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Identifying the earthworm material recently collected in different parts of Turkey (Marmara Region, Upper Mesopotamia, Hatay Province and East Anatolia) resulted in records of 29 earthworm species including three lumbricids new to science: Dendrobaena cevdeti, D. szalokii and Eisenia patriciae spp. nov. In addition, Dendrobaena cognettii is reported for the first time from the country. With this contribution, the number of earthworm species and subspecies registered in Turkey is raised to 80.

The Island of Martín García--located in the Upper Río de la Plata, to the south of the Uruguay River--is an outcropping of the crystalline basement. Fourteen sampling sites were selected, five along the littoral section of the island and nine in inland ponds. Four major environmental variables were measured: water temperature, dissolved oxygen, electrical conductivity, and pH. A total of 34 species of Oligochaeta and Aphanoneura were found, 30 belonging to Naididae plus one species each of the Narapidae, Lumbricidae, Enchytraeidae, and Aeolosomatidae. The thirteen most frequent species were: A. leydigi (30%), N. bonettoi (13%), L. hoffmeisteri (11%), N. variabilis (10%), S. trivandrana (6.5%), A. pigueti (5.6%), D. sawayai (4.5%), D. digitata (3.5%), C. diastrophus (2.7%), A. costatus (2.5%), P. longiseta (2.0%), Enchytraeidae (1.5%), and A. p. paraguayensis (1.4%). UPGMA clustering of species based on their occurrence in different ecological conditions revealed two main species groups. Canonic-correspondence analysis (CCA) was conducted with the 15 most frequent and abundant species in the 9 sampling sites and the 4 environmental variables. Results from the CCA revealed that the order of fluctuation of the environmental variables during the sampling period was, from the greatest to the least: dissolved oxygen, conductivity, pH, and water temperature. Approximately 97.6% of the correlations between species and environmental variables were expressed on axis 1 of the ordination diagram. Species richness correlated with the four environmental variables in the following order, from the weakest to the strongest: water temperature, pH, electrical conductivity, and dissolved oxygen.

Communities of organisms inhabiting extreme terrestrial environments provide a unique opportunity to study evolutionary forces that drive population structure and genetic diversity under the combined challenges posed by multiple geogenic stressors. High abundance of an invasive pantropical earthworm (and the absence of indigenous lumbricid species) in the Furnas geothermal field (Sao Miguel Island, Azores) indicates its remarkable tolerance to high soil temperature, exceptionally high carbon dioxide and low oxygen levels, and elevated metal bioavailability, conditions which are lethal for the majority of terrestrial metazoans. Mitochondrial and nuclear markers were used to analyze the relationship between populations living inside and outside the geothermal field. Results showed that Pontoscolex corethrurus (Annelida, Oligochaeta, Glossoscolecidae) to be a genetically heterogeneous complex within the Sao Miguel landscape and is probably differentiated into cryptic species. The population exposed to the hostile soil conditions within the volcanic caldera possesses the lowest within-population mitochondrial diversity but an unexpectedly high degree of nuclear variability with several loci evidencing positive selection, parameters indicative of a genetically unique population only distantly related to conspecifics living outside the caldera. In conclusion, P. corethrurus inhabiting active volcanic soil is a discrete extremophile population that has evolved by tolerating a mixture of non-anthropogenic chemical and physical stressors.

Three new species from the Mexican states of Tabasco and Chiapas are added to the acanthodrilid earthworm genus Ramiellona, R. microscolecina sp. nov., R. tojolabala sp. nov. and R. teapaensis sp. nov. They belong to a group of species with penial setae and last pair of hearts in segment 12. All are holandric and the spermathecae have either a flat circular diverticle in a segment anterior to that of the ampulla (R. microscolecina sp. nov. and R. tojolabala sp. nov.) or two ovoidal and sessile diverticles on opposite sides in the same segment of the ampulla (R. teapaensis sp. nov.). Ramiellona americana (Gates) is re-described from a single specimen from central Guatemala, and the diagnosis of Ramiellona lasiura (Graff) from El Salvador is emended after reinvestigating a paratype specimen from the Senckenberg Naturmuseum Frankfurt. On the basis of several individuals from different populations of the Mexican states of Chiapas and Tabasco, the morphological variation of Ramiellona strigosa setosa Righi is described and its relationship with the Guatemalan Ramiellona strigosa strigosa Gates and Ramiellona eiseni (Michaelsen) is discussed. Finally, the position of Ramiellona within Acanthodrilidae and its relation to genera of the doubtful Octochaetidae is discussed.

Exotic earthworm introductions can alter above- and belowground properties of temperate forests, but the net impacts on forest soil carbon (C) dynamics are poorly understood. We used a mesocosm experiment to examine the impacts of earthworm species belonging to three different ecological groups (Lumbricus terrestris [anecic], Aporrectodea trapezoides [endogeic], and Eisenia fetida [epigeic]) on C distributions and storage in reconstructed soil profiles from a sandy temperate forest soil by measuring CO2 and dissolved organic carbon (DOC) losses, litter C incorporation into soil, and soil C storage with monospecific and species combinations as treatments. Soil CO2 loss was 30% greater from the Endogeic x Epigeic treatment than from controls (no earthworms) over the first 45 days; CO2 losses from monospecific treatments did not differ from controls. DOC losses were three orders of magnitude lower than CO2 losses, and were similar across earthworm community treatments. Communities with the anecic species accelerated litter C mass loss by 31-39% with differential mass loss of litter types (Acer rubrum > Populus grandidentata > Fagus grandifolia > Quercus rubra > or = Pinus strobus) indicative of leaf litter preference. Burrow system volume, continuity, and size distribution differed across earthworm treatments but did not affect cumulative CO2 or DOC losses. However, burrow system structure controlled vertical C redistribution by mediating the contributions of leaf litter to A-horizon C and N pools, as indicated by strong correlations between (1) subsurface vertical burrows made by anecic species, and accelerated leaf litter mass losses (with the exception of P. strobus); and (2) dense burrow networks in the A-horizon and the C and N properties of these pools. Final soil C storage was slightly lower in earthworm treatments, indicating that increased leaf litter C inputs into soil were more than offset by losses as CO2 and DOC across earthworm community treatments.

Phytochelatins are small cysteine-rich non-ribosomal peptides that chelate soft metal and metalloid ions, such as cadmium and arsenic. They are widely produced by plants and microbes; phytochelatin synthase genes are also present in animal species from several different phyla, but there is still little known about whether these genes are functional in animals, and if so, whether they are metal-responsive. We analysed phytochelatin production by direct chemical analysis in Lumbricus rubellus earthworms exposed to arsenic for a 28 day period, and found that arsenic clearly induced phytochelatin production in a dose-dependent manner. It was necessary to measure the phytochelatin metabolite concentrations directly, as there was no upregulation of phytochelatin synthase gene expression after 28 days: phytochelatin synthesis appears not to be transcriptionally regulated in animals. A further untargetted metabolomic analysis also found changes in metabolites associated with the transsulfuration pathway, which channels sulfur flux from methionine for phytochelatin synthesis. There was no evidence of biological transformation of arsenic (e.g. into methylated species) as a result of laboratory arsenic exposure. Finally, we compared wild populations of earthworms sampled from the field, and found that both arsenic-contaminated and cadmium-contaminated mine site worms had elevated phytochelatin concentrations.

Rhinodrilus alatus is an endemic giant earthworm of the Brazilian Cerrado hotspot used as live bait for about 80 years. The goal of this study was to gather ecological data about this species, which will support the establishment of management strategies. The life history, distribution and abundance of R. alatus were investigated in Cerrado, pastures and Eucalyptus plantation areas following the harvesting activities of the local extractors of this species. We found that this earthworm is abundant in all of the sampled areas, showing its resilience to land-use conversion. The Capture Per Unit Effort was 4.4 ± 5 individuals per 100 metres of transect and 5.6 ± 3 individuals per hour. The earthworm's annual cycle is markedly seasonal, with an aestivation period throughout the driest and coldest season of the year. Significant differences in the length and diameter of the body and in the diameter and depth of the aestivation chambers were found between the juveniles and adults. The distribution range of the species was expanded from two to 17 counties. The life history, abundance, distribution and resilience of R. alatus to certain perturbations are key elements to be considered in conservation and management strategies for this species.

Survival of earthworms in the environment depends on their ability to recognize and eliminate potential pathogens. This work is aimed to compare the innate defense mechanisms of two closely related earthworm species, Eisenia andrei and Eisenia fetida, that inhabit substantially different ecological niches. While E. andrei lives in a compost and manure, E. fetida can be found in the litter layer in forests. Therefore, the influence of environment-specific microbiota on the immune response of both species was followed. Firstly, a reliable method to discern between E. andrei and E. fetida based on species-specific primers for cytochrome c oxidase I (COI) and stringent PCR conditions was developed. Secondly, to analyze the immunological profile in both earthworm species, the activity and expression of lysozyme, pattern recognition protein CCF, and antimicrobial proteins with hemolytic function, fetidin and lysenins, have been assessed. Whereas, CCF and lysozyme showed only slight differences in the expression and activity, fetidin/lysenins expression as well as the hemolytic activity was considerably higher in E. andrei as compared to E. fetida. The expression of fetidin/lysenins in E. fetida was not affected upon the challenge with compost microbiota, suggesting more substantial changes in the regulation of the gene expression. Genomic DNA analyses revealed significantly higher level of fetidin/lysenins (determined using universal primer pairs) in E. andrei compared to E. fetida. It can be hypothesized that E. andrei colonizing compost as a new habitat acquired an evolutionary selection advantage resulting in a higher expression of antimicrobial proteins.

Phosphorus is one of the key factors that regulate soil fertility. Its deficiencies in soil are largely replenished by chemical fertilizers. The present study was aimed to isolate efficient phosphate solubilizing fungal strains from Eisenia fetida vermicompost. Out of total 30 fungal strains the most efficient phosphate solubilizing one was Emericella (Aspergillus) nidulans V1 (MTCC 11044), identified by custom sequencing of beta-tubulin gene and BLAST analysis. This strain solubilized 13 to 36% phosphate from four different rock phosphates. After three days of incubation of isolated culture with black Mussorie phosphate rock, the highest percentage of phosphate solubilization was 35.5 +/- 1.01 with a pH drop of 4.2 +/- 0.09. Kinetics of solubilization and acid production showed a linear relationship until day five of incubation. Interestingly, from zero to tenth day of incubation, solubility of soil phosphate increased gradually from 4.31 +/- 1.57 to 13.65 +/- 1.82 (mg kg(-1)) recording a maximum of 21.23 +/- 0.54 on day 45 in respect of the V1 isolate. Further, enhanced phosphorus uptake by Phaseolus plants with significant pod yield due to soil inoculation of Emericella nidulans V1 (MTCC 11044), demonstrated its prospect as an effective biofertilizer for plant growth.

This study aimed to develop bioavailability models for predicting Cu toxicity to earthworms (Lumbricus rubellus, Aporrectodea longa, and Eisenia fetida) in a range of soils of varying properties. A multicomponent Freundlich model, complying with the basic assumption of the biotic ligands model, was used to relate Cu toxicity to the free Cu(2+) activity and possible protective cations in soil porewater. Median lethal concentrations (LC50s) of Cu based on the total Cu concentration varied in each species from soil to soil, reaching differences of approximately a factor 9 in L. rubellus, 49 in A. longa and 45 in E. fetida. The relative sensitivity of the earthworms to Cu in different soils followed the same order: L. rubellus > A. longa > E. fetida. Only pH not other cations (K(+), Ca(2+), Na(+), and Mg(2+)) were found to exert significant protective effects against Cu toxicity to earthworms. The Freundlich-type model in which the protective effects of pH were included, explained 84%, 94%, and 96% of variations in LC50s of Cu (expressed as free ion activity) for L. rubellus, A. longa, and E. fetida, respectively. Predicted LC50s never differed by a factor of more than 2 from the observed LC50s. External validation of the model showed a similar level of precision, even though toxicity data for other soil organisms and for different endpoints were used. The findings of the present study showed the possibility of extrapolating the developed toxicity models for one earthworm species to another species. Moreover, the Freundlich-type model in which the free Cu(2+) activity and pH in soil porewater are considered can even be used to predict toxicity for other soil invertebrates and plants.

Aquatic oligochaetes have long been appreciated for their value in assessing habitat quality because they are ubiquitous sediment-dwelling filter feeders. Many oligochaete taxa are also important in the transmission of fish diseases. Distinguishing resistant and susceptible taxa is important for managing fish disease, yet challenging in practice. Tubifex tubifex (Oligochaeta: Tubificidae) is the definitive host for the complex life-cycle parasite, Myxobolus cerebralis, the causative agent of salmonid whirling disease. We developed two hydrolysis probe-based qualitative real-time PCR (qPCR) multiplex assays that distinguish among tubificid taxa collected from the Madison River, Montana, USA. The first assay distinguishes T. tubifex from Rhyacodrilus spp.; while the second classifies T. tubifex identified by the first assay into two genetic lineages (I and III). Specificity and sensitivity were optimized for each assay; the two assays showed specificity of 94.3% and 98.6% for the target oligochaetes, respectively. DNA sequencing verified the results. The development of these assays allowed us to more fully describe tubificid community composition (the taxa and their abundance at a site) and estimate the relative abundances of host taxa. To relate tubificid relative abundance to fish disease risk, we determined M. cerebralis infection prevalence in samples identified as T. tubifex using similar molecular techniques. Given prior information (i.e., morphological identification of sexually mature worms), Bayesian analysis inferred that the first qPCR assay improved taxonomic identification. Bayesian inference of the relative abundance of T. tubifex, combined with infection assay results, identified sites with a high prevalence of infected T. tubifex. To our knowledge, this study represents both the first assessment of oligochaete community composition using a qPCR assay based on fluorescent probes and the first use of Bayesian analysis to fully characterize the dominant infected taxa in streams where whirling disease is observed., (Copyright © 2013 Australian Society for Parasitology Inc. All rights reserved.)

The anoxic earthworm gut augments the activity of ingested microorganisms capable of anaerobiosis. Small earthworms (Lumbricidae) emit denitrification-derived N(2)O, whereas the large Octochaetus multiporus (Megascolecidae) does not. To examine this paradox, differently sized species of the families Glossoscolecidae (Rhinodrilus, Glossoscolex, Pontoscolex), Megascolecidae (Amynthas, Perionyx), Acanthodrilidae (Dichogaster), and Eudrilidae (Eudrilus) from Brazil were analyzed. Small species and the large Rhinodrilus alatus emitted N(2)O, whereas the large Glossoscolex paulistus did not, even though its gut could denitrify. N(2) and N(2)O were emitted concomitantly, and R. alatus emitted the highest amount of N(2). Denitrifiers and dissimilatory nitrate reducers were analyzed by barcoded amplicon pyrosequencing of narG, nirK, and nosZ. Gene sequences in gut and soil of the large G. paulistus were similar, whereas sequences in gut and soil of the small Amynthas gracilis were different and were also different compared with those of the gut and soil of G. paulistus. However, the denitrifying gut microbiota for both earthworms appeared to be soil-derived and dominated by Rhizobiales. The results demonstrated that (1) the emission of denitrification-derived N(2)O is widespread in different earthworm families, (2) large earthworms can also emit nitrogenous gases, and (3) ingested members of Rhizobiales are associated with this emission., (© 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)

Four new species of terrestrial earthworms belonging to the genus Amynthas were collected on the islands of Kinmen and Lieyu, Taiwan from March to November, 2008. They are Amynthas kinmenensis sp. nov., Amynthas wuhumontis sp. nov., Amynthas wujhouensis sp. nov., and Amynthas taiwumontis sp. nov. Amynthas kinmenensis sp. nov. is quadrithecal and is the most abundant earthworm widely distributed on the main island of Kinmen. It has numerous small genital papillae and is closely related to Amynthas polyglandularis (Tsai, 1964) from northern Taiwan. Amynthas wuhumontis sp. nov. is sexthecal and is distributed only in areas around Mt. Wuhu and Mt. Taiwu in east Kinmen. It has male pores each surrounded by three genital papillae: one anterior, one posterior and one medial. Amynthas wujhouensis sp. nov. and Amynthas taiwumontis sp. nov. are octothecal. The former has a sporadic distribution in Kinmen while the latter was only found in areas around Mt. Taiwu. Amynthas wujhouensis sp. nov. has a pair of large genital papillae closely adjacent to the crescent or semicircular shaped male porophores in XVIII. Amynthas taiwumontis sp. nov. has simple male pore structure and no genital papillae or genital markings. DNA barcodes (the 5' end sequences of the mitochondrial cytochrome c oxidase subunit 1 gene) from type specimens and other materials of the first three species are also reported.

Lake Hazar is an alkaline oligotrophic lake of tectonic origin, located in the Eastern Anatolia region in Turkey, 1248 a a.s.l. Its surface area is 80 km2, the average depth 93 m and maximum depth 205 m. The lake and its surroundings an under protection as a region of historical value. During the present study (2007-2012), samples were taken from 15 stations located at a depth of 2-200 m. Oligochaeta comprised 69% of the total invertebrate abundance. The profundal olgochaete fauna was found to consist of only three tubificid taxa, all of the subfamily Tubificinae. Potamothrix alatus hazaricus Timm & Arslan, n. ssp. was dominating anywhere down to maximum depths while Psammoryctides barbatus (Grube) and Ilyodrilus(?) sp. occurred seldom. All three are new records for Lake Hazar. Potamothrix alatus hazaricus shares the "winged" body shape in its genital region with the nominal, brackish-water subspecies P. a. alatus Finogenova, 1972, and the lateral position of the spermathecal pores and the shape of the ventral chaetae with the freshwater subspecies P. a. paravanicus Poddubnaja & Pataridze, 1989 known from Transcaucasian lakes. The mitochondrial COI barcoding gene suggests long separation between the two taxa, but the nuclear ITS region shows no variation. The generic position of Ilyodrilus (?) sp. remains obscure since its internal genitalia could not be studied.

Three new species and one new subspecies of earthworms from Hainan Island, China, are described: Amynthas dinganensis sp. nov., Amynthas tenuis sp. nov., Metaphire wuzhimontis sp. nov. and Metaphire magna minuscula subsp. nov. Amynthas dinganensis and Amynthas tenuis have two pairs of spermathecal pores in 5/6-6/7, and Metaphire wuzhimontis and Metaphire magna minuscula have two pairs of spermathecal pores in 7/8-8/9. COI and 16S gene fragments of the new species taxa have been sequenced. We provide comparisons between these new species taxa and related species using morphological and molecular characters.

A new clitellate species of Pristininae (Naididae), Pristina armata n. sp., found in the pitcher of the carnivorous plant Nepenthes sp., is reported from East Kalimantan, Indonesia. P. armata n. sp. is a very small clitellate, less than 1 mm long in fixed state, and without proboscis on the prostomium. Signs of reproduction by paratomy were observed, but the generic placement remains preliminary because sexually mature individuals were not found. P. armata n. sp. is characterized by giant hook-like dorsal chaetae at IV. The description of P. armata n. sp. was based on six fixed specimens of different size and stage of development. Noteworthy is the habitat of P. armata n. sp. in Nepenthes pitchers, this being the first clitellate species described from such a habitat. P. armata n. sp. may be a member of the nepenthebionts' community, realizing its life cycle inside the digestive fluid of the Nepenthes pitcher, or it belongs to nepenthephiles, species that commonly occur in this habitat but do not specialize on it.

Earthworm specimens collected from Mt. Chiak, National Park, Korea were found to represent three new species of megascolecid earthworms: Amynthas chiakensis sp. nov., Amynthas gyeongriae sp. nov., and Amynthas wonjuensis sp. nov. Amynthas chiakensis sp. nov. has two pairs of spermathecae in VII and VIII, small transverse oval male porophores each with C-shaped small patches of genital papillae, 0.22-0.31 circumference apart. Amynthas gyeongriae sp. nov. has spermathecae in VI-VIII, male field with large circular-shaped raised pads and seminal grooves, with pores 0.16-0.28 circumference apart. Amynthas wonjuensis sp. nov. has spermathecae in VI and VII, male pores superficial in XVIII on small oval-shaped white porophores lateral to large roughly circular raised genital papillae, 0.15 circumference apart.

Earthworms were collected in forests, damp habitats, springs, orchards and agricultural fields of the Kohgiluyeh & Boyer Ahmad Province, Iran, from April 2009 to April 2010. Specimens were collected at 20 established stations by digging and by diluted formalin methods. Ten species belonging to family Lumbricidae were identified based on morphology: Aporrectodea caliginosa (Savigny, 1826), Ap. rosea (Savigny, 1826), Ap. jassyensis (Michaelsen, 1891), Dendrobaena veneta (Rosa, 1886), D. byblica (Rosa, 1893) complex, D. orientalis orientalis Cernosvitov 1940, Eisenia fetida (Savigny, 1826), Eiseniella tetraedra (Savigny, 1826), Octolasion lacteum (Örley, 1881), Perelia kaznakovi (Michaelsen, 1910). Ap. caliginosa was the dominant species in this province and D. orientalis orientalis is a new record for Iran. A checklist of all earthworms species form Iran is presented, containing 19 species. Then, in order to show earthworm geographical affinities, hierarchical analysis were applied to available data on earthworm of Iran.

Five new species of terrestrial Enchytraeidae (Oligochaeta, Clitellata) are described from an experimental field area in Portugal. Achaeta coimbrensis sp. nov. belongs to a group of species without pyriform glands and with lateral spermathecal ectal pores. Fridericia sousai sp. nov., F. roembkei sp. nov., F. marginata sp. nov., and F. ciliotheca sp. nov. have a maximum of four chaetae per bundle and two spermathecal diverticula, a character combination shared by c. 30 other species of this genus. The new Fridericia species are distinguished from these congeners by combinations of characters, but the ventral pattern of the clitellum alone is sufficient to separate the new species from each other. Enchylea heteroducta Nielsen & Christensen, 1963 is redescribed, this being the first record after the original description and the first record from a natural habitat. Further 16 species of enchytraeids are recorded, and there are now 32 species of enchytraeids known from Portugal.

We designed a field survey (the entire river length; not dammed: First and Second plateaus; dammed: Third Plateau) to test the hypothesis that the cascade of reservoirs promoted a reduction in species richness and changed the composition of Oligochaeta assemblage along the Iguaçu River. Changes in environmental variables and in richness and composition of Oligochaeta were summarized by Canonic Correspondence Analysis. Along the Iguaçu River, conductivity, and altitude decreased, whereas temperature increased. Oligochaeta composition showed a significant spatial variation, with higher abundances of the family Tubificidae and the genus Dero (Naididae) occurring in the First Plateau. In the Second and Third plateaus, few species were dominant, with increases in the presence of species of Naididae below dams. We found a clear decrease in species richness along the Iguaçu River. Moreover, we found that Oligochaeta assemblage was influenced by some environmental variables such as altitude, conductivity, substrate type and temperature, and by anthropogenic activities (human occupation and damming). The results supported the use of Oligochaeta as surrogate taxa to predict environmental changes along impacted (dammed and eutrophic) rivers. The validity of this was indicated by the strong and significant gradient registered, from the headwater to mouth of the Iguaçu River.

In tropical savannas, the earthworm communities have a predominant role since they regulate the soil structure and dynamics of the organic matter. To study the effect on earthworm populations in two differently managed savannas, we compared the general aspects of the biology and ecology of earthworm populations from a 40 years protected savanna (SP) with no fire or cattle raising at the Estaci6n Biol6gica de los Llanos, Venezuela (EBLL), and a natural savanna (SNI), under normal burning and cattle raising management conditions. Sampling was carried out at the end of the dry season (April), and at the peak of the wet season (July-August). The main physical properties of soils per system were estimated. In each system, in plots of 90x90m, five fixed sampling units were selected at random; and at each sampling point one soil monolith of 25x25x30cm was collected per unit. Earthworms were extracted using the hand sorting extraction method; and the flotation method was used to estimate the density of cocoons. The earthworms were classified in different ecological categories considering their pigmentation, size and depth profile distribution. As a result of the savanna protection, physical parameters were modified in relation to SNI. The SP soils had higher soil moisture when compared to SNI. Soil moisture varied with depth during the dry season since, after the start of the rainy season, the soil was saturated. Field capacity in the SP was greater than that in the SNI. The surface apparent bulk density of soil was lower in the SP respect SNI, reflecting a lower soil compaction. Total average for the density and biomass of earthworms differed greatly, showing higher values in the SP. The earthworm density average in SP ranged between 25.6-85 individuals/m2 and the average biomass between 6.92-23.23g/m2. While in SNI, earthworms were only found in August, with a mean density of 22.40individuals/m2 and a mean biomass of 5.17g/m2. The vertical distribution pattern was only analyzed for the SP, and as in the SNI, no earthworms were found during the dry season; in the SP, a migration of earthworms to deeper layers during the dry season was observed; while in the rainy season they moved to upper soil layers to feed. The systems were characterized by abundance in juvenile Glossoscolecidae family earthworms which might be included in the endo-anecic ecological category. Results suggest that savanna agricultural management can modify an important fraction of pedofauna, particularly their earthworm communities, by modification of soil properties.

The importance of the Aporrectodea caliginosa species complex lies in the great abundance and wide distribution of the species which exist within it. For more than a century, chaos has surrounded this complex; morphological criteria has failed to solve the taxonomic status of these species. This present body of work aims to study the phylogeny of this complex by increasing the number of samples used in previous molecular works and by including morphologically-similar species that were never studied using molecular tools (A. giardi, Nicodrilus monticola, N. carochensis and N. tetramammalis). Two basal clades were obtained: one formed by A. caliginosa and A. tuberculata and the other by the rest of the species. This second clade was divided into two more: one with Eurosiberian and another with Mediterranean forms. A. caliginosa and A. longa were divided into two paraphyletic groups. Both A. giardi and A. nocturna showed characteristics consistent with monophyletic groups. Each of the two recovered lineages of A. trapezoides were phylogenetically related to different sexual species. While lineage I of A. trapezoides was monophyletic, lineage II resulted to be paraphyletic, as well as the three Nicodrilus 'species'. The diversification of the complex occurred during the Late Miocene-Early Pliocene (6.92-11.09 Mya). The parthenogenetic forms within the Mediterranean clade would have diversified before the ones in the Eurosiberian clade (3.13-4.64 Mya and 1.05-3.48 Mya, respectively), thus implying the existence not only of at least two different moments in which parthenogenesis arose within this complex of species, but also of two different and independent evolutionary lines. Neither the 4× rule nor the GMYC method for species delimitation were successful for distinguishing taxonomically-distinct species., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Many recent studies on invertebrates have shown how morphology not always captures the true diversity of taxa, with cryptic speciation often being discussed in this context. Here, we show how diversification patterns can be very different in two clades of closely related earthworms in the genus Hormogaster stressing the risk of using nonspecific substitution rate values across taxa. On the one hand, the Hormogaster elisae species complex, endemic to the central Iberian Peninsula, shows morphological stasis. On the other hand, a clade of Hormogaster from the NE Iberian Peninsula shows an enormous morphological variability, with 15 described morphospecies. The H. elisae complex, however, evolves faster genetically, and this could be explained by the harsher environmental conditions to which it is confined-as detected in this study, that is, sandier and slightly poorer soils with lower pH values than those of the other species in the family. These extreme conditions could be at the same time limiting morphological evolution and thus be responsible for the observed morphological stasis in this clade. Contrarily, Hormogaster species from the NE Iberian Peninsula, although still inhabiting harsher milieu than other earthworm groups, have had the opportunity to evolve into a greater morphological disparity. An attempt to delimit species within this group following the recently proposed general mixed Yule-coalescent method showed a higher number of entities than expected under the morphospecies concept, most probably due to the low vagility of these animals, which considerably limits gene flow between distant conspecific populations, but also because of the decoupling between morphological and genetic evolution in the H. elisae complex., (© 2012 Blackwell Publishing Ltd.)

An incubation experiment was conducted to study the changes that occur in organic carbon content, phosphorous and potassium availability and other soil properties with ingestion of soil mixed with rubber leaf litter and cow dung by five earthworm species viz. Pontoscolex corethrurus, Drawida assamensis, Drawida papillifer papillifer, Eutyphoeus comillahnus and Metaphire houlletiof rubber plantation in Tripura (India). Due to earthworm activity organic C (1.56-1.63%) and available P (14.71-27.60 mg 100 g(-1)) and K (43.50-49.0 mg 100 g(-1)) content of the soil increased significantly (p < 0.05) in most of the earthworm species studied. M. houlleti and D. papillifer papillifer had the highest P (27.60 mg 100 g ) and K (49.0 mg 100 g ) mobilization capacity, respectively. Earthworms, irrespective of the species, increased the pH (7.05-7.17) and electrical conductivity (663-1383 microS cm(-1)) of the soil significantly (p < 0.05).

Earthworms are known for their important role within the functioning of an ecosystem, and their diversity can be used as an indicator of ecosystem health. To date, earthworm diversity has been investigated through conventional extraction methods such as handsorting, soil washing or the application of a mustard solution. Such techniques are time consuming and often difficult to apply. We showed that combining DNA metabarcoding and next-generation sequencing facilitates the identification of earthworm species from soil samples. The first step of our experiments was to create a reference database of mitochondrial DNA (mtDNA) 16S gene for 14 earthworm species found in the French Alps. Using this database, we designed two new primer pairs targeting very short and informative DNA sequences (about 30 and 70 bp) that allow unambiguous species identification. Finally, we analysed extracellular DNA taken from soil samples in two localities (two plots per locality and eight samples per plot). The two short metabarcode regions led to the identification of a total of eight earthworm species. The earthworm communities identified by the DNA-based approach appeared to be well differentiated between the two localities and are consistent with results derived from inventories collected using the handsorting method. The possibility of assessing earthworm communities from hundreds or even thousands of localities through the use of extracellular soil DNA will undoubtedly stimulate further ecological research on these organisms. Using the same DNA extracts, our study also illustrates the potential of environmental DNA as a tool to assess the diversity of other soil-dwelling animal taxa., (© 2012 Blackwell Publishing Ltd.)

This study provides detailed information on the invertebrate hosts of Myxobolus pseudodispar (Myxozoa) and explores the susceptibility range of several species and analyses the relevance of the species composition of an oligochaete population. Our findings demonstrate that the oligochaete host range of M. pseudodispar is similarly wide as the number of vertebrate host species. Besides Tubifex tubifex and Limnodrilus hoffmeisteri, Psammoryctides barbatus and Psammoryctides moravicus were also found to be susceptible invertebrate hosts. The genetic characterization of the mitochondrial 16S rDNA of T. tubifex sensu lato revealed that lineages I, II and III are susceptible to M. pseudodispar, whereas T. tubifex lineage VI seems to be non-susceptible. T. tubifex lineage V and L. hoffmeisteri specimens were positive in a M. pseudodispar-specific PCR, but in most cases, the release of mature actinospores could not be detected. Hence, these non-susceptible oligochaetes likely serve as `biological filters` as they remove myxospores from the sediment without producing actinospores. Together with the phylogenetic analysis of the susceptible and non-susceptible oligochaete hosts on the basis of mt 16S rDNA sequences, the route of the development of M. pseudodispar in the oligochaete hosts was tracked by in situ hybridization. According to our findings, the gut epithelia seem to be a portal of entry of the sporoplasms, where the development of the parasite also takes place. The basal lamina seems to be involved in the migration of the parasite, and the worm's cellular immune response is activated by the infection., (© 2012 Blackwell Publishing Ltd.)

Little quantitative ecological information exists on the diets of most invertebrate feeding reptiles, particularly nocturnal or elusive species that are difficult to observe. In the UK and elsewhere, reptiles are legally required to be relocated before land development can proceed, but without knowledge of their dietary requirements, the suitability of receptor sites cannot be known. Here, we tested the ability of non-invasive DNA-based molecular diagnostics (454 pyrosequencing) to analyse reptile diets, with the specific aims of determining which earthworm species are exploited by slow worms (the legless lizard Anguis fragilis) and whether they feed on the deeper-living earthworm species that only come to the surface at night. Slow worm faecal samples from four different habitats were analysed using earthworm-specific PCR primers. We found that 86% of slow worms (N=80) had eaten earthworms. In lowland heath and marshy/acid grassland, Lumbricus rubellus, a surface-dwelling epigeic species, dominated slow worm diet. In two other habitats, riverside pasture and calciferous coarse grassland, diet was dominated by deeper-living anecic and endogeic species. We conclude that all species of earthworm are exploited by these reptiles and lack of specialization allows slow worms to thrive in a wide variety of habitats. Pyrosequencing of prey DNA in faeces showed promise as a practical, rapid and relatively inexpensive means of obtaining detailed and valuable ecological information on the diets of reptiles., (© 2011 Blackwell Publishing Ltd.)

DNA barcoding remains a challenge when applied to diet analyses, ancient DNA studies, environmental DNA samples and, more generally, in any cases where DNA samples have not been adequately preserved. Because the size of the commonly used barcoding marker (COI) is over 600 base pairs (bp), amplification fails when the DNA molecule is degraded into smaller fragments. However, relevant information for specimen identification may not be evenly distributed along the barcoding region, and a shorter target can be sufficient for identification purposes. This study proposes a new, widely applicable, method to compare the performance of all potential 'mini-barcodes' for a given molecular marker and to objectively select the shortest and most informative one. Our method is based on a sliding window analysis implemented in the new R package SPIDER (Species IDentity and Evolution in R). This method is applicable to any taxon and any molecular marker. Here, it was tested on earthworm DNA that had been degraded through digestion by carnivorous landsnails. A 100 bp region of 16 S rDNA was selected as the shortest informative fragment (mini-barcode) required for accurate specimen identification. Corresponding primers were designed and used to amplify degraded earthworm (prey) DNA from 46 landsnail (predator) faeces using 454-pyrosequencing. This led to the detection of 18 earthworm species in the diet of the snail. We encourage molecular ecologists to use this method to objectively select the most informative region of the gene they aim to amplify from degraded DNA. The method and tools provided here, can be particularly useful (1) when dealing with degraded DNA for which only small fragments can be amplified, (2) for cases where no consensus has yet been reached on the appropriate barcode gene, or (3) to allow direct analysis of short reads derived from massively parallel sequencing without the need for bioinformatic consolidation.

Traditional earthworm taxonomy is hindered due to their anatomical simplicity and the plasticity of the characteristics often used for diagnosing species. Making phylogenetic inferences based on these characters is more than difficult. In this study we use molecular tools to unravel the phylogeny of the clitellate family Hormogastridae. The family includes species of large to mid-sized earthworms distributed almost exclusively in the western Mediterranean region where they play an important ecological role. We analyzed individuals from 46 locations spanning the Iberian Peninsula to Corsica and Sardinia, representing the four described genera in the family and 20 species. Molecular markers include mitochondrial regions of the cytochrome c oxidase subunit I gene (COI), 16S rRNA and tRNAs for Leu, Ala, and Ser, two nuclear ribosomal genes (nearly complete 18S rRNA and a fragment of 28S rRNA) and two nuclear protein-encoding genes (histones H3 and H4). Analyses of the data using different approaches corroborates monophyly of Hormogastridae, but the genus Hormogaster is paraphyletic and Hormogaster pretiosa appears polyphyletic, stressing the need for taxonomic revisionary work in the family. The genus Vignysa could represent an early offshoot in the family, although the relationships with other genera are uncertain. The genus Hemigastrodrilus is related to the Hormogaster elisae complex and both are found in the Atlantic drainage of the Iberian Peninsula and France. From a biogeographic perspective Corsica and Sardinia include members of two separate hormogastrid lineages. The species located in Corsica and Northern Sardinia are related to Vignysa, whereas Hormogaster pretiosa pretiosa, from Southern Sardinia, is closely related to the Hormogaster species from the NE Iberian Peninsula. A molecular dating of the tree using the separation of the Sardinian microplate as a calibration point (at 33 MY) and assuming a model of vicariance indicates that the diversification of Hormogastridae may be ancient, ranging from 97 to 67 Ma., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Studies on community structures of earthworms of rubber (Hevea brasiliensis) plantations and their adjacent mixed forests in West Tripura (India) revealed that both the studied sites harvoured 10 earthworm species. Nine species (Pontoscolex corethrurus, Kanchuria sp 1, Metaphire houlleti, Drawida papillifer papillifer, Drawida assamensis, Gordiodrilus elegans, Eutyphoeus assamensis, Eutyphoeus comillahnus and Eutyphoeus gigas) were common to both. While Octochaetona beatrixwas found only in the rubber plantations, Dichogaster affinis was restricted to the mixed forest only. Earthworms were found mostly within 15 cm depth of soils having mean temperature of 27 degrees C, moisture of 23%, pH of 4.57, organic matter of 1.34% and water holding capacity of 36%. Mean earthworm density in rubber plantations (115 ind. m(-2)) was significantly higher (p = 0.003, t = 3.83) than that in the mixed forests (69 ind. m(-2)) due to dominance of Pontoscolex corethrurus, an exotic species. Numbers of dominant species were two (P. corethrurus and D. assamensis) in the rubber plantations and five (P. corethrurus D. assamensis, D. papilliferpapillifer, M. houlleti and Kanchuria sp 1) in the mixed forests. Compared to the mixed forests, significantly low (p<0.05) Shannon diversity index (H) and species evenness and high index of dominance in the rubber plantation were evaluated.

The population of soil mesofauna in the basin of the small river subzone of the northern taiga (Karelia) has been investigated. It was shown that indexes of the number and mass of soil mesofauna in the landscape-ecological row ofbiogeocenosises are maximal in floodplain soils. The taxonomic composition and structure of domination of the soil mesofauna population depends on the location of biogeocenosis in the landscape: earthworms are dominants in riverine floodplain biogeocenosises, and larvae of elaterids and spiders prevail in the places outside of floodplains. The abundance of saprophytic invertebrates in floodplain biogeocenosises results in formation of humus of the mull type. A group of animals with mixed type of nutrition dominates in the places outside of floodplain soils that are related with humus of the moder-mor type. The population of rove beetles (Staphylinidae) allows the division of biogeocenosises into two groups according to their position in the landscape.

To investigate whether coelomic fluid secreted by earthworms can be a noninvasive source of DNA, we amplified and sequenced DNA extracted from the coelomic fluid and muscle tissue of eight worms. The sequences obtained using DNA extracted from both sources were identical. All cytochrome c oxidase subunit I (COI) mitochondrial DNA sequences, including those retrieved from GenBank, formed a monophyletic group of Metaphire sieboldi. The results indicate that we successfully extracted total DNA from coelomic fluid secreted by earthworm., (© 2011 Blackwell Publishing Ltd.)

The Oligochaeta forms an important part of the macroinvertebrates inhabiting sediments of lotic ecosystems. It has an important role in the cycling of matter and energy transfer in these environments. The aim of this study is to analyse limnological variables, their influence on the structure and diversity of benthic oligochaete taxocenosis in a stretch of the Upper São Francisco River and its tributary the Piumhi River. Samples were taken in two climatic periods, the dry season in October 2006 and 2007 and the rainy season in March 2007 and 2008 at three points along the Piumhi River and six points along the São Francisco River. The sediment of the São Francisco consisted predominantly of sand and clay, whereas the sediment of the Piumhi was mainly sandy. Six species of oligochaete occurred in the Piumhi River while seven were found in the São Francisco. Of these, Pristina synclites Stephenson, 1925, Pristina americana Cernosvitov, 1937, Bothrioneurum sp. Stolc, 1888 and Limnodrilus hoffmeisteri Claparede, 1862 occurred in the sediment of both rivers. L. hoffmeisteri showed the highest numerical abundance in the Piumhi River and Brinkhurstia americana (Brinkhurst, 1964) and L. neotropicus Cernosvitov, 1939 were the most abundant species in the São Francisco River. The highest oligochaete density was recorded in the Piumhi during the dry seasons. Canonical correspondence analyses (CCA) of sediment characteristics explained most of the data variability and the association of the presence of oligochaete species in the Piumhi and São Francisco Rivers with the limnological variables (grain size composition and total nitrogen and phosphorus concentrations in the sediment).

We have constructed the first ever phylogeny for the New Zealand earthworm fauna (Megascolecinae and Acanthodrilinae) including representatives from other major continental regions. Bayesian and maximum likelihood phylogenetic trees were constructed from 427 base pairs from the mitochondrial large subunit (16S) rRNA gene and 661 base pairs from the nuclear large subunit (28S) rRNA gene. Within the Acanthodrilinae we were able to identify a number of well-supported clades that were restricted to continental landmasses. Estimates of nodal support for these major clades were generally high, but relationships among clades were poorly resolved. The phylogenetic analyses revealed several independent lineages in New Zealand, some of which had a comparable phylogenetic depth to monophyletic groups sampled from Madagascar, Africa, North America and Australia. These results are consistent with at least some of these clades having inhabited New Zealand since rifting from Gondwana in the Late Cretaceous. Within the New Zealand Acanthodrilinae, major clades tended to be restricted to specific regions of New Zealand, with the central North Island and Cook Strait representing major biogeographic boundaries. Our field surveys of New Zealand and subsequent identification has also revealed extensive cryptic taxonomic diversity with approximately 48 new species sampled in addition to the 199 species recognized by previous authors. Our results indicate that further survey and taxonomic work is required to establish a foundation for future biogeographic and ecological research on this vitally important component of the New Zealand biota., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Background: As currently defined, the genus Postandrilus Qui and Bouché, 1998, (Lumbricidae) includes six earthworm species, five occurring in Majorca (Baleares Islands, western Mediterranean) and another in Galicia (NW Spain). This disjunct and restricted distribution raises some interesting phylogeographic questions: (1) Is Postandrilus distribution the result of the separation of the Baleares-Kabylies (BK) microplate from the proto-Iberian Peninsula in the Late Oligocene (30-28 Mya)--vicariant hypothesis? (2) Did Postandrilus diversify in Spain and then colonize the Baleares during the Messinian salinity crisis (MSC) 5.96-5.33 Mya--dispersal hypothesis? (3) Is the distribution the result of a two-step process--vicariance with subsequent dispersal?, Methodology/principal Findings: To answer these questions and assess Postandrilus evolutionary relationships and systematics, we collected all of the six Postandrilus species (46 specimens - 16 locations) and used Aporrectodea morenoe and three Prosellodrilus and two Cataladrilus species as the outgroup. Regions of the nuclear 28S rDNA and mitochondrial 16S rDNA, 12S rDNA, ND1, COII and tRNA genes (4,666 bp) were sequenced and analyzed using maximum likelihood and Bayesian methods of phylogenetic and divergence time estimation. The resulting trees revealed six new Postandrilus species in Majorca that clustered with the other five species already described. This Majorcan clade was sister to an Iberian clade including A. morenoe (outgroup) and Postandrilus bertae. Our phylogeny and divergence time estimates indicated that the split between the Iberian and Majorcan Postandrilus clades took place 30.1 Mya, in concordance with the break of the BK microplate from the proto-Iberian Peninsula, and that the present Majorcan clade diversified 5.7 Mya, during the MSC., Conclusions: Postandrilus is highly diverse including multiple cryptic species in Majorca. The genus is not monophyletic and invalid as currently defined. Postandrilus is of vicariant origin and its radiation began in the Late Oligocene.

Majority of municipal (urban) solid waste (MSW) is disposed of in landfills (anaerobic composting). However, this disposal system is reported to produce hazardous environmental impacts and new policies are initiated to protect the environment from such impacts by discouraging the practice of disposal of solid waste in landfills. Eco-friendly disposal alternatives to landfills need to be explored. One of the technological options for treatment and disposal of organic solid wastes is vermicomposting. Commercial vermicomposting is reported to be practicable for treatment and disposal of many organic solids and byproducts in agricultural production and processing industries. However, this alternative has not been tried for MSW on large scale. This paper highlights the application of vermicomposting for treatment of organic solid waste, generated at urban residential area at Pune [organic component of this urban solid waste (MSW)]. Vermicomposting was tried on this segregated solid waste using exotic species of earthworm--Eudrilus eugeniae--commonly called 'African Night Crawler'. Bench scale reactor studies were carried out on organic solid waste under controlled optimum environmental conditions (moisture content: 48-52 percent, pH: 7.0-7.2, temperature: ambient), with variable vermi-loading [40-80 g of worms/kg of urban solid waste (MSW)]. Characteristics of solid waste were monitored through conventional parameters and additional environmental parameters like BOD5 and COD. The results of investigative studies are encouraging and indicate that organic solid waste can be treated in a reasonable period of 32-34 days through vermicomposting with around 60 percent reduction in the volume.

This work summarizes the observations on 30 species of microdriles belonging to the families Naididae (Rhyacodrilinae, Pristininae, Naidinae, Phallodrilinae, and Tubificinae), Phreodrilidae, Lumbriculidae, and Enchytraeidae using scanning electron microscopy. The lumbricid Eiseniella tetraedra, a megadrile species common in typical microdrile habitats, was used for comparison. Microdriles display external ciliate sense structures along the entire body; even at the clitellum and in budding and regeneration zones. According to the shape of the cilia, these sense structures can be divided into receptors of blunt cilia, receptors of sharp cilia, and composed receptors. Sense receptors can be morphologically unconspicuous or clearly defined on sensory buds or papillae. All microdriles studied have receptors of blunt cilia. Enchytraeids have characteristic receptors of short cilia. Pristina (Pristininae), Chaetogaster, Ophidonais, and Stylaria (Naidinae) have receptors of long blunt cilia. Composed receptors were found only in some microdriles and E. tetraedra. Receptors of sharp cilia have been found in most microdriles. Enchytraeids might be the only exception, but sharp cilia are probably present in the amphibiotic Cognettia sphagnetorum. Sensory cells with long sharp cilia might play a rheoreceptor role, and their presence in E. tetraedra and C. sphagnetorum would imply the reappearing of an ancient character that was probably lost with the transit from aquatic to terrestrial habitats. Some lumbriculids have ciliated fields. Anatomically, these structures appear as intermediate between the typical isolate sensory structures of microdriles and the sensillae of the hirudineans. The general pattern in microdriles is that uniciliate receptors and multiciliate receptors are separated, which supports the presumed aquatic origin of the clitellates., (© 2010 Wiley-Liss, Inc.)

MicroRNAs are a recently identified class of small regulatory RNAs that target more than 30% protein-coding genes. Elevating evidence shows that miRNAs play a critical role in many biological processes, including developmental timing, tissue differentiation, and response to chemical exposure. In this study, we applied a computational approach to analyze expressed sequence tags, and identified 32 miRNAs belonging to 22 miRNA families, in three earthworm species Eisenia fetida, Eisenia andrei, and Lumbricus rubellus. These newly identified earthworm miRNAs possess a difference of 2-4 nucleotides from their homologous counterparts in Caenorhabditis elegans. They also share similar features with other known animal miRNAs, for instance, the nucleotide U being dominant in both mature and pre-miRNA sequences, particularly in the first position of mature miRNA sequences at the 5' end. The newly identified earthworm miRNAs putatively regulate mRNA genes that are involved in many important biological processes and pathways related to development, growth, locomotion, and reproduction as well as response to stresses, particularly oxidative stress. Future efforts will focus on experimental validation of their presence and target mRNA genes to further elucidate their biological functions in earthworms., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The effects on macro-benthos and benthos environment of planting 200 hm2 water hyacinth (E. crassipens) in Zhushan Bay, Lake Taihu, were studied during 8-10 months consecutive surveys. Results indicated that average densities of mollusca (the main species were Bellamya aeruginosa) in far-planting, near-planting and planting area were 276.67, 371.11 and 440.00 ind/m2, respectively, and biomass were 373.15, 486.57 and 672.54 g/m2, respectively, showed that average density and biomass of planting area's were higher than those of others. However, the average density and biomass of Oligochaeta (the main species was Limodrilus hoffmeisteri) and Chironomidae in planting area were lower than that of outside planting area. The density and biomass of three dominant species of benthic animal increased quickly during 8-9 months, decreased quickly in October inside and outside water hyacinth planting area. The reason of this phenomenon could be possible that lots of cyanobacteria cells died and consumed dissolve oxygen in proceed decomposing. Algae cells released lots of phosphorus and nitrogen simultaneously, so macro-benthos died in this environment. The indexes of Shannon-Weaver and Simpson indicated that water environment was in moderate polluted state. On the basis of the survey results, the large-area and high-density planting water hyacinth haven't demonstrated a great impact on macrobenthos and benthos environment in short planting time (about 6 months planting time).

The aim of this study was to evaluate the presence of Chaetogaster limnaei K. von Baer, 1927 in four species of mollusks collected in irrigation channels of a horticultural garden in the city of Juiz de Fora, Minas Gerais State, Southeastern Brazil. The collections were made from June 2005 to January 2006. Of the four species of mollusks analysed, we could not detect the presence of C. limnaei in Pomacea lineata (Spix, 1827). There were recorded 320 specimens of C. limnaei in association with Aplexa rivalis Mastou & Rackett, 1898, Lymnaea columella Say, 1818 and Biomphalaria sp., the latter having the highest number of associated Naididae.

A multigene data set (12S, 16S, and COI mitochondrial DNA; 18S and 28S nuclear DNA) was analyzed by Bayesian inference to estimate the phylogeny of a sample of the clitellate family Enchytraeidae (86 species representing 14 nominal genera). Monophyly, as well as a basal dichotomy, of the family Enchytraeidae obtained maximum support, with one clade containing Hemienchytraeus and Achaeta, the other the remaining 12 genera analysed. The latter group is basally resolved in several well-supported clades. Lumbricillus and Grania are closely related. Bryodrilus, Oconnorella, Henlea and two species of Marionina (M. cf. riparia, and M. communis) form a well-supported clade. Cognettia is sister to Stercutus, and Cernosvitoviella sister to Mesenchytraeus, and the four together appear to be a monophyletic group. A large part of the taxonomically problematic Marionina appears to be a group not closely related to the type species (M. georgiana), and this group also includes Enchytronia. Further, this Marionina/Enchytronia group appears to be sister to a clade comprising the more or less littoral marine genera Stephensoniella and Enchytraeus. Hemifridericia, Buchholzia and Fridericia, the three genera characterized by two types of coelomocytes, also form a well-supported clade. The study corroborates most of the multi-species genera analysed (Cognettia, Cernosvitoviella, Mesenchytraeus, Oconnorella, Henlea, Enchytraeus, Grania, Buchholzia and Fridericia); only Lumbricillus and Marionina are non-monophyletic as currently defined., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Limnodrilus simplex sp. nov. (Oligochaeta: Naididae: Tubificinae) is described based on a single specimen from the mainstream of the Changjiang River near Anqing City, Anhui Province, China. The new species is assigned to Limnodrilus by the presence of long vasa deferentia, spindle-shaped atria with long ejaculatory ducts, large prostate glands, and thick cylindrical penial sheaths. It differs from its congeners in having simple-pointed chaetae and cuticularized penial sheaths without hoods. Limnodrilus simplex is closer to L. paramblysetus and L. amblysetus in possessing penial sheaths with relatively low length/maximum width ratio.

On a local scale, invasiveness of introduced species and invasibility of habitats together determine invasion success. A key issue in invasion ecology has been how to quantify the contribution of species invasiveness and habitat invasibility separately. Conventional approaches, such as comparing the differences in traits and/or impacts of species between native and/or invaded ranges, do not determine the extent to which the performance of invaders is due to either the effects of species traits or habitat characteristics. Here we explore the interaction between two of the most widespread earthworm invaders in the world (Asian Amynthas agrestis and European Lumbricus rubellus) and study the effects of species invasiveness and habitat invasibility separately through an alternative approach of "third habitat" in Tennessee, USA. We propose that feeding behaviors of earthworms will be critical to invasion success because trophic ecology of invasive animals plays a key role in the invasion process. We found that (1) the biomass and isotopic abundances (delta13C and delta15N) of A. agrestis were not impacted by either direct effects of L. rubellus competition or indirect effects of L. rubellus-preconditioned habitat; (2) A. agrestis disrupted the relationship between L. rubellus and soil microorganisms and consequently hindered litter consumption by L. rubellus; and (3) compared to L. rubellus, A. agrestis shifted its diet more readily to consume more litter, more soil gram-positive (G+) bacteria (which may be important for litter digestion), and more non-microbial soil fauna when soil microorganisms were depleted. In conclusion, A. agrestis showed strong invasiveness through its dietary flexibility through diet shifting and superior feeding behavior and its indirectly negative effect of habitat invasibility on L. rubellus via changes in the soil microorganism community. In such context, our results expand on the resource fluctuation hypothesis and support the superior competitor hypothesis. This work presents additional approaches in invasion ecology, provides some new dimensions for further research, and contributes to a greater understanding of the importance of interactions between multiple invading species.

Species delimitation of earthworms has been difficult to determine with certainty due to their structural simplicity. We sequenced fragments of COI, 16S, t-RNAs and 28S for 202 Hormogastridae individuals from the central Iberian Peninsula and three outgroup taxa. A morphological constancy was found but a high genetic diversity suggests the presence of five cryptic allopatric species. Results showed a pattern of isolation by distance and a positive but weak correlation between some soil properties (coarse sand and total loam content) and genetic distances, which indicates that these populations may have been shaped genetically but not morphologically, by the environment., (Copyright 2010 Elsevier Inc. All rights reserved.)

A comparative study was conducted two epigeic species earthworms (Bimastus parvus and Eisenia foetida) for the evaluation of their efficacy in vermicomposting of sewage sludge. The various changes studied during pot experiments were the physiochemical properties of the sewage sludge, sludge reduction and earthworm biomass. Vermicomposting resulted that both epigeic species earthworms showed same capability among sewage sludge mineralization and decomposition rate and reduction. By the end of experiment, the pH value declined to 6.27 with B. parvus and 7.07 with E. foetida, but both epigeic species earthworms showed same mineralization and decomposition rate. B. parvus produced 31.96%, 5.76% and 17.91% increases in nitrogen, phosphorus, and potassium as well as 44.14% and 30.69% decreases in C/N and C/P ratios as compared to initial after 30 days of inoculation. In contrast, E. foetida produced 35.48% and 11.58% increases in nitrogen and potassium as well as 10.12%, 46.73% and 20.50% decreases in phosphorus, C/N and C/P ratios as compared to initial after 30 days of earthworm activity. At the same time, both epigeic species earthworms resulted in significant reduction in heavy metal content. The reduction in heavy metal content for B. parvus and E. foetida was found in the order: Zn > Cu > Pb > Cr and Cu > Zn > Ph > Cr. At the end of experiment, the weight and cocoons of B. parvus and E. foetida showed significant increase, which the growth rate and the reproductive rate were 76%-86% and 156%-131% respectively.

Unlabelled: Contact bioassays are important for testing the ecotoxicity of solid materials. However, survival and reproduction tests are often not practical due to their duration which may last for several weeks. Avoidance tests with soil invertebrates may offer an alternative or extension to the classic test batteries due to their short duration (days rather than weeks) and due to a sensitive sub-acute endpoint (behavior)., The Aims of Our Study Were: (a) to evaluate the effects of three solid industrial wastes (incineration ash, contaminated wood chips and contaminated soil) on three Oligochaeta species (enchytraeids Enchytraeusalbidus, Enchytraeus crypticus and earthworm Eisenia fetida) in avoidance tests; (b) to compare the sensitivity among the species and to compare results of avoidance test to reproduction tests; (c) to elucidate if measuring the weight in the earthworm avoidance test could be reasonable additional endpoint. Avoidance mostly increased with the increasing percent of waste in the mixture showing a dose-response curve. E. fetida was the most sensitive species and E. crypticus the least one. An additional endpoint, (changes in weight after two-day exposure) was not found to be more sensitive than avoidance reaction, but it confirmed that earthworms staying in the highest concentrations of the waste mixture were affected showing apparent weight reduction. Our results indicate that avoidance tests with earthworms and enchytraeids are feasible for waste testing., (Copyright 2009 Elsevier Ltd. All rights reserved.)

This paper reviews the role of earthworms in the early colonisation of contaminated soils as well as sediment and waste deposits, which are worm-free because of anthropogenic activities such as open-cast mining, soil sterilisation, consistent pollution or remediation of contaminated soil. Earthworms live in close interaction with their soil environment and are able to change it considerably by their burrowing and litter comminuting behaviour. While earthworms have been studied extensively, several questions still remain unanswered such as: What are the characteristics of successful early colonisers? Do they function well in dispersal, individual establishment or population growth? Do the negative environmental conditions in these kinds of anthropogenic soils hamper colonization or are these colonizers relatively resistant to it? To what extent does colonization change the characteristics of the colonized substrate? In short, do earthworms impact the soil? In this paper, the characteristics that make earthworms successful colonisers are briefly described as well as which species are the most successful and under what circumstances, and what do earthworms contribute to the total process of succession. We propose that it is not so much eco-type or r-K strategy that govern success and succession of earthworm colonisation but rather environmental flexibility not only towards pH, desiccation, and temperature but also towards contaminants such as heavy metals. Moreover, the formation of an organic litter layer, in close connection with re-vegetation of the area, is essential for establishing earthworm populations, which, at first, are mainly superficially and shallow active species. The burrowing and organic matter digesting activity of these earthworms changes the upper soil to a well mixed humus layer suitable for deep burrowing earthworm species.

The polyacrylamide gel electrophoresis technique was used for comparison of common protein of body wall tissues in five species of Lumbricidae (Eisenia nordenskioldi, E.fetida, Lumbricus rubellus, Aporrectodea caliginosa, A. longa). The statistic processing of indexes of electrophoretic similarity revealed three levels of similarity: intraspecies, interspecies, and intergenus. It was discovered that the similarity of E. nordenskioldi and E.fetida proteins corresponded to the intergenus level. The use of this method for determination of the earthworm's taxonomic status is discussed.

A Pb-mine site situated on acidic soil, but comprising of Ca-enriched islands around derelict buildings was used to study the spatial pattern of genetic diversity in Lumbricus rubellus. Two distinct genetic lineages ('A' and 'B'), differentiated at both the mitochondrial (mtDNA COII) and nuclear level (AFLPs) were revealed with a mean inter-lineage mtDNA sequence divergence of approximately 13%, indicative of a cryptic species complex. AFLP analysis indicates that lineage A individuals within one central 'ecological island' site are uniquely clustered, with little genetic overlap with lineage A individuals at the two peripheral sites. FTIR microspectroscopy of Pb-sequestering chloragocytes revealed different phosphate profiles in residents of adjacent acidic and calcareous islands. Bioinformatics found over-representation of Ca pathway genes in EST(Pb) libraries. Subsequent sequencing of a Ca-transport gene, SERCA, revealed mutations in the protein's cytosolic domain. We recommend the mandatory genotyping of all individuals prior to field-based ecotoxicological assays, particularly those using discriminating genomic technologies., (Copyright (c) 2009 Elsevier Ltd. All rights reserved.)

Opistocystidae Cernosvitov, 1936 is a largely Neotropical oligochaete taxon containing seven species. Its familial status has never been formally challenged, although possible close relationships with Naididae and Phreodrilidae have been noted. Mitochondrial 12S and 16S rDNA, and nuclear 18S rDNA, of a range of aquatic oligochaete taxa, including Trieminentia corderoi (Opistocystidae), were analysed by Bayesian inference. This showed that T. corderoi is a derived lineage within Naididae, closely related to Pristina and its monotypic subfamily Pristininae. Opistocystidae as a whole (with its three genera, Opistocysta, Trieminentia, and Crustipellis) is thus likely to be a group within Naididae.