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This paper reports an immunofluorescence confocal microscopy study of the presence and distribution of muscarinic acetylcholine receptor subtypes M1–M5 family in somatic muscle cells and cholinergic synapses of the body wall of the earthworm Lumbricus terrestris. It has been demonstrated that all subtypes of receptors have uneven distribution patterns in the extrasynaptic zones of muscle cells and are also present in the end plate zones. Receptors belonging to subtypes М2–М5, but not to the M1 subtype, can be detected also in the presynaptic membranes of motor nerve terminals. [ABSTRACT FROM AUTHOR]

Background: Methylation of cytosines in DNA (5mC methylation) is a major epigenetic modification that modulates gene expression and constitutes the basis for mechanisms regulating multiple aspects of embryonic development and cell reprogramming in vertebrates. In mammals, 5mC methylation of promoter regions is linked to transcriptional repression. Transcription regulation by 5mC methylation notably involves the nucleosome remodeling and deacetylase complex (NuRD complex) which bridges DNA methylation and histone modifications. However, less is known about regulatory mechanisms involving 5mC methylation and their function in non-vertebrate animals. In this paper, we study 5mC methylation in the marine annelid worm Platynereis dumerilii, an emerging evolutionary and developmental biology model capable of regenerating the posterior part of its body post-amputation. Results: Using in silico and experimental approaches, we show that P. dumerilii displays a high level of DNA methylation comparable to that of mammalian somatic cells. 5mC methylation in P. dumerilii is dynamic along the life cycle of the animal and markedly decreases at the transition between larval to post-larval stages. We identify a full repertoire of mainly single-copy genes encoding the machinery associated with 5mC methylation or members of the NuRD complex in P. dumerilii and show that this repertoire is close to the one inferred for the last common ancestor of bilaterians. These genes are dynamically expressed during P. dumerilii development and regeneration. Treatment with the DNA hypomethylating agent Decitabine impairs P. dumerilii larval development and regeneration and has long-term effects on post-regenerative growth. Conclusions: Our data reveal high levels of 5mC methylation in the annelid P. dumerilii, highlighting that this feature is not specific to vertebrates in the bilaterian clade. Analysis of DNA methylation levels and machinery gene expression during development and regeneration, as well as the use of a chemical inhibitor of DNA methylation, suggest an involvement of 5mC methylation in P. dumerilii development and regeneration. We also present data indicating that P. dumerilii constitutes a promising model to study biological roles and mechanisms of DNA methylation in non-vertebrate bilaterians and to provide new knowledge about evolution of the functions of this key epigenetic modification in bilaterian animals. [ABSTRACT FROM AUTHOR]

We present an updated checklist of marine annelids from the Indian subcontinent. Records of annelid species were obtained from published and grey literatures and online databases. Our review of annelid publications was restricted to the Indian continental shelf region. This paper also discusses the taxonomic status of marine annelid species recorded from this region and problems that impede its research. The updated list comprises of 727 species belonging to 334 genera and 72 families. A total of 152 species have their type locality in India including 88 species that are endemic to the region. The current checklist indicates that 25% (183 species) of the records are questionable and require further examination. Our results highlight that marine annelid richness of the Indian sub-continent is underestimated with many of the native undescribed species being most likely concealed under 'erroneous' or 'cosmopolitan' records. With a combination of factors that include a lack of experts, funding, and failure to update regional literature has resulted in an incomplete state of knowledge for the marine annelid biodiversity from this region. Therefore, there is an urgent need for extensive and intensive sampling to discover new species, conduct detailed re-examinations of doubtful records and, collaborate within the local and international institutes and organizations to improve the regional biodiversity studies. [ABSTRACT FROM AUTHOR]

Earthworms are primary candidates for national soil health monitoring as they are ecosystem engineers that benefit both food production and ecosystem services associated with soil security. Supporting farmers to monitor soil health could help to achieve the policy aspiration of sustainable soils by 2030 in England; however, little is known about how to overcome participation barriers, appropriate methodologies (practical, cost-effective, usefulness) or training needs. This paper presents the results from a pilot #60minworms study which mobilised farmers to assess over >1300 ha farmland soils in spring 2018. The results interpretation framework is based on the presence of earthworms from each of the three ecological groups at each observation (20 x 20 cm x 20 cm pit) and spatially across a field (10 soil pits). Results showed that most fields have basic earthworm presence and abundance, but 42% fields may be over-worked as indicated by absence/rarity of epigeic and/or anecic earthworms. Tillage had a negative impact (p < 0.05) on earthworm populations and organic matter management did not mitigate tillage impacts. In terms of farmer participation, Twitter and Farmers Weekly magazine were highly effective channels for recruitment. Direct feedback from participants included excellent scores in trust, value and satisfaction of the protocol (e.g. 100% would do the test again) and 57% would use their worm survey results to change their soil management practices. A key training need in terms of earthworm identification skills was reported. The trade-off between data quality, participation rates and fieldwork costs suggests there is potential to streamline the protocol further to #30minworms (5 pits), incurring farmer fieldwork costs of approximately £1.48 ha-1. At national scales, £14 million pounds across 4.7 M ha-1 in fieldwork costs per survey could be saved by farmer participation. [ABSTRACT FROM AUTHOR]

The load transfer difficulty caused by borehole wall friction severely limits the penetration rate and extended-reach limit of complex structural wells. A new friction reduction technology termed “earthworm-like drilling” is proposed in this paper to improve the load transfer of complex structural wells. A mathematical model based on a “soft-string” model is developed and solved. The results show that earthworm-like drilling is more effective than single-point vibration drilling. The amplitude and frequency of the pulse pressure and the installation position of the shakers have a substantial impact on friction reduction and load transfer. An optimization model based on the projection gradient method is developed and used to optimize the position of three shakers in a horizontal well. The results verify the feasibility and advantages of earthworm-like drilling, and establish a solid theoretical foundation for its application in oil field drilling. [ABSTRACT FROM AUTHOR]

Background Long-term benthic monitoring is rewarding in terms of science, but labour-intensive, whether in the field, the laboratory, or behind the computer. Building and managing databases require multiple skills, including consistency over time as well as organisation via a systematic approach. Here, we introduce and share our spatially explicit benthic database, comprising 11 years of benthic data. It is the result of intensive benthic sampling that has been conducted on a regular grid (259 stations) covering the intertidal mudflats of the Pertuis-Charentais (Marennes-Oléron Bay and Aiguillon Bay). Samples were taken by foot or by boats during winter depending on tidal height, from December 2003 to February 2014. The present dataset includes abundances and biomass densities of all mollusc species of the study regions and principal polychaetes as well as their length, accessibility to shorebirds, energy content and shell mass when appropriate and available. This database has supported many studies dealing with the spatial distribution of benthic invertebrates and temporal variations in food resources for shorebird species as well as latitudinal comparisons with other databases. In this paper, we introduce our benthos monitoring, share our data, and present a "guide of good practices" for building, cleaning and using it efficiently, providing examples of results with associated R code. New information The dataset has been formatted into a geo-referenced relational database, using PostgreSQL open-source DBMS. We provide density information, measurements, energy content and accessibility of thirteen bivalve, nine gastropod and two polychaete taxa (a total of 66,620 individuals) for 11 consecutive winters. Figures and maps are provided to describe how the dataset was built, cleaned, and how it can be used. This dataset can again support studies concerning spatial and temporal variations in species abundance, interspecific interactions as well as evaluations of the availability of food resources for small- and medium size shorebirds and, potentially, conservation and impact assessment studies. [ABSTRACT FROM AUTHOR]

Many new marine parasite species are added every year. Still, in some places, mainly tropical regions, marine parasites have been little studied. An exhaustive review of the indexed publications where species of parasites are reported in the marine environments of Costa Rica was carried out. The history of research on marine parasites in this region is also reviewed. A total of 147 species of marine parasites have been reported in Costa Rica as parasites of 61 different species of hosts. Most of these parasites correspond to trematodes and cestodes, found mainly in the digestive tract of their vertebrate hosts. In Costa Rica, marine parasites have been studied mainly in sea turtles, elasmobranchs, fish, and dolphins. Most marine parasites have been reported based on morphological identifications of adult stages, and most of the work done so far consists of taxonomic identifications (species reports), with little contribution to the pathology and other aspects of the parasites–hosts interactions. The technical difficulties for research in marine parasitology, the lack of sampling in certain groups of hosts, and the lack of a consolidated research group in marine wildlife parasitology in Costa Rica are factors that have prevented a greater and faster advance in the knowledge of the biodiversity of marine parasites in this country. [ABSTRACT FROM AUTHOR]

Species co-occurrence and site-specific characteristics have a great influence on biotic community composition at local scales and thus contribute to large variations at broad spatial scales. In this paper, we studied invertebrate communities in 63 river sites of the Lower Mekong Basin (LMB) sampled over 609 thousand km 2 . We identified important macroinvertebrate taxa of the component communities (i.e. annelids, crustaceans, mollusks and insects), and key geo-environmental factors that explained the total variance (BD Total ) of the communities at large spatial scale. We used the “ Species Contributions to Beta Diversity” (SCBD) and “ Local Contributions to Beta Diversity” (LCBD) approaches to partition total beta diversity (BD Total ), identified the important macroinvertebrate taxa (those with high SCBD indices), and estimated the uniqueness of sites in community composition (LCBD indices). SCBD indices showed which taxa were the most important in structuring the four component communities: there were 29 insect taxa, which mainly characterized the upstream sites, and 18 mollusk, 7 annelid and 6 crustacean taxa, which all represented the downstream sites. We used linear regression models to investigate the influence of geo-environmental factors and of component communities on LCBD indices. Our results showed great variation in composition within the LMB (BD Total = 0.80 on a 0-to-1 scale). Five sites of the main channel exhibited significant uniqueness (LCBD indices) in community composition. One of them was a hotspot location occupied by a community with exceptional taxonomic composition, which should be protected. Four other sites were degraded by human activity and in need of restoration. Multiple regressions indicated that the global LCBD indices are better explained by the environmental factors, i.e. water conductivity, river depth and Secchi depth (adjusted R 2 = 0.26), than by the geographical factors. Among the component communities, mollusks’ and insects’ LCBDs were the determinants responsible for the variation in the global LCBD indices (adjusted R 2 = 0.84). The uniqueness in community composition of the sites (i.e. LCBDs) that we estimated provides useful ecological information, which could be used to support restoration and conservation planning for the LMB. [ABSTRACT FROM AUTHOR]

Simple Summary: Energy maintenance in living organisms is crucial for survival. The ice-obligate worm, Mesenchytraeus solifugus, displays an unusual bioenergetic pattern, namely that intracellular ATP levels increase with declining temperature. In this study, we address the effects of stress on mitochondrial inorganic polyphosphate (polyP) and its relationship with ATP. Mitochondrial inorganic polyphosphate is a ubiquitous polymer whose role in the maintenance of prokaryotic and mammalian bioenergetics has been broadly demonstrated. We show here that polyP levels in ice worms increase with thermal stress, in contrast with those observed in other annelid worms. Thus, polyP may function as an energetic buffer in ice worms, effectively storing phosphate groups under stress and replenishing ATP under normal physiological conditions. The inorganic polymer, polyphosphate (polyP), is present in all organisms examined to date with putative functions ranging from the maintenance of bioenergetics to stress resilience and protein homeostasis. Bioenergetics in the glacier-obligate, segmented worm, Mesenchytraeus solifugus, is characterized by a paradoxical increase in intracellular ATP levels as temperatures decline. We show here that steady-state, mitochondrial polyP levels vary among species of Annelida, but were elevated only in M. solifugus in response to thermal stress. In contrast, polyP levels decreased with temperature in the mesophilic worm, Enchytraeus crypticus. These results identify fundamentally different bioenergetic strategies between closely related annelid worms, and suggest that I worm mitochondria maintain ATP and polyP in a dynamic equilibrium. [ABSTRACT FROM AUTHOR]

[Display omitted] • Meta-analyses were performed on metallic element accumulation in earthworms. • Metallic element accumulation was significant in earthworms when treated together. • The intensity of body concentration enrichment had a highly species-specific pattern. • Copper uptake in earthworms increased significantly with the increase in soil pH. • Lead accumulation intensity decreased significantly with time in the studied species. The responses of earthworms to excess soil element concentrations are well studied. However, published information on the metallic element accumulation in individuals is controversial. In this paper, the published data on earthworm As, Cd, Cr, Cu, Ni, Pb, and Zn whole body concentrations were evaluated in individuals collected from contaminated and uncontaminated (control) soils, using meta -analyses. The role of soil pH and exposure time as potential influencing factors on metal accumulation was also assessed. Based on the evaluations, the accumulation of each metallic element was significantly (p < 0.05) more intensive in individuals collected from contaminated soils than in ones from control soils, with minor differences in the order of accumulation intensity among the studied metallic elements. Further, major interspecific differences were indicated in the accumulation, with different species being the most intensive accumulators for individual metallic elements. Among the studied metals, Cu concentration in earthworm bodies increased significantly with increasing soil pH. As for the exposure time-dependent accumulation, Pb concentration was found to decrease significantly with time in whole body tissues of earthworms. These results suggested a high variability in metal- and species-specific accumulation-excretion patterns of earthworms, influenced also by other external factors. Based on the results highlighted in this meta -analysis, accumulation schemes raise the need for further analyses involving other additional variables (e.g., soil type, organic matter content, climatic condition) to get a better understanding of element cycle-earthworm relations. [ABSTRACT FROM AUTHOR]

The responses of earthworms to excess soil element concentrations are well studied. However, published information on the metallic element accumulation in individuals is controversial. In this paper, the published data on earthworm As, Cd, Cr, Cu, Ni, Pb, and Zn whole body concentrations were evaluated in individuals collected from contaminated and uncontaminated (control) soils, using meta-analyses. The role of soil pH and exposure time as potential influencing factors on metal accumulation was also assessed. Based on the evaluations, the accumulation of each metallic element was significantly (p

Abstract Background Methylation of cytosines in DNA (5mC methylation) is a major epigenetic modification that modulates gene expression and constitutes the basis for mechanisms regulating multiple aspects of embryonic development and cell reprogramming in vertebrates. In mammals, 5mC methylation of promoter regions is linked to transcriptional repression. Transcription regulation by 5mC methylation notably involves the nucleosome remodeling and deacetylase complex (NuRD complex) which bridges DNA methylation and histone modifications. However, less is known about regulatory mechanisms involving 5mC methylation and their function in non-vertebrate animals. In this paper, we study 5mC methylation in the marine annelid worm Platynereis dumerilii, an emerging evolutionary and developmental biology model capable of regenerating the posterior part of its body post-amputation. Results Using in silico and experimental approaches, we show that P. dumerilii displays a high level of DNA methylation comparable to that of mammalian somatic cells. 5mC methylation in P. dumerilii is dynamic along the life cycle of the animal and markedly decreases at the transition between larval to post-larval stages. We identify a full repertoire of mainly single-copy genes encoding the machinery associated with 5mC methylation or members of the NuRD complex in P. dumerilii and show that this repertoire is close to the one inferred for the last common ancestor of bilaterians. These genes are dynamically expressed during P. dumerilii development and regeneration. Treatment with the DNA hypomethylating agent Decitabine impairs P. dumerilii larval development and regeneration and has long-term effects on post-regenerative growth. Conclusions Our data reveal high levels of 5mC methylation in the annelid P. dumerilii, highlighting that this feature is not specific to vertebrates in the bilaterian clade. Analysis of DNA methylation levels and machinery gene expression during development and regeneration, as well as the use of a chemical inhibitor of DNA methylation, suggest an involvement of 5mC methylation in P. dumerilii development and regeneration. We also present data indicating that P. dumerilii constitutes a promising model to study biological roles and mechanisms of DNA methylation in non-vertebrate bilaterians and to provide new knowledge about evolution of the functions of this key epigenetic modification in bilaterian animals.

Polychaetes belonging to the genus Capitella are known to represent a complex of opportunistic cryptic species that dominates the macrobenthos of organically enriched sediments, such as muddy areas particularly rich in sulfide. With the exception of the species, Capitella capitata (Fabricius) from West Greenland, Capitella spp. from the European Atlantic coast have never been accurately characterized both genetically and morphologically and were often reported as capitata in the literature. The life cycle of these European worms has not been described properly either, despite their widespread use as bio-indicators in ecological studies. The present study provides the first morphological description of Capitella specimens collected along the Brittany coast and the English Channel up to the entrance of the North Sea and their genetic diversification in this region. Both morphological and molecular data are congruent and support the co-occurrence of cryptic species at the tip of Brittany. The most frequent French mt lineages, C-Channel1, C-Atlantic and C-Channel2, although well distinct from Capitella teleta, are also divergent but closer to C. capitata initially described from Greenland. Following barcoding, the most abundant species (C-Channel1) was isolated and reared in the laboratory to describe its life cycle and predict both its dispersal ability and ecological success in the face of the sulfidic muddy habitats of the French harbors. [ABSTRACT FROM AUTHOR]

We present an updated checklist of marine annelids from the Indian subcontinent. Records of annelid species were obtained from published and grey literatures and online databases. Our review of annelid publications was restricted to the Indian continental shelf region. This paper also discusses the taxonomic status of marine annelid species recorded from this region and problems that impede its research. The updated list comprises of 727 species belonging to 334 genera and 72 families. A total of 152 species have their type locality in India including 88 species that are endemic to the region. The current checklist indicates that 25% (183 species) of the records are questionable and require further examination. Our results highlight that marine annelid richness of the Indian sub-continent is underestimated with many of the native undescribed species being most likely concealed under ‘erroneous’ or ‘cosmopolitan’ records. With a combination of factors that include a lack of experts, funding, and failure to update regional literature has resulted in an incomplete state of knowledge for the marine annelid biodiversity from this region. Therefore, there is an urgent need for extensive and intensive sampling to discover new species, conduct detailed re-examinations of doubtful records and, collaborate within the local and international institutes and organizations to improve the regional biodiversity studies.

The oligochaete annelids, belonging to the subfamily Tubificinae, produce two types of spermatozoa: eusperm (the fertilising ones) and parasperm, protecting and carrying the eusperm. The pathway for the production of the two types is common until the onset of meiosis, but then a regular meiosis produces eusperm, whereas parasperm is generated through a peculiar mechanism of nuclear fragmentation giving rise to an irregular, but very high, number of paraspermatozoa. Since every parasperm has its own flagellum, this entails the necessity of producing a very high number of basal bodies. In the present paper, we describe how basal bodies are generated through a mechanism similar to that producing the basal bodies in ciliated epithelia, but never observed up to now during the genesis of a uniflagellated cell. Basal bodies form in close proximity to a precursor structure called deuterosome, which originates de novo in the cytoplasm from fibrogranular material. The various stages of centriologenesis are positive to anti-centrin antibodies and, observed by electron microscopy, correspond closely to the ones described for ciliated epithelia. However, once formed, the basal bodies migrate to their final position and produce the parasperm flagellum. [Copyright &y& Elsevier]

Based on collections of interstitial Syllidae (Annelida) from several areas of the Indo-Pacific Ocean (Madagascar, Andaman Islands, South China, Hong-Kong, the Philippines, and New Zealand), we report a total of 49 species belonging to 19 genera. Most of these species are already known in the Indo-Pacific area, but these reports extend their distributional ranges. Nine species are described as new: Parexogone javieri (from New Zealand), Parexogone viejoi (from the Philippines), Prosphaerosyllis fittoni, Sphaerosyllis dieteri, Syllis kai, Perkinsyllis tsilo (from Madagascar), and Megasyllis chiki, Syllis dominguezi, and Syllis escribanoi (from China). Two species, previously described as StreptosyllisWebster & Benedict, 1884 are transferred to the genus StreptospinigeraKudenov, 1983: S. baolingi (Ding & Westheide, 1994), new combination, and S. hainanensis(Ding & Westheide, 1994), new combination, both from China. One species is transferred to Syllis [Typosyllis botosaneanui (Hartmann-Schröder, 1973)], new combination. [ABSTRACT FROM AUTHOR]

Recent studies have shown that, despite stereotypical cleavage, annelids show the ability for embryonic regulation, including the formation of germline cells. However, the widest variety of regulative processes is presented in the postlarval development of annelids. The ability to regenerate, which is probably an ancestral feature, manifests itself variously among these animals. Some species are unable to regenerate lost segments. However, most species replace lost posterior body parts, many are able to reestablish missing head segments and structures, and some develop the entire body de novo even on the basis of one or two segments. Most of the regenerated structures are formed due to a set of undifferentiated cells arising from the division of dedifferentiated and/or stem cells. Moreover, the regeneration process often involves remodeling of survived body fragments and may thus be associated not only with local changes but also require a response at the level of the whole organism. In this review, we summarize many recent studies on the molecular and cellular mechanisms of regeneration in annelids. Special attention is paid to the regeneration of the digestive and nervous systems and integuments as well as to the involvement of stem and undifferentiated cells in the development of blastema and in replacing the lost gonads. Accumulation and analysis of recent findings about the diversity of cellular sources and mechanisms of annelid regeneration may shed light on the most evolutionarily conserved programs for maintaining regeneration ability and processes leading to the loss (limitation) of one of the ancestral features of animals. [ABSTRACT FROM AUTHOR]

The presence of earthworm species in crop fields is as old as agriculture itself. The earthworms Pontoscolex corethrurus (invasive) and Balanteodrilus pearsei (native) are associated with the emergence of agriculture and sedentism in the region Amazon and Maya, respectively. Both species have shifted their preference from their natural habitat to the cropland niche. They contrast in terms of intensification of agricultural land use (anthropic impact to the symbiotic soil microbiome). P. corethrurus inhabits conventional agroecosystems, while B. pearsei thrives in traditional agroecosystems, i.e., P. corethrurus has not yet been recorded in soils where B. pearsei dwells. The demographic behavior of these two earthworm species was assessed in the laboratory over 100 days, according to their origin (OE; P. corethrurus and B. pearsei) food quality (FQ; soil only, maize stubble, Mucuna pruriens), and soil moisture (SM; 25, 33, 42%). The results showed that OE, FQ, SM, and the OE x FQ interaction were highly significant for the survival, growth, and reproduction of earthworms. P. corethrurus showed a lower survival rate (> mortality). P. corethrurus survivors fed a diet of low-to-intermediate nutritional quality (soil and stubble maize, respectively) showed a greater capacity to grow and reproduce; however, it was surpassed by the native earthworm when fed a high-quality diet (M. pruriens). Besides, P. corethrurus displayed a low cocoon hatching (emergence of juveniles). These results suggest that the presence of the invasive species was associated with a negative interaction with the soil microbiota where the native species dwells, and with the absence of natural mutualistic bacteria (gut, nephridia, and cocoons). These results are consistent with the absence of P. corethrurus in milpa and pasture-type agricultural niches managed by peasants (agroecologists) to grow food regularly through biological soil management. Results reported here suggest that P. corethrurus is an invasive species that is neither wild nor domesticated, that is, its eco-evolutionary phylogeny needs to be derived based on its symbionts. [ABSTRACT FROM AUTHOR]

An antifungal active fraction (AAF) from the coelomic fluid (CF) of the earthworm Dendrobaena veneta was isolated. The aim of the study was to analyze the antifungal activity of the AAF and to carry out chemical characterization of the fraction. The active fraction showed antifungal activity against a clinical C. albicans isolate, C. albicans ATCC 10231, and C. krusei ATCC 6258. It effectively reduced the metabolic activity of C. albicans cells and influenced their morphology after 48 hours of incubation. Scanning electron microscopy (SEM) images revealed loss of integrity of the cell wall induced by the active fraction. Calcofluor White staining showed changes in the structure of the C. albicans cell wall induced by the AAF. The fungal cells died via apoptosis and necrosis after the treatment with the studied fraction. Electrophoresis under native conditions revealed the presence of two compounds in the AAF, while SDS/PAGE gel electrophoresis showed several protein and carbohydrate compounds. The active fraction was analyzed using Raman spectroscopy, MALDI TOF/TOF, and ESI LC-MS. The Raman analysis confirmed the presence of proteins and determined their secondary structure. The MALDI TOF/TOF analysis facilitated detection of four main compounds with a mass of 7694.9 m/z, 12292.3 m/z, 21628.3 m/z, and 42923.2 m/z in the analyzed fraction. The presence of carbohydrate compounds in the preparation was confirmed by nuclear magnetic resonance (NMR) and gas chromatography (GC-MS). The ATR-FTIR spectrum of the AAF exhibited high similarity to the spectrum of egg white lysozyme. The AAF showed no endotoxicity and cytotoxicity towards normal skin fibroblasts (HSF); therefore, it can be used for the treatment of skin and mucous membrane candidiasis in the future. Given its efficient and selective action, the fraction seems to be a promising preparation with antifungal activity against C. albicans. [ABSTRACT FROM AUTHOR]

Lumbricidae taxonomy is vastly restricted by the morphological simplicity of earthworms and their lack of complex appendices. This has led to confusing results in the Lumbricidae classifications, which in turn, has hindered our ability to identify and assign new and cryptic species to the family. Here we propose the addition of a new Lumbricidae genus from the Zagros and Elburz Mountains of Iran, i.e. Philomontanus gen. nov, including three new species. Our taxonomic inferences were based on the phylogenetic analysis of two nuclear gene regions (28S rDNA and 18S rDNA) and 11 mitochondrial gene regions (16S rDNA, 12S rDNA, NADH dehydrogenase I, cytochrome oxidase subunits I and II and tRNAs Asn, Asp, Val, Leu, Ala and Ser). Philomontanus gen. nov comprises the earthworm species Philomontanus sarii sp. nov., Philomontanus mahmoudi sp. nov. and Philomontanus baloutchi sp. nov. These three species are morphologically similar to each other with only a few characters separating them (e.g. size, pigmentation and position of clitellum). Our findings support the adoption of an integrative approach including molecular information (e.g., DNA sequences) to aid earthworm classification and develop a robust taxonomy. [ABSTRACT FROM AUTHOR]

Tracing the evolution of the siboglinid group, peculiar group of marine gutless annelids, requires the detailed study of the fragmentarily explored central nervous system of vestimentiferans and other siboglinids. 3D reconstructions of the neuroanatomy of Riftia revealed that the “brain” of adult vestimentiferans is a fusion product of the supraesophageal and subesophageal ganglia. The supraesophageal ganglion-like area contains the following neural structures that are homologous to the annelid elements: the peripheral perikarya of the brain lobes, two main transverse commissures, mushroom-like structures, commissural cell cluster, and the circumesophageal connectives with two roots which give rise to the palp neurites. Three pairs of giant perikarya are located in the supraesophageal ganglion, giving rise to the paired giant axons. The circumesophageal connectives run to the VNC. The subesophageal ganglion-like area contains a tripartite ventral aggregation of perikarya (= the postoral ganglion of the VNC) interconnected by the subenteral commissure. The paired VNC is intraepidermal, not ganglionated over most of its length, associated with the ciliary field, and comprises the giant axons. The pairs of VNC and the giant axons fuse posteriorly. Within siboglinids, the vestimentiferans are distinguished by a large and considerably differentiated brain. This reflects the derived development of the tentacle crown. The tentacles of vestimentiferans are homologous to the annelid palps based on their innervation from the dorsal and ventral roots of the circumesophageal connectives. Neuroanatomy of the vestimentiferan brains is close to the brains of Cirratuliiformia and Spionida/Sabellida, which have several transverse commissures, specific position of the giant somata (if any), and palp nerve roots (if any). The palps and palp neurite roots originally developed in all main annelid clades (basally branching, errantian and sedentarian annelids), show the greatest diversity in their number in sedentarian species. Over the course of evolution of Sedentaria, the number of palps and their nerve roots either dramatically increased (as in vestimentiferan siboglinids) or were lost. [ABSTRACT FROM AUTHOR]

This research was conducted to compare chemical and microbiological properties during aerobic composting (AC) and vermicomposting (VC) of green waste. Relative to AC, VC significantly decreased the pH and lignin and cellulose contents, and significantly increased the electrical conductivity and total N and available P contents. For AC, BIrii41_norank (order Myxococcales) was the major bacterial genus at 30 d and again became dominant genus from 90–150 d, with relative abundances of 2.88% and 4.77–5.19%, respectively; at 45 d and 60 d, the dominant bacterial genus was Nitrosomonadaceae_uncultured (order Nitrosomonadales) with relative abundances of 2.83–7.17%. For VC, the dominant bacterial genus was BIrii41_norank (except at 45 d), which accounted for 2.11–7.96% of the total reads. The dominant fungal class was Sordariomycetes in AC (relative abundances 39.2–80.6%) and VC (relative abundances 42.1–69.5%). The abundances of microbial taxa and therefore the bacterial and fungal community structures differed between VC and AC. The quality of the green waste compost product was higher with VC than with AC. These results will also help to achieve further composting technology breakthroughs in reducing the composting time and improving compost quality. [ABSTRACT FROM AUTHOR]

Uniformly pigmented Eisenia andrei (Ea) and striped E. fetida (Ef) lumbricid earthworms are hermaphrodites capable of self-fertilization, cross-fertilization, and asymmetrical hybridization. The latter was detected by genotyping of F1 and F2 progeny of the controlled Ea+Ef pairs by species-specific sequences of maternal mitochondrial COI genes and maternal/paternal nuclear S28 rRNA genes. Among F1offspring there were self-fertilized Ea (aAA), Ef (fFF), and cross-fertilized fertile Ea-derived hybrids (aAF); the latter mated with Ea and gave new generation of Ea and hybrids, while mated with Ef gave Ea, Ef, Ea-derived hybrids and sterile Ef-derived hybrids (fFA). Coelomic fluid of Ea exhibits unique fluorescence spectra called here the M-fluorescence considered as a molecular biomarker of this species. Since similar fluorescence was detected also in some Ef (hypothetical hybrids?), the aim of present investigations was to identify the M-positive earthworms among families genotyped previously. It was assumed that factor/s responsible for metabolic pathways leading to production of undefined yet M-fluorophore might be encoded/controlled by alleles of hypothetical nuclear gene of Eisenia sp. segregating independently from species-specific S28 rRNA nuclear genes, where ‘MM’ or ‘Mm’ alleles determine M-positivity while ‘mm’ alleles determine M-negative phenotypes. Spectra of M-fluorescence were detected in all 10 Ea (aAAMM) and 19 Ea-derived hybrids (aAFMm), three of four Ef-derived hybrids (fFAMm) and one ‘atypical’ Ef (fFFMm) among 13 Ef earthworms. Among progeny of ‘atypical’ M-positive Ef (fFFMm) reappeared ‘typical’ M-negative Ef (fFFmm), confirming such hypothesis. Alternatively, the M-fluorescence might be dependent on unknown gene products of vertically-transmitted Ea-specific symbiotic bacteria sexually transferred to the Ef partner. Hypotheses of intrinsic and external origin of M-fluorescence might complement each other. The presence/absence of M-fluorophore does not correspond with body pigmentation patterns; Ef-characteristic banding appeared in posterior parts of hybrids body. In conclusion, Ea/Ef hybridization may serve for further studies on bi-directional gene flow. [ABSTRACT FROM AUTHOR]

The medicinal leech is one of the most venerated model systems for the study of fundamental nervous system principles, ranging from single-cell excitability to complex sensorimotor integration. Yet, molecular analyses have yet to be extensively applied to complement the rich history of electrophysiological study that this animal has received. Here, we generated the first de novo transcriptome assembly from the entire central nervous system of Hirudo verbana, with the goal of providing a molecular resource, as well as to lay the foundation for a comprehensive discovery of genes fundamentally important for neural function. Our assembly generated 107,704 contigs from over 900 million raw reads. Of these 107,704 contigs, 39,047 (36%) were annotated using NCBI’s validated RefSeq sequence database. From this annotated central nervous system transcriptome, we began the process of curating genes related to nervous system function by identifying and characterizing 126 unique ion channel, receptor, transporter, and enzyme contigs. Additionally, we generated sequence counts to estimate the relative abundance of each identified ion channel and receptor contig in the transcriptome through Kallisto mapping. This transcriptome will serve as a valuable community resource for studies investigating the molecular underpinnings of neural function in leech and provide a reference for comparative analyses. [ABSTRACT FROM AUTHOR]

Heterosporis sutherlandae is an emerging microsporidian fish parasite in the Great Lakes region. H. sutherlandae forms lesions in the muscle tissue of fishes important to aquaculture and sport fishing. These lesions render the filet inedible and may have fitness consequences. We evaluated the prevalence and severity of H. sutherlandae among yellow perch (Perca flavescens) in a known-positive Minnesota lake, and used an equilibrium yield model to evaluate impacts on harvest. Twenty-eight percent of the 400 yellow perch sampled were infected with H. sutherlandae. Males were 1.5 times more likely to be infected than females and were more severely infected. The presence of the parasite did not vary with relative weight or age, but infection severity was highest among older individuals that were in better condition. These results suggest that males are more susceptible to infection, and that infection is not associated with maturity or a gape-limiting food source. These results also suggest that heterosporosis increases in severity with time or by increased exposure. Our equilibrium yield model found that a 10% increase in mortality due to H. sutherlandae could result in 30% and 10% reductions in yield and mean catch weight, respectively. The results of this study direct future field sampling and laboratory experiments to further understand and predict the impacts of this parasite. [ABSTRACT FROM AUTHOR]

Lumbricid earthworms Eisenia andrei (Ea) and E. fetida (Ef) are simultaneous hermaphrodites with reciprocal insemination capable of self-fertilization while the existence of hybridization of these two species was still debatable. During the present investigation fertile hybrids of Ea and Ef were detected. Virgin specimens of Ea and Ef were laboratory crossed (Ea+Ef) and their progeny was doubly identified. 1 –identified by species-specific maternally derived haploid mitochondrial DNA sequences of the COI gene being either ‘a’ for worms hatched from Ea ova or ‘f’ for worms hatched from Ef ova. 2 –identified by the diploid maternal/paternal nuclear DNA sequences of 28s rRNA gene being either ‘AA’ for Ea, ‘FF’ for Ef, or AF/FA for their hybrids derived either from the ‘aA’ or ‘fF’ ova, respectively. Among offspring of Ea+Ef pairs in F1 generation there were mainly aAA and fFF earthworms resulted from the facilitated self-fertilization and some aAF hybrids from aA ova but none fFA hybrids from fF ova. In F2 generation resulting from aAF hybrids mated with aAA a new generations of aAA and aAF hybrids were noticed, while aAF hybrids mated with fFF gave fFF and both aAF and fFA hybrids. Hybrids intercrossed together produced plenty of cocoons but no hatchlings independently whether aAF+aAF or aAF+fFA were mated. These results indicated that Ea and Ef species, easy to maintain in laboratory and commonly used as convenient models in biomedicine and ecotoxicology, may also serve in studies on molecular basis of interspecific barriers and mechanisms of introgression and speciation. Hypothetically, their asymmetrical hybridization can be modified by some external factors. [ABSTRACT FROM AUTHOR]

Toxic effects of cadmium (Cd), a common soil pollutant, are still not very well understood, particularly in regard to its epigenetic impact. Therefore, the aim of this study was to assess DNA methylation changes and their persistence in the earthworm Lumbricus terrestris upon chronic low dose Cd exposure using methylation sensitive amplification polymorphism (MSAP). Moreover, the biomarker response and fitness of the earthworms, as well as the expression of detoxification-related genes (metallothionein (MT) and phytochelatin synthase (PCS)) was evaluated. Low levels of Cd caused an increase in genome-wide DNA methylation, which remained partly modified, even after several months of recovery in unpolluted soil. Increased cellular stress seemed to decrease after two weeks of exposure whereas fitness parameters remained unaffected by Cd, probably as a result from the activation of detoxification mechanisms like the expression of MTs. Interestingly, even though the level of Cd exposure was very low, MT expression levels indicate the development of acclimation mechanisms. Taken together, this study demonstrates that acclimation, as well as epigenetic modifications can occur already in moderately polluted environments. In addition, these effects can have long-lasting impacts on key species of soil invertebrates and might persist long after the actual heavy metal challenge has passed. [ABSTRACT FROM AUTHOR]

TCTP (Translationally Controlled Tumour Protein) is a multifunctional protein that plays a role in the development, immune system, tumour reversion, and maintenance of stem cells. The mRNA of the Tpt1 gene is over-expressed during liver regeneration. But, the function of the protein in regeneration is not known. To study the role of the protein in regeneration, the earthworm Eudrilus eugeniae was chosen. First, the full length cDNA of the Tpt1 gene was sequenced. The size of the cDNA is 504 bp and the protein has 167 amino acids. The highest level of TCTP expression was documented in the worm after three days of regeneration. The protein was found to be expressed specifically in the epithelial layer of the skin. During regeneration, the protein expression was found to be the highest at the tip of blastema. The pharmacological suppression of TCTP using nutlin-3 and TCTP RNAi experiments resulted in the failure of the regeneration process. The suppression of TCTP caused the arrest of proliferation in posterior amputated worms. The severe cell death was documented in the amputated region of nutlin-3 injected worm. The silencing of TCTP has blocked the modification of clitellar segments. The experiments confirm that TCTP has major functions in the upstream signalling of cell proliferation in the early regeneration process in E. eugeniae. [ABSTRACT FROM AUTHOR]

Gene expression is highly plastic, which can help organisms to both acclimate and adapt to changing environments. Possible variation in gene expression among individuals with the same genotype (among clones) is not widely considered, even though it could impact the results of studies that focus on gene expression phenotypes, for example studies using clonal lines. We examined the extent of within and between clone variation in gene expression in the earthworm Dendrobaena octaedra, which reproduces through apomictic parthenogenesis. Five microsatellite markers were developed and used to confirm that offspring are genetic clones of their parent. After that, expression of 12 genes was measured from five individuals each from six clonal lines after exposure to copper contaminated soil. Variation in gene expression was higher over all genotypes than within genotypes, as initially assumed. A subset of the genes was also examined in the offspring of exposed individuals in two of the clonal lines. In this case, variation in gene expression within genotypes was as high as that observed over all genotypes. One gene in particular (chymotrypsin inhibitor) also showed significant differences in the expression levels among genetically identical individuals. Gene expression can vary considerably, and the extent of variation may depend on the genotypes and genes studied. Ensuring a large sample, with many different genotypes, is critical in studies comparing gene expression phenotypes. Researchers should be especially cautious inferring gene expression phenotypes when using only a single clonal or inbred line, since the results might be specific to only certain genotypes. [ABSTRACT FROM AUTHOR]

Populations of Limnodrilus sulphurensis Fend, Liu & Erséus, 2016 (Naididae, Oligochaeta) were found on sulphur bacterial mats in a stream with sulphidic and hypoxic water in Sulphur Cave, Colorado (USA). In order to reveal adaptations to the hostile conditions, we assessed physical and chemical factors in reference to anatomical and physiological details of L. sulphurensis and compared with the common L. hoffmeisteri from non-sulphidic waters. Sections and 3D reconstructions of the posterior body of L. sulphurensis showed an extensively ramified integumental blood system and an extraordinarily rich circumintestinal chloragogue tissue. This contained accumulations of sulphide and iron. These features coincided with an unusual blood physiology of these haemoglobin-containing annelids living under permanent sulphide exposure. The blood of L. sulphurensis has a high oxygen binding capacity and the binding sites seem unaffected by sulphide (sulphhaemoglobin is not formed). Exposure experiments indicated oxidation of sulphide to thiosulphate, which can be easily released. H plc analysis of the chloragosomes showed the presence of haemin (presumably haematin), a known catalyst of sulphide oxidation. L. sulphurensis shares these features with many marine 'sulfide-annelids'. For the freshwater realm these data are novel. The proposed 'sulfur-haeme scenario' would explain the unrivalled existence of L. sulphurensis under sulphidic conditions. [ABSTRACT FROM AUTHOR]

Di-(2-ethylhexyl) phthalate (DEHP) is a dominant phthalic acid ester (PAE) that has aroused public concern due to its resistance to degradation and its toxicity as an endocrine-disrupting compound. Effects of different concentrations of DEHP on Eisenia fetida in spiked natural soil have been studied in the body of the earthworm by means of soil cultivation tests 7, 14, 21 and 28 days after exposure. The results indicated that, in general, superoxide dismutase (SOD) activity, malondialdehyde (MDA) content, metallothionein (MT) content, the expression of heat shock protein 70 (HSP 70) and all the tested geno-toxicity parameters are promoted as time elapses and with increasing concentration of DEHP. However, peroxidase (POD) activity, neutral red retention time (NRRT) and mitochondrial membrane potential difference values were found to decrease even at a low concentration of DEHP of 1 mg kg-1 soil (p<0.05). Clear toxic effects of DEHP on E. fetida have been generally recognized by means of the disturbance of antioxidant enzyme activity/content and critical proteins, cell membrane and organelle disorder and DNA damage estimated by length of tail, tail DNA ratio, and tail moment parameters. A concentration of DEHP of 3 mg kg-1 may be recommended as a precaution against the potential risk of PAEs in soils and for indicating suitable threshold values for other soil animals and soil micro-organisms. [ABSTRACT FROM AUTHOR]

As a standard testing organism in soil ecosystems, the earthworm Eisenia fetida has been used widely in toxicity studies. However, tests at the individual level are time- and animal-consuming, with limited sensitivity. Earthworm coelomocytes are important for the assimilation and elimination of exogenous compounds and play a key role in the processes of phagocytosis and inflammation. In this study, we explored an optimal condition to culture coelomocytes of E. fetida in vitro and investigated the cytotoxicity of multiwalled carbon nanotubes (MWCNTs) and sodium pentachlorophenol (PCP-Na) using coelomocytes via evaluating lethal toxicity, oxidative stress, membrane damage, and DNA damage. The results showed that coelomocytes can be successfully cultured in vitro in primary under the RPMI-1640 medium with 2–4×104 cells/well (1–2×105 cells/mL) in 96-well plates at 25°C without CO2. Both MWCNTs and PCP-Na could cause oxidative damage and produce ROS, an evidence for lipid peroxidation with MDA generation and SOD and CAT activity inhibition at high stress. The two chemicals could separately damage the cell membrane structure, increasing permeability and inhibiting mitochondrial membrane potential (MMP). In addition, our results indicate that PCP-Na may be adsorbed onto MWCNTs and its toxicity on earthworm was accordingly alleviated, while a synergetic effect was revealed when PCP-Na and MWCNTs were added separately. In summary, coelomocyte toxicity in in vitro analysis is a sensitive method for detecting the adverse effects of carbon nanotubes combined with various pollutants. [ABSTRACT FROM AUTHOR]

The effects of adding the biosurfactant rhamnolipid, the lignolytic and cellulolytic fungus Phanerochete chrysosporium, and the free-living nitrogen-fixing bacterium Azotobacter chrococcum on vermicomposting of green waste with Eisenia fetida was investigated. The addition of rhamnolipid and/or either microorganism alone or in all combinations significantly increased E. fetida growth rate, the number of E. fetida juveniles and cocoons, the population densities of cellulolytic fungi and Azotobacter bacteria, and cellulase and urease activities in the vermicomposts. The quality of the final vermicompost (in terms of electrical conductivity, nutrient content, C/N ratio, humic acid content, lignin and cellulose contents, and phytotoxicity to germinating seeds) was enhanced by addition of rhamnolipid and/or microorganisms. The physical characteristics of vermicomposts produced with rhamnolipid and/or microorganisms were acceptable for agricultural application. The best quality vermicompost was obtained with the combined addition of P. chrysosporium, A. chrococcum, and rhamnolipid. [ABSTRACT FROM AUTHOR]

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Exceptionally preserved 'Burgess Shale-type' fossil assemblages from the Cambrian of Laurentia, South China and Australia record a diverse array of non-biomineralizing organisms. During this time, the palaeocontinent Baltica was geographically isolated from these regions, and is conspicuously lacking in terms of comparable accessible early Cambrian Lagerstätten. Here we report a diverse assemblage of small carbonaceous fossils ( SCFs) from the early Cambrian (Stage 4) File Haidar Formation of southeast Sweden and surrounding areas of the Baltoscandian Basin, including exceptionally preserved remains of Burgess Shale-type metazoans and other organisms. Recovered SCFs include taxonomically resolvable ecdysozoan elements (priapulid and palaeoscolecid worms), lophotrochozoan elements (annelid chaetae and wiwaxiid sclerites), as well as 'protoconodonts', denticulate feeding structures, and a background of filamentous and spheroidal microbes. The annelids, wiwaxiids and priapulids are the first recorded from the Cambrian of Baltica. The File Haidar SCF assemblage is broadly comparable to those recovered from Cambrian basins in Laurentia and South China, though differences at lower taxonomic levels point to possible environmental or palaeogeographical controls on taxon ranges. These data reveal a fundamentally expanded picture of early Cambrian diversity on Baltica, and provide key insights into high-latitude Cambrian faunas and patterns of SCF preservation. We establish three new taxa based on large populations of distinctive SCFs: Baltiscalida njorda gen. et sp. nov. (a priapulid), Baltichaeta jormunganda gen. et sp. nov. (an annelid) and Baltinema rana gen. et sp. nov. (a filamentous problematicum). [ABSTRACT FROM AUTHOR]

Mixing soil or adding earthworms (Eisenia fetida (, 1826)) accelerated the removal of anthracene, a polycyclic aromatic hydrocarbon, from a pasture and an arable soil, while a non-ionic surfactant (Surfynol® 485) inhibited the removal of the contaminant compared to the untreated soil. It was unclear if the treatments affected the soil bacterial community and consequently the removal of anthracene. Therefore, the bacterial community structure was monitored by means of 454 pyrosequencing of the 16S rRNA gene in the pasture and arable soil mixed weekly, amended with Surfynol® 485, E. fetida or organic material that served as food for the earthworms for 56 days. In both soils, the removal of anthracene was in the order: mixing soil weekly (100%) > earthworms applied (92%) > organic material applied (77%) > untreated soil (57%) > surfactant applied (34%) after 56 days. There was no clear link between removal of anthracene from soil and changes in the bacterial community structure. On the one hand, application of earthworms removed most of the contaminant from the arable soil and had a strong effect on the bacterial community structure, i.e. a decrease in the relative abundance of the Acidobacteria, Chloroflexi and Gemmatimonadetes, and an increase in that of the Proteobacteria compared to the unamended soil. Mixing the soil weekly removed all anthracene from the arable soil, but had little or no effect on the bacterial community structure. On the other hand, application of the surfactant inhibited the removal of anthracene from the arable soil compared to the untreated soil, but had a strong effect on the bacterial community structure, i.e. a decrease in the relative abundance of Cytophagia (Bacteroidetes), Chloroflexi, Gemmatimonadetes and Planctomycetes and an increase in that of the Flavobacteria (Bacteroidetes) and Proteobacteria. Additionally, the removal of anthracene was similar in the different treatments of both the arable and pasture soil, but the effect of application of carrot residue, earthworms or the surfactant on the bacterial community structure was more accentuated in the arable soil than in the pasture soil. It was found that removal of anthracene was not linked to changes in the bacterial community structure. [ABSTRACT FROM AUTHOR]

Gene encoded antimicrobial peptides (AMPs) are widely distributed among living organisms including plants, invertebrates and vertebrates. They constitute important effectors of the innate immune response by exerting multiple roles as mediators of inflammation with impact on epithelial and inflammatory cells influencing diverse processes such as cytokine release, cell proliferation, angiogenesis, wound healing, chemotaxis and immune induction. In invertebrates, most of the data describe the characterization and/or the function of AMPs in the numerically and economically most representative group which are arthropods. Annelids are among the first coelomates and are therefore of special phylogenetic interest. Compared to other invertebrate groups, data on annelid’s immunity reveal heavier emphasis on the cellular than on the humoral response suggesting that immune defense of annelids seems to be principally developed as cellular immunity.This paper gives an overview of the variety of AMPs identified in the three classes of annelids, i.e. polychaetes, oligochaetes and achaetes. Their functions, when they have been studied, in the humoral or cellular response of annelids are also mentioned.

Members of the skipper tribe Baorini generally resemble each other and are characterized by dark brown wings with hyaline white spots. These shared characteristics have caused difficulties with revealing the relationships among genera and species in the group, and some conflicting taxonomic views remain unresolved. The present study aims to infer a more comprehensive phylogeny of the tribe using molecular data, to test the monophyly of the tribe as well as the genera it includes in order to clarify their taxonomic status, and finally to revise the current classification of the group. In order to reconstruct a phylogenetic tree, the mitochondrial COI-COII and 16S genes as well as the nuclear EF-1α and 28S genes were analyzed using parsimony, maximum likelihood, and Bayesian inference. The analysis included 67 specimens of 41 species, and we confirmed the monophyly of Baorini, and revealed that 14 genera are well supported. The genus Borbo is separated into three clades: Borbo, Pseudoborbo, and Larsenia gen. nov. We confirmed that Polytremis is polyphyletic and separated into three genera: Polytremis, Zinaida, and Zenonoida gen. nov., and also confirmed that the genus Prusiana is a member of the tribe. Relationships among some genera were strongly supported. For example, Zenonia and Zenonoida were found to be sister taxa, closely related to Zinaida and Iton, while Pelopidas and Baoris were also found to cluster together. [ABSTRACT FROM AUTHOR]

Elucidating the basic life-history of endangered species is the first important step in the conservation of such species. This study examined the reproductive ecology and the preimaginal morphology of the endangered ground beetle Elaphrus sugai Nakane (Coleoptera: Carabidae); currently, the Watarase wetland of the central Kanto Plain, Japan is the only confirmed locality of this beetle species. Laboratory rearing of reproductive adults collected in early April revealed that females can lay more than 131 eggs. Eggs were laid in mud, without an egg chamber. Larvae reached adulthood when fed a diet of mealworms, indicating that E. sugai larvae are insect larvae feeders. An earthworm diet, the optimal diet for larvae of a congeneric species (E. punctatus Motschulsky), was lethal to E. sugai larvae. The egg stage was 3–4 days in duration under a 16L8D cycle (22°C). The duration from hatching to adult eclosion was 23–42 days at various temperatures simulating those of the reproductive period. Larval morphology was similar to that of consubgeneric species described previously. The pupa is unusual, in that the setae on the abdominal tergites are long (twice as long as those of the abdominal segment) and have somewhat “coiled” apices. Finally, the current endangered status of E. sugai was compared to that of E. viridis Horn, which has been regarded as the most endangered species of the genus worldwide. [ABSTRACT FROM AUTHOR]

Formation of extracellular traps (ETs) capturing and immobilizing pathogens is now a well-established defense mechanism added to the repertoire of vertebrate phagocytes. These ETs are composed of extracellular DNA (extDNA), histones and antimicrobial proteins. Formation of mouse and human ETs depends on enzymes (i) facilitating decondensation of chromatin by citrullination of histones, and (ii) serine proteases degrading histones. In invertebrates, initial reports revealed existence of ETs composed of extDNA and histones, and here we document for the first time that also coelomocytes, immunocompetent cells of an earthworm Eisenia andrei, cast ETs which successfully trap bacteria in a reactive oxygen species (ROS)-dependent and -independent manner. Importantly, the formation of ETs was observed not only when coelomocytes were studied ex vivo, but also in vivo, directly in the earthworm coelom. These ETs were composed of extDNA, heat shock proteins (HSP27) and H3 histones. Furthermore, the formation of E. andrei ETs depended on activity of serine proteases, including elastase-like activity. Moreover, ETs interconnected and hold together aggregating coelomocytes, a processes proceeding encapsulation. In conclusion, the study confirms ET formation by earthworms, and unravels mechanisms leading to ET formation and encapsulation in invertebrates. [ABSTRACT FROM AUTHOR]

Background: Earthworms are important for maintaining soil ecosystem functioning and serve as indicators of soil fertility. However, detection of earthworms is time-consuming, which hinders the assessment of earthworm abundances with high sampling density over entire fields. Recent developments of mobile terrestrial sensor platforms for proximal soil sensing (PSS) provided new tools for collecting dense spatial information of soils using various sensing principles. Yet, the potential of PSS for assessing earthworm habitats is largely unexplored. This study investigates whether PSS data contribute to the spatial prediction of earthworm abundances in species distribution models of agricultural soils. Methodology/Principal Findings: Proximal soil sensing data, e.g., soil electrical conductivity (EC), pH, and near infrared absorbance (NIR), were collected in real-time in a field with two management strategies (reduced tillage / conventional tillage) and sandy to loam soils. PSS was related to observations from a long-term (11 years) earthworm observation study conducted at 42 plots. Earthworms were sampled from 0.5 x 0.5 x 0.2 m³ soil blocks and identified to species level. Sensor data were highly correlated with earthworm abundances observed in reduced tillage but less correlated with earthworm abundances observed in conventional tillage. This may indicate that management influences the sensor-earthworm relationship. Generalized additive models and state-space models showed that modelling based on data fusion from EC, pH, and NIR sensors produced better results than modelling without sensor data or data from just a single sensor. Regarding the individual earthworm species, particular sensor combinations were more appropriate than others due to the different habitat requirements of the earthworms. Earthworm species with soil-specific habitat preferences were spatially predicted with higher accuracy by PSS than more ubiquitous species. Conclusions/Significance: Our findings suggest that PSS contributes to the spatial modelling of earthworm abundances at field scale and that it will support species distribution modelling in the attempt to understand the soil-earthworm relationships in agroecosystems. [ABSTRACT FROM AUTHOR]