors and historical causes. These features highlight the importance of the conservation of these unique earthworm species.

International audience; The taxonomy of earthworms has been riddled by instability, lack of systematically useful characters, and lax diagnoses of some genera. This has led to the use of some genera such as Allolobophora Eisen, 1874 as taxonomic wastebaskets, blurring their evolution and biogeographical history. The implementation of molecular techniques has revolutionized the systematics of the genus; however, some of its species have not been previously included in molecular phylogenetic analyses. Thus, the molecular markers COI, 16S, ND1, 12S, and 28S were sequenced for six endemic species including several taxa of Allolobophora and Aporrectodea Örely, 1885 (another related catch-all genus). Phylogenetic relationships determined by Bayesian inference and maximum likelihood analyses support the status of two of the six taxa examined (Allolobophora burgondiae Bouché, 1972 and Aporrectodea icterica Savigny, 1826) as part of Allolobophora sensu stricto and a presumed synonymy between Allolobophora and Heraclescolex Qiu and Bouché, 1998. Branch lengths and average pairwise genetic distances support the transfer of Allolobophora satchelli Bouché, 1972 to the genus Panoniona Mršić and Šapkarev, 1988 and the emergence of two new genera, Heraultia gen. nov. and Vosgesia gen. nov., endemic to France, hosting Allolobophora tiginosa Bouché, 1972 and Allolobophora zicsii Bouché, 1972, respectively. The aforementioned changes of status and the diagnosis for Heraultia and Vosgesia are presented. These results provided more evolutionarily and biogeographically coherent earthworm groups and highlighted that the Maghreb and the area around the Alps are potential key locations for the diversification of Allolobophora and several lineages of Lumbricidae.

The invasion of jumping worms, a small group of pheretimoid earthworm species from Asia, has increasingly become an ecological, environmental and conservation issue in forest ecosystems and urban-suburban landscapes around the world. Their presence is often noticed due to their high abundance, distinctive “jumping” behavior, and prominent granular casts on the soil surface. Although they are known to affect soil carbon dynamics and nutrient availability, no single paper has summarized their profound impacts on soil biodiversity, plant community, and animals of all trophic groups that rely on soil and the leaf litter layer for habitat, food, and shelter. In this study, we summarize the biology, invasion, and ecological impacts of invasive jumping worms across North America. We highlight potential impacts of this second wave of earthworm invasion, contrast them with the preceding European earthworm invasion in temperate forests in North America, and identify annual life cycle, reproductive and cocoon survival strategies, casting behavior and co-invasion dynamics as the key factors that contribute to their successful invasion and distinct ecological impacts. We then suggest potential management and control strategies for practitioners and policy makers, underscore the importance of coordinated community science projects in tracking the spread, and identify knowledge gaps that need to be addressed to understand and control the invasion.

Molecular phylogenetics contributes to making important advances in the challenging field of earthworm taxonomy. Use of this type of analysis has enabled clarification of the phylogenetic relationships between early-branching genera of Lumbricidae within the highly diverse Franco-Iberian realm. However, molecular phylogenetic studies of the genus Zophoscolex are scarce and have led to taxonomic uncertainty due to insufficient sampling and the absence of the type species, Z. atlanticus, from such studies. The present study investigated 11 species of Zophoscolex (including Z. atlanticus), and the phylogenetic relationships were deduced from seven molecular markers (COI, COII, 16S, tRNAs, ND1, 12S, 28S) by Bayesian and maximum likelihood inference. The findings show that species of Zophoscolex did not belong to a single clade. Zophoscolex atlanticus was placed in a clade with Z. micellus, Z. graffi and Ethnodrilus zajonci. Other species of the genus were found to belong to the genera Cataladrilus and Compostelandrilus. Finally, most of the Iberian species form a distinct clade, which was formally described as the revised genus Castellodrilus. Based on these findings, Zophoscolex is restricted to French representatives. These results highlight the importance of incorporating type species in molecular phylogenetic analyses in order to reconcile taxonomy and systematics.

Although banned in multiple areas, due to its persistence in the environment, endosulfan constitutes a significant environmental concern. In this work, fourth instar Chironomus riparius larvae were exposed at environmentally relevant endosulfan concentrations of 0.1, 1, and 10 μg/L for 24 h to analyze the possible effects of this acaricide on gene expression and enzymatic activity. Transcriptional changes were studied through the implementation of a real-time polymerase chain reaction array with 42 genes related to several metabolic pathways (endocrine system, detoxification response, stress response, DNA reparation, and immune system). Moreover, glutathione-S-transferase (GST), phenoloxidase (PO), and acetylcholinesterase (AChE) activities were assessed. The five pathways were differentially altered by endosulfan exposure with significant changes in the E93, Dis, MAPR, Met, InR, GSTd3, GSTt3, MRP1, hsp70, hsp40, hsp24, ATM, PARP, Proph, and Def genes. Besides, all of the measured enzymatic activities were modified, with increased activity of GST, followed by PO and AChE. In summary, the results reflected the effects provoked in C. riparius at molecular level despite the absence of lethality. These data raise concerns about the strong alteration on different metabolic routes despite the low concentrations used. Therefore, new risk assessment strategies should consider include the effects at the sub-organismal level as endpoints in addition to the classical ecologically relevant parameters (such as survival). This endeavor will facilitate a comprehensive evaluation of toxicants in the environment.

Progress in genome sequencing now enables the large-scale generation of reference genomes. Various international initiatives aim to generate reference genomes representing global biodiversity. These genomes provide unique insights into genomic diversity and architecture, thereby enabling comprehensive analyses of population and functional genomics, and are expected to revolutionize conservation genomics., Trends in Ecology & Evolution, 37 (3), ISSN:0169-5347, ISSN:1872-8383

A critical question for read across of the hazards of nanomaterials is the degree to which their mechanisms of action differ from those of their bulk chemical constituents. It has been established that metal and metal oxide nanoparticles (NPs) can be accumulated by invertebrate species. Moreover, it has been hypothesised that the observed toxicity resulting on exposure to these nanomaterials is most likely to be associated with the releases of ions from external or internal dissolution leading ultimately to toxicity. However, mechanistic confirmation of the similar modes of action for metal oxide nanomaterials and metal ions in studies invertebrates are largely lacking. Therefore, here we present a toxicogenomic study using exposed individuals of the earthworm Eisenia fetida from a single genetic lineage. We compared gene expression and pathway responses through RNA-seq analysis at equitoxic concentrations (EC50 for reproduction) of ZnO NPs and ionic Zn. We found similar transcriptomic effects for both Zn forms with genetic signatures of tight control of cytosolic Zn concentrations through expression changes of genes encoding several Zn transporters. Activation and regulation of JUN, MAP and JNK kinases indicated a cellular response to the increased Zn concentrations of both forms with compound binding terms also enriched. Our results illustrate the need to consider both toxicokinetic and toxicodynamic mechanisms in the development of adverse outcome pathways for nanomaterials.

Eiseniella neapolitana is a semi-aquatic, diploid earthworm that for many years was related to the cosmopolitan species Eiseniella tetraedra and even considered a subspecies of it. Norealidys andaluciana was described in Spain and is usually synonymized with E. neapolitana. We collected 69 specimens from Italy, Spain, and Cyprus and studied five molecular markers (COI, 16S, 28S, 12S, and ND1) and their morphology to solve this taxonomic problem. Phylogenetic analyses reveal the possible existence of two separate genera confounded under the name Eiseniella, but the study of more molecular markers and species of the genus would be necessary to confirm this. Therefore, the synonymy between Eiseniella and Norealidys is maintained. Various genetic analyses, including species delimitation, confirm the separation between E. neapolitana and E. andaluciana (= N. andaluciana) and excluded that E. neapolitana is a subspecies of E. tetraedra. The resemblance in external appearance despite clear genetic differences of the three species could be explained by convergent adaptation to the aquatic habitat. Despite the expected low haplotype diversity based on the 28S gene, we found a surprisingly high variability in the E. andaluciana (= N. andaluciana) population in Spain. However, its stable predicted secondary structure and its high content of G + C reject the presence of a pseudogene.

Introduction: Group B Streptococcus (GBS) is a gram-positive coccus, that frequently colonizes the human genital and gastrointestinal tract, and is the most common cause of invasive infections in neonates. The high clinical suspicion and early treatment are essential to prevent devastating sequelae and improve the prognosis of these patients. Materials and Methods: A retrospective descriptive study made with children younger than 3 months of age, admitted to a neonatal intensive care unit of a level II hospital, from January 1, 2010 to December 31, 2021, with early (EOI) or late-onset (LOI) neonatal GBS infection. Reviewed obstetric history, risk factors, intrapartum chemoprophylaxis (IPA), clinical manifestations, antibiotic therapy, supportive care, recurrent infection, morbidities and deaths. Results: There were 6 cases registered during 12-year period with GBS infection, 3(50%) had EOI, 3(50%) had LOI and 1 case with recurrent infection (more 2 episodes). Most cases were prematures (4/6; 67%)with average gestational age of 29weeks. Eight GBS infections were documented. In EOI, prematurity and prolonged rupture of membranes without chemoprophylaxis were identified as risk factors. The most common clinical diagnosis was meningitis (62,5%, 5/8) and sepsis/bacteriemia without a focus (37,5%, 3/8). The most used empirical therapy was ampicillin, gentamicin and cefotaxime (50%, 4/8) and the most frequent definitive therapy was Penicillin G. 4 patients needed ventilatory support and 1 needed shock and seizures treatment. In one case hydrocephalus and delayed psychomotor development (DPD) were reported and in another case DPD and eyelid ptosis were also reported. 1 death occurred. Conclusion: In our study, maternal colonization wasn’t a risk factor for EIO, because the 4 mothers who performed GBS culture (2 mothers of term newborns and 2 mothers of preterms, one with invasive disease and other with recurrent infection) were negative. We obtained a higher percentage of meningitis than previously described (62.5% vs 5-10%), probably because most patients were premature and immunosuppressed.

Case Report A short stature 10 year old boy with pes planus, mild hearing loss, failure of exfoliation of the primary dentition, anterior hyperdontia, malocclusion and open anterior fontanelle was observed in Pediatric Orthopedics consultation, and presenting the following alterations (image). Discussion This child has cleidocranial dysplasia (CCD). Is an autosomal dominant disease caused by an heterozygous loss-of-function mutation in the RUNX2 gene, encoding transcription factor CBFA1, on chromosome 6p21, which plays an important role in osteogenesis and differentiation of osteoblasts. The main clinical features include persistently open skull sutures, hypoplasia or aplasia of the clavicles permitting abnormal facility in opposing the shoulders, short middle phalanx of the fifth fingers, dental anomalies (supernumerary teeth, failure of exfoliation of the primary dentition and malocclusion), pes planus, upper respiratory complications, recurrent otitis media, hearing loss and often vertebral malformation (scoliosis). Conclusion The clinical diagnosis of CCD was made during pregnancy, through the detection of aplasia of the clavicles in fetu, in association with the mother having short stature (1.32 meters), persistence of primary teeth and open anterior fontanelle. With 10 years of age, he underwent mendilioma study which revealed the mutation in the RUNX2 gene (c. 747 del (p.(Arg251 Alafs*7))) classified as a pathogenic variant.

Understanding the responses of insect herbivores to plant chemical defences is pivotal for the management of crops and pests. However, the mechanisms of interaction are not entirely understood. In this study, we compared the whole transcriptome gene expression of the aphid Macrosiphum euphorbiae grown on two different varieties of tomato that differ in their inducible chemical defences. We used two isogenic lines of tomato with a shared genetic background that only differ in the presence of type IV glandular trichomes and their associated acylsucrose excretions. This works also reports a de novo transcriptome of the aphid M. euphorbiae. Subsequently, we identified a unique and distinct gene expression profile for the first time corresponding to aphid´s exposure to type IV glandular trichomes and acylsugars. The analysis of the aphid transcriptome shows that tomato glandular trichomes and their associated secretions are highly efficient in triggering stress-related responses in the aphid, and demonstrating that their role in plant defence goes beyond the physical impediment of herbivore activity. Some of the differentially expressed genes were associated with carbohydrate, lipid and xenobiotic metabolisms, immune system, oxidative stress response and hormone biosynthesis pathways. Also, the observed responses are compatible with a starvation syndrome. The transcriptome analysis puts forward a wide range of genes involved in the synthesis and regulation of detoxification enzymes that reveal important underlying mechanisms in the interaction of the aphid with its host plant and provides a valuable genomic resource for future study of biological processes at the molecular level using this aphid.

Small heat shock proteins (sHsps) are oligomeric stress proteins characterized by an α-crystallin domain (ACD). These proteins are localized in different subcellular compartments and play critical roles in the stress physiology of tissues, organs, and whole multicellular eukaryotes. They are ubiquitous proteins found in all living organisms, from bacteria to mammals, but they have never been studied in annelids. Here, a data set of 23 species spanning the annelid tree of life, including mostly transcriptomes but also two genomes, was interrogated and 228 novel putative sHsps were identified and manually curated. The analysis revealed very high protein diversity and showed that a significant number of sHsps have a particular dimeric architecture consisting of two tandemly repeated ACDs. The phylogenetic analysis distinguished three main clusters, two of them containing both monomeric sHsps, and ACDs located downstream in the dimeric sHsps, and the other one comprising the upstream ACDs from those dimeric forms. Our results support an evolutionary history of these proteins based on duplication events prior to the Spiralia split. Monomeric sHsps 76) were further divided into five subclusters. Physicochemical properties, subcellular location predictions, and sequence conservation analyses provided insights into the differentiating elements of these putative functional groups. Strikingly, three of those subclusters included sHsps with features typical of metazoans, while the other two presented characteristics resembling non-metazoan proteins. This study provides a solid background for further research on the diversity, evolution, and function in the family of the sHsps. The characterized annelid sHsps are disclosed as essential for improving our understanding of this important family of proteins and their pleotropic functions. The features and the great diversity of annelid sHsps position them as potential powerful molecular biomarkers of environmental stress for acting as prognostic tool in a diverse range of environments.

Earthworms have a central role in ministering the terrestrial ecosystems and are proving to have an important role in modulating the effects climate change has on soil. Aestivation is a form of dormancy employed by the organisms living in deserts and arid environments, when confronted with prolonged periods of drought. Understanding global metabolic adjustments required for withstanding the harsh conditions of the ever more severe Iberian drought, we performed a global transcriptomic exploration of the endogeic earthworm Carpetania matritensis during aestivation. There were a total of 6352 differentially expressed transcripts in the aestivating group, with 65% being downregulated. Based on GO and KEGG enrichment analyses, downregulated genes seem to be indicative of an overall metabolic depression during aestivation. Indeed we noted a reduction of protein turnover and macromolecule metabolism coupled with suppression of genes involved in digestion. Upregulated genes, namely antioxidant genes and DNA repair genes showed clear signs of abiotic stress caused by ROS generation. Abiotic stress led to transcriptomic changes of genes involved in immune response, mostly affecting the NF-kb signaling pathway as well as changes in apoptotic genes indicating the necessity of investigating these processes in a tissue specific manner. Lastly we uncovered a possible mechanism for water retention by nitrogenous waste accumulation. This study provides the first ever transcriptomic investigation done on aestivating earthworms and as such serves as a general framework for investigation on other earthworm species and other soil invertebrates, which is becoming increasingly important with the current scenario of climate change.

The use of organic Ultraviolet (UV) filters has increased in the last years, either in sunscreens, other cosmetics, or even food packaging. These filters may end up in soil and water since the Wastewater Treatment Plants may not successfully remove them. Among them, benzophenones are known to act as endocrine disruptors. However, most of the studies are directed towards vertebrates and aquatic invertebrates, while there is a lack of information on the molecular mechanisms affected by these compounds on soil dwelling invertebrates. Here, we study the impact of direct acute (48 h) contact of 4-hydroxybenzophenone (4-OHBP) at two sublethal concentrations (0.02 and 0.2 mg/mL) on gene expression of the earthworm Eisenia fetida. Investigated genes were involved in endocrine pathways, stress response, detoxification mechanisms, genotoxicity, energy metabolism and epigenetics. Three of them were identified for the first time in earthworms. Our results suggest that exposure to 4-OHBP affected endocrine pathways, causing an increase in the Ecdysone receptor gene (EcR) expression. Moreover, the UV filter induced changes in the CuZn superoxide dismutase gene (CuZn SOD), indicating an effect in the stress response. Finally, significant changes were detected for glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) expression, indicating that energy metabolism is influenced by the 4-OHBP and highlighting the risks of using GAPDH as an internal reference for Real Time PCR.

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Nowadays, extreme weather events caused by climate change are becoming more frequent. This leads to the occurrence of extreme habitats to which species must adapt. This challenge becomes crucial for species living in unstable environments, such as the riparian earthworm Eiseniella tetraedra. Its cosmopolitan distribution exposes it to various environmental changes, such as freezing in subarctic regions or droughts in Mediterranean areas. Transcriptional changes under cold and desiccation conditions could therefore shed light on the adaptive mechanisms of this species. An experiment was performed for each condition. In the cold experiment, the temperature was lowered to −14 °C ± 2 °C (compared to 8 °C for control samples), and in the desiccation treatment, humidity was lowered from 60% to 15%. Comparisons of gene expression levels between earthworms under freezing conditions and control earthworms revealed a total of 84 differentially expressed genes and comparisons between the desiccation experiment and the control yielded 163 differentially expressed genes. However, no common responses were found between the two treatments. The results suggest that E. tetraedra can acclimate to low temperatures due to the upregulation of genes involved in glucose accumulation. However, downregulation of the respiratory chain suggests that this earthworm does not tolerate freezing conditions. Under desiccation conditions, genes involved in cell protection from apoptosis and DNA repair were upregulated. In contrast, lipid metabolism was downregulated, presumably to conserve resources by reducing the rate at which they are consumed., IS was supported by a Predoctoral Fellowship grant by Universidad Complutense de Madrid, Spain. AV was funded by the European Union's Horizon 2020 research and innovation program through a Marie Sklodowska-Curie individual fellowship (841576). MN was supported by Ramón y Cajal Fellowship (RYC2018-024654-I) and this study was funded by Grant PGC2018-094112-A-I00, both from MCIN/AEI/10.13039/501100011033 and by “ESF: Investing in your future” and “ERDF: A way of making Europe” respectively.

Research on cryptic species complexes has reached a consensus on the necessity of integrating multiple sources of evidence. Reduced representation library techniques such as Genotyping-by-Sequencing (GBS) have proven useful to study these groups. Both integrative taxonomy and genome-wide single nucleotide polymorphism (SNP) data remain to be widely applied to earthworms, an animal group with widespread presence of cryptic diversity. The genus Carpetania (formerly the Hormogaster elisae species complex) was found to contain six deeply divergent genetic lineages and some inconspicuous morphological differentiation based on a handful of Sanger-sequenced markers. Marchán et al. (submitted) delimited three well-supported species-level clades on the basis of a genome-wide SNP dataset and geometric morphometric analyses, highlighting the necessity of a formal taxonomic description of these taxa. In this work, further analyses are applied to the SNP data and a thorough morphological study is performed in order to provide an integrative description of two new species and to redescribe Carpetania elisae. Species-specific SNPs are identified and used as diagnostic characters, and genome-wide and cytochrome oxidase C subunit 1 (COI) genetic distances are compared finding a strong correlation between them. The taxonomic description of these three cryptic species provides a useful tool to include them effectively in ecological studies and biodiversity conservation actions., This work was supported by Universidad Complutense de Madrid and Santander Group under grant Proyecto de Investigación Santander/Complutense PR41/17-21027; Systematics Research Fund (SRF) and Xunta de Galicia. Consellería de Cultura, Educación e Ordenación Universitaria. Secretaria Xeral de Universidades under grant ED431B 2019/038.

AIMS Extra virgin olive oil lowers postprandial glycaemia. We investigated if oleuropein, a component of extra virgin olive oil, exerts a similar effect on postprandial glycaemia and the underlying mechanism. METHODS Twenty healthy subjects were randomly allocated in a cross-over design to 20 mg oleuropein or placebo immediately before lunch. Postprandial glycaemia along with blood insulin, dipeptidyl-peptidase-4 (DPP-4) and glucagon-like peptide-1 and oxidative stress, which included soluble NADPH oxidase-derived peptide activity (sNox2-dp), 8-iso-prostaglandin-2α and platelet p47phox phosphorylation, were analysed before and 2 h after meal. RESULTS After 2 h, subjects who assumed oleuropein had significantly lower blood glucose, DPP-4 activity and higher insulin and glucagon-like peptide-1 compared to placebo. Furthermore, sNox2-dp, 8-iso-PGF2α and platelet p47phox phosphorylation were significantly lower in oleuropein- compared to placebo-treated subjects. DPP-4 significantly correlated with sNox2-dp [Spearman's rho (Rs) = 0.615; P

Xenobiotics such as bisphenol A (BPA), are present in biosolids, which are applied as organic fertilizers in agricultural fields. Their effects on soil life have been poorly assessed, and this is particularly important in the case of earthworms, which represent the main animal biomass in this medium. In the present work we study the impacts of BPA on gene expression of Eisenia fetida, a widely used ecotoxicological model. Chronic soil tests and acute contact tests were performed, and gene expression was analyzed in total tissue and in masculine reproductive organs of the earthworms. The genes studied in this research played a role in endocrine pathways, detoxification mechanisms, stress response, epigenetics, and genotoxicity. Most of the genes were identified for the first time, providing potentially useful biomarkers for future assessments. For chronic exposures, no changes were detected in whole-body tissue; however, masculine reproductive organs showed changes in the expression of genes related to endocrine function (EcR, MAPR, AdipoR), epigenetic mechanisms (DNMTs), genotoxicity (PARP1), and stress responses (HSC70 4). For acute exposures, the expression of one epigenetic-related gene was altered for both whole-body tissues and male reproductive organs (Piwi2). Further changes were detected for whole-body tissues involved in detoxification (Metallothionein), stress (HSC70 4), and genotoxicity (PARP1) mechanisms. Acute exposure effects were also tested in whole-body tissues of juveniles, showing changes in the expression of Metallothionein and Piwi2. The molecular changes found in the analyzed earthworms indicate that exposure to BPA may have negative implications in their populations. Particularly interesting are the alterations related to epigenetic mechanisms, which suggest that future generations may be impacted. This study is the first to evaluate the molecular effects of BPA on soil organisms, and further assays will be necessary to better characterize the true environmental repercussions. Capsule: Levels of gene expression in total-body tissues and masculine reproductive organs were analyzed in earthworms after exposure to bisphenol A and we observed associated changes in detoxification, endocrine, epigenetic, genotoxic and stress pathways.

Oxidative stress plays a pivotal role in inducing endothelial dysfunction and progression from simple fatty liver steatosis (FLD) to non-alcoholic steatohepatitis (NASH). Polyphenols could reduce oxidative stress and restore endothelial function by inhibiting the nicotinamide-adenine-dinucleotide-phosphate (NADPH) oxidase isoform Nox2. The aim of this study was to assess endothelial function and oxidative stress in a population affected by simple FLD and NASH. Furthermore, we analysed the effect of high vs low content of cocoa polyphenols on endothelial function and oxidative stress in patients with NASH.In a cross-sectional study we analysed endothelial function, as assessed by flow-mediated dilation (FMD), and oxidative stress, as assessed by Nox2 activation, serum isoprostanes and nitric oxide bioavailability (NOx), in patients with NASH (n = 19), FLD (n = 19) and controls (n = 19). Then, we performed a randomized, cross-over study in 19 subjects with NASH comparing the effect of 14-days administration of 40 g of chocolate at high (dark chocolate, cocoa85%) versus low content (milk chocolate, cocoa35%) of polyphenols on FMD and oxidative stress. Compared to controls, NASH and FLD patients had higher Nox2 activity and isoprostanes levels and lower FMD and NOx, with a significant gradient between FLD and NASH. The interventional study showed that, compared to baseline, FMD and NOx increased (from 2.9 ± 2.4 to 7.2 ± 3.0% p 0.001 and from 15.9 ± 3.6 to 20.6 ± 4.9 μM, p 0.001, respectively) in subjects given dark but not in those given milk chocolate. A simple linear regression analysis showed that Δ (expressed by difference of values between before and after 14 days of chocolate assumption) of FMD was associated with Δ of Nox2 activity (Rs = -0.323; p = 0.04), serum isoprostanes (Rs: -0.553; p 0.001) and NOx (Rs: 0.557; p 0.001).Cocoa polyphenols improve endothelial function via Nox2 down-regulation in NASH patients.

Two of the main challenges facing soil organisms nowadays are emerging contaminants and climate change. The biosolids used in agriculture can contain xenobiotics, such as bisphenols, not regulated in soil. The proven harmful effect of bisphenol A (BPA) has urged the search for replacements such as bisphenol S (BPS) but no studies have tested the effects of this substitute on soil organisms. Our aim was to study the impacts of BPS on the biological cycle of two earthworm species, Dendrobaena veneta and Eisenia fetida, and compare to earlier results with BPA to confirm its suitability as replacement. Moreover, interactions with temperature were investigated, the increase of which is expected due to climate change. Sublethal concentrations (10, 100, 1000 mg kg−1) were evaluated by an artificial soil test at 21 °C. Additional tests with D. veneta were performed at 26 °C, to observe whether a 5 °C increase affected the toxicity of BPS in earthworms. In E. fetida, reproduction (number of juveniles and cocoons) was negatively affected by BPS but growth was not negatively impacted. In D. veneta reproduction was not affected and greater growth was detected in the lowest concentration tested only at 26 °C. Increase in temperature showed no interaction on BPS toxicity but had a greater negative impact than the toxicant and adults showed significantly lower growth and reproductive rate at 26 °C. A direct contact test with BPS was performed to evaluate mortality and enzymatic activity of glutathione-S-transferase (GST). The measured enzymatic activity was higher with increasing concentration of BPS at 48 h and 21 °C in D. veneta but not in E. fetida or at 26 °C. Our results suggest that BPS is slightly less toxic than BPA supporting its use as alternative and also highlight the harmful effects that an increase in temperature can have in earthworm populations, exceeding those of certain toxicants.

Impaired fasting glucose (IFG) is associated with an increased risk of cardiovascular disease but the underlying mechanisms are still unclear. Aim of the study was to investigate the interplay between platelet activation, lipopolysaccharides (LPS) and markers of oxidative stress in patients with IFG and control subjects.We performed a cross-sectional study including 35 patients with IFG and 35 control subjects who were well comparable for age, sex, body mass index and smoking history. Serum levels of LPS, zonulin (a marker of gut permeability), oxidized LDL and plasma levels of soluble P-selectin, were measured. Patients with IFG had significantly higher levels of sP-selectin, LPS, zonulin and oxLDL compared to control subjects. The IFG status (beta coefficient: 0.518, p 0.001), higher LPS (beta coefficient: 0.352, p = 0.001) and female sex (beta coefficient: 0.179, p = 0.042) were independently associated with higher sP-selectin; in addition, oxLDL was positively associated with sP-selectin (r = 0.530, p 0.001) and LPS (r = 0.529, p = 0.001). In IFG patients, we found a significant association between LPS and zonulin (r = 0.521, p = 0.001); this association was confirmed at multivariable analysis (beta coefficient: 0.512, p = 0.007).Our study provides evidence that patients with IFG have increased platelet activation, and suggests LPS as a potential trigger for in vivo platelet activation in this patient population.

Background: Soluble proprotein convertase subtilisin/kexin type 9 (PCSK9) has been shown to be predictive of cardiovascular events (CVEs) in patients who are at high cardiovascular risk. No...

Little is known about the origin and spatial pattern of the common phenomenon of parthenogenesis in earthworms. Aporrectodea trapezoides is unique in having a global distribution, where asexual forms are broadly known. Here we report two populations in Plasencia (Spain) and Karkra (Algeria) with multiple instances of co-occurrence of sexual and parthenogenetic individuals. We thus studied these populations where the two stages coexist with the aim of understanding the origins of parthenogenesis in earthworms. Two mitochondrial (COII and 16S rRNA) and three nuclear (ITS2, histone H3 and 28S rRNA) genes where evaluated to shed light on the mode, timing and frequency of origin of parthenogenetic forms. A population genetic study showed that in all cases sexual and parthenogenetic earthworms shared mitochondrial and nuclear sequence types, and that there is a strong genetic structure at the geographic level. Genetic divergence in both mitochondrial genes was high (up to 9.61% in COII and 6.12% in 16S rRNA), and ITS2 sequences were identical in individuals from closely related localities within a population. The haplotype networks, AMOVAs, Mantel tests and FST values all support the existence of strong genetic structure at the level of locality, but not due to reproductive mode, therefore providing evidence of multiple recent origins of parthenogenesis in this species.

Mediterranean Diet (Med-Diet) is associated with reduced incidence of vascular events (VEs) in atrial fibrillation (AF), but the mechanism accounting for its beneficial effect is only partially known. We hypothesized that Med-Diet may reduce VEs by improving antioxidant status, as assessed by glutathione peroxidase 3 (GPx3) and superoxide dismutase (SOD). We performed a prospective cohort study investigating the relationship between adherence to Med-Diet, serum baseline GPx3 and SOD activities, and the occurrence of VEs in 690 AF patients. GPx3 activity was directly associated with Med-Diet score (B = 0.192, p 0.001) and inversely with age (B = -0.124, p = 0.001), after adjustment for potential confounders; Med-Diet weakly affected SOD levels. During a mean follow-up of 46.1 ± 28.2 months, 89 VEs were recorded; patients with VEs had lower GPx3 levels compared with those without VEs (p = 0.002); and no differences regarding SOD activity were found. Multivariable Cox regression analysis showed that age (Hazard ratio [HR]:1.065, p 0.001), logGPx3 (above median, HR: 0.629, p 0.05), and Med-Diet score (HR: 0.547, p 0.05) predicted VEs. Med-Diet favorably modulates antioxidant activity of GPx3 in AF, resulting in reduced VEs rate. We hypothesize that the modulation of GPx3 levels by Med-Diet could represent an additional nutritional strategy to prevent VEs in AF patients. Antioxid. Redox Signal. 25, 751-755.

Molecular techniques have accelerated the detection of cryptic diversity within different animal groups, especially in soil fauna. These cryptic taxa show no morphological differences, but other traits vary between them. Research on earthworms has shown different approaches to identify biologically relevant differences between cryptic lineages, which can guide studies on other animals. These can be summarized in ecological preferences, unexplored morphological characters, behavior (and reproductive isolation), further molecular markers, genomic plasticity and epigenetics, and associated biota. These differences highlight the importance of characterizing cryptic genetic lineages in basic and applied research.

Soil organisms, including earthworms, are a key component of terrestrial ecosystems. However, little is known about their diversity, their distribution, and the threats affecting them. We compiled a global dataset of sampled earthworm communities from 6928 sites in 57 countries as a basis for predicting patterns in earthworm diversity, abundance, and biomass. We found that local species richness and abundance typically peaked at higher latitudes, displaying patterns opposite to those observed in aboveground organisms. However, high species dissimilarity across tropical locations may cause diversity across the entirety of the tropics to be higher than elsewhere. Climate variables were found to be more important in shaping earthworm communities than soil properties or habitat cover. These findings suggest that climate change may have serious implications for earthworm communities and for the functions they provide., 677232 to N.E.). K.S.R. and W.H.v.d.P. were supported by ERC-ADV grant 323020 to W.H.v.d.P. Also supported by iDiv (DFG FZT118) Flexpool proposal 34600850 (C.A.G. and N.E.); the Academy of Finland (285882) and the Natural Sciences and Engineering Research Council of Canada (postdoctoral fellowship and RGPIN-2019-05758) (E.K.C.); DOB Ecology (T.W.C., J.v.d.H., and D.R.); ERC-AdG 694368 (M.R.); and the TULIP Laboratory of Excellence (ANR-10-LABX-41) (M.L.). In addition, data collection was funded by the Russian Foundation for Basic Research (12-04-01538-a, 12-04-01734-a, 14-44-03666-r_center_a, 15-29-02724-ofi_m, 16-04-01878-a 19-05-00245); Tarbiat Modares University; Aurora Organic Dairy; UGC(NERO) (F. 1-6/Acctt./NERO/2007-08/1485); Natural Sciences and Engineering Research Council (RGPIN-2017-05391); Slovak Research and Development Agency (APVV-0098-12); Science for Global Development through Wageningen University; Norman Borlaug LEAP Programme and International Atomic Energy Agency (IAEA); São Paulo Research Foundation - FAPESP (12/22510-8); Oklahoma Agricultural Experiment Station; INIA - Spanish Agency (SUM 2006-00012-00-0); Royal Canadian Geographical Society; Environmental Protection Agency (Ireland) (2005-S-LS-8); University of Hawai‘i at Mānoa (HAW01127H; HAW01123M); European Union FP7 (FunDivEurope, 265171); U.S. Department of the Navy, Commander Pacific Fleet (W9126G-13-2-0047); Science and Engineering Research Board (SB/SO/AS-030/2013) Department of Science and Technology, New Delhi, India; Strategic Environmental Research and Development Program (SERDP) of the U.S. Department of Defense (RC-1542); Maranhão State Research Foundation (FAPEMA); Coordination for the Improvement of Higher Education Personnel (CAPES); Ministry of Education, Youth and Sports of the Czech Republic (LTT17033); Colorado Wheat Research Foundation; Zone Atelier Alpes, French National Research Agency (ANR-11-BSV7-020-01, ANR-09-STRA-02-01, ANR 06 BIODIV 009-01); Austrian Science Fund (P16027, T441); Landwirtschaftliche Rentenbank Frankfurt am Main; Welsh Government and the European Agricultural Fund for Rural Development (Project Ref. A AAB 62 03 qA731606); SÉPAQ; Ministry of Agriculture and Forestry of Finland; Science Foundation Ireland (EEB0061); University of Toronto (Faculty of Forestry); National Science and Engineering Research Council of Canada; Haliburton Forest and Wildlife Reserve; NKU College of Arts and Sciences Grant; Österreichische Forschungsförderungsgesellschaft (837393 and 837426); Mountain Agriculture Research Unit of the University of Innsbruck; Higher Education Commission of Pakistan; Kerala Forest Research Institute, Peechi, Kerala; UNEP/GEF/TSBF-CIAT Project on Conservation and Sustainable Management of Belowground Biodiversity; Ministry of Agriculture and Forestry of Finland; Complutense University of Madrid/European Union FP7 project BioBio (FPU UCM 613520); GRDC; AWI; LWRRDC; DRDC; CONICET (National Scientific and Technical Research Council) and FONCyT (National Agency of Scientific and Technological Promotion) (PICT, PAE, PIP), Universidad Nacional de Luján y FONCyT [PICT 2293 (2006)], Fonds de recherche sur la nature et les technologies du Québec (131894), Deutsche Forschungsgemeinschaft [SCHR1000/3-1, SCHR1000/6-1, 6-2 (FOR 1598), WO 670/7-1, WO 670/7-2, and SCHA 1719/1-2], CONACYT (FONDOS MIXTOS TABASCO/PROYECTO11316); NSF (DGE-0549245, DGE-0549245, DEB-BE-0909452, NSF1241932); Institute for Environmental Science and Policy at the University of Illinois at Chicago; Dean’s Scholar Program at UIC; Garden Club of America Zone VI Fellowship in Urban Forestry from the Casey Tree Endowment Fund; J. E. Weaver Competitive Grant from the Nebraska Chapter of The Nature Conservancy; the College of Liberal Arts and Sciences at DePaul University; Elmore Hadley Award for Research in Ecology and Evolution from the UIC Dept. of Biological Sciences; Spanish CICYT (AMB96-1161; REN2000-0783/GLO; REN2003-05553/GLO; REN2003-03989/GLO; CGL2007-60661/BOS); Yokohama National University; MEXT KAKENHI (25220104); Japan Society for the Promotion of Science KAKENHI (25281053, 17KT0074, 25252026); ADEME (0775C0035); Ministry of Science, Innovation and Universities of Spain (CGL2017-86926-P); Syngenta Philippines; UPSTREAM; LTSER (Val Mazia/Matschertal); Marie Sklodowska Curie Postdoctoral Fellowship (747607); National Science and Technology Base Resource Survey Project of China (2018FY100306); McKnight Foundation (14-168); Program of Fundamental Researches of Presidium of Russian Academy of Sciences (AAAA-A18-118021490070-5); Brazilian National Council of Research CNPq; and French Ministry of Foreign and European Affairs. Author contributions: H.R.P.P. led the analysis, data curation, and writing of the original manuscript draft. C.A.G. assisted in analyses and writing of the original manuscript draft. E.K.C. and N.E. revised subsequent manuscript drafts. J.v.d.H., D.R., and T.W.C. provided additional analyses. E.K.C., N.E., and M.P.T. acquired funding for the project. J.K., K.B.G., B.S., M.L.C.B., M.J.I.B., and G.B. contributed to data curation. H.R.P.P., C.A.G., M.L.C.B., M.J.I.B., G.B., O.F., A.O., E.M.B., J.B., U.B., T.D., F.T.d.V., B.K.-R., M.L., J.M., C.M., W.H.v.d.P., K.S.R., M.C.R., D.R., M.R., M.P.T., D.H.W., D.A.W., E.K.C., and N.E. contributed to the project conceptualization. All authors reviewed and edited the final draft manuscript. The majority of the authors provided data for the analyses. Competing interests: The authors declare no competing interests. Data and materials availability: Data and analysis code are available on the iDiv Data repository (DOI: 10.25829/idiv.1804-5-2593) and GitHub (https://github.com/helenphillips/GlobalEWDiversity; DOI: 10.5281/zenodo.3386456).

Stolonization in syllid annelids is a unique mode of reproduction among animals. During the breeding season, a structure resembling the adult but containing only gametes, called stolon, is formed generally at the posterior end of the animal. When stolons mature, they detach from the adult and gametes are released into the water column. The process is synchronized within each species, and it has been reported to be under environmental and endogenous control, probably via endocrine regulation. To further understand reproduction in syllids and to elucidate the molecular toolkit underlying stolonization, we generated Illumina RNA-seq data from different tissues of reproductive and nonreproductive individuals of Syllis magdalena and characterized gene expression during the stolonization process. Several genes involved in gametogenesis (ovochymase, vitellogenin, testis-specific serine/threonine-kinase), immune response (complement receptor 2), neuronal development (tyrosine-protein kinase Src42A), cell proliferation (alpha-1D adrenergic receptor), and steroid metabolism (hydroxysteroid dehydrogenase 2) were found differentially expressed in the different tissues and conditions analyzed. In addition, our findings suggest that several neurohormones, such as methyl farnesoate, dopamine, and serotonin, might trigger stolon formation, the correct maturation of gametes and the detachment of stolons when gametogenesis ends. The process seems to be under circadian control, as indicated by the expression patterns of r-opsins. Overall, our results shed light into the genes that orchestrate the onset of gamete formation and improve our understanding of how some hormones, previously reported to be involved in reproduction and metamorphosis processes in other invertebrates, seem to also regulate reproduction via stolonization. This research received funding from the European Union’s (European Atomic Energy Community’s) Seventh Framework Program (FP7/2007–2013; FP7/2007–2011) under grant agreement 227799 to P.A.-C. Sequencing and analyses were conducted with internal MCZ funds to G.G. and with the support of the Center for Systems Biology and the Research Computing group, both from the Faculty of Arts and Sciences (Harvard University), and with internal funds from the Department of Life Sciences (Natural History Museum of London) to AR. One anonymous reviewer and Associate Editor Mandë Holford provided constructive criticism which helped to improve this study.