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Three new aurantiactinomyxon types are described from the oligochaete Ilyodrilus templetoni (Southern, 1909) (Naididae) collected from a northern Portuguese estuary, based on light microscopy and sequencing of the 18S rDNA. The addition of I. templetoni to the group of freshwater annelids known to be permissive for aurantiactinomyxon development reinforces the crucial role of naidids in the evolution and settlement of myxozoans in estuarine environments. Maximum likelihood and Bayesian inference analyses of a comprehensive 18S rDNA dataset placed the novel types within the Paramyxidium clade. This positioning suggests them as probable life cycle counterparts to Paramyxidium spp. that most likely infect the European eel Anguilla anguilla, as the sole representative of Elopomorpha in Portuguese rivers. Although distance estimation revealed a genetic difference of only 0.4 % between Aurantiactinomyxon types 1 and 3, this value was determined to be representative of interspecific variability based on the consistent matching of both genotypes with distinct actinospore morphologies, and potential richness of closely related species of Paramyxidium infecting the European eel in Portuguese waters. The clustering of aurantiactinomyxon types within distinct myxosporean lineages, representative of the suborders Variisporina and Platysporina, demonstrates that the aurantiactinomyxon morphotype is highly functional in promoting myxozoan infections in estuarine environments., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Naidids are tiny, transparent freshwater oligochaetes, which are well known for their ability to propagate asexually. Despite the fact that sexually mature individuals and cocoons with embryos are sometimes found in nature, in long-period laboratory cultures, worms reproduce agametically only. In this paper, we showed, for the first time, the expression of Vasa , Piwi , and Pl10 homologs in mature Pristina longiseta worms with well-developed reproductive system structures and germ cells. Although the animals have been propagated asexually by paratomic fission for over 20 years in our lab, some individuals become sexualized under standard conditions for our laboratory culture and demonstrate various stages of maturation. The fully matured animals developed a complete set of sexual apparatus including spermatheca, atrium, seminal vesicles, and ovisac. They also had a clitellum and were able to form cocoons. The cues for the initiation of sexual maturation are still unknown for P. longiseta ; nevertheless, our data suggest that the laboratory strain of P. longiseta maintains the ability to become fully sexually mature and to establish germline products even after a long period of agametic reproduction. On the other hand, many of the sexualized worms formed a fission zone and continued to reproduce asexually. Thus, in this species, the processes of asexual reproduction and sexual maturation do not preclude each other, and Vasa , Piwi , and Pl10 homologs are expressed in both somatic and germline tissue including the posterior growth zone, fission zone, nervous system, germline cells, and gametes.

Aquatic worms are considered a suitable group to evaluate the effects of contaminants on the environment, although one of the main challenges is to use the species of local occurrence. Recently, Pristina longiseta was suggested to be used in acute bioassays. In this context, this study aimed to establish a chronic exposure for ecotoxicological bioassays using the cosmopolitan species of occurrence in Brazilian freshwater P. longiseta. Firstly, we tested three exposure times (4, 7, and 10 days) under the presence or absence of aeration for reproduction outputs. After determining the best configuration (7 days without aeration), we assessed the effects of the chronic exposures using the standardized reference substance potassium chloride (KCl), the antibiotic sulfamethoxazole (SMX), the flame retardant tetrabromobisphenol A (TBBPA), and the sugarcane vinasse. Our results showed suitability for applying the chronic exposure using P. longiseta and indicated the sensitivity of the offspring to KCl (EC50-7d = 0.51 g/L). Sulfamethoxazole and TBBPA caused a significant decrease in the offspring of P. longiseta (EC50-7d = 59.9 µg/L and < 62.5 µg/L, respectively). Sugarcane vinasse showed high toxicity for the species, and 4.26% of vinasse was calculated as EC50-7d. Therefore, the described protocol was successfully applied as an ecotoxicological bioassay to evaluate the effects of environmental contaminants on the reproduction rate of the freshwater worm P. longiseta., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Exploring the effectiveness of DNA barcoding in species identification is a prerequisite for biodiversity conservation and environmental monitoring. Aquatic oligochaetes could serve as excellent indicators in aquatic monitoring programmes. However, few studies have examined the effectiveness of DNA barcoding in these specific organisms. The mitochondrial cytochrome C oxidase (COI) gene of 83 specimens belonging to 40 species of 18 genera were sequenced in this study. The results showed that there was a barcode gap between species of Naididae and the intraspecific genetic distances of each species were smaller than interspecific genetic distances. The classification results of ABGD (Automatic Barcode Gap Discovery) were consistent with those of morphological identification, except for Tubifextubifex and Lumbriculusvariegatus . All species were successfully distinguished in the phylogenetic tree, based on the ITS2 region, which was coincident with the morphological result. Our results provided evidence that DNA barcoding can be used as an effective and convenient tool for species identification of the family Naididae and even for other aquatic oligochaetes., (Tingting Zhou, Wei Jiang, Hongzhu Wang, Yongde Cui.)

Ecotoxicological bioassays have been widely utilized to evaluate the toxicity of substances to organisms. However, the main challenge for researchers is finding native species to assess the effects of pollutants on aquatic biota. The tropical Oligochaeta, Allonais inaequalis, can be used as a test organism in bioassays to understand the effects of toxicants on aquatic ecosystems and their impact on native aquatic biota. In this study, we tested four methodological designs to validate the use of our "Allonais inaequalis reproduction test" as an ecotoxicological bioassay. For each sample, the assay consisted of a bottle containing 10 mg of sterilized fine sand, 60 mL of dechlorinated tap water and 6 organisms, fed at the beginning of the test and again after 5 days. The assay was first established in a controlled environment and then used to evaluate a stressed environment containing one of the following three toxicants suggested by the OECD (2008) and Corbi et al. (2015): zinc chloride, copper sulfate, or potassium chloride. Our results showed that the best experimental design for reproduction analysis was a static, long-term bioassay, which lasted 10 days without aeration and allowed for the reproduction of multiple generations (10 ± 5 new organisms). The observed inhibition reproduction by toxicants (EC50 ranging between 0.2 mg L -1 and 1.36 g L -1 ) validated the methods used in this paper. The use of a reproduction endpoint is a new contribution to the ecotoxicological toolbox, examining responses from a native organism to predict the effects of pollutants in an aquatic environment.

A new species of naidid oligochaete, Dero rwandae, detected in the bladder and the Wolffian ducts of reed frogs Hyperolius kivuensis from Rwanda, is described. Until now, D. bauchiensis was the only endoparasitic Dero known to infect African frogs infesting the eyes and Harderian glands. To the best of our knowledge, the finding of D. rwandae is the first record of an African Dero species infecting the urinary tract of anurans. In general morphology, the two African Dero parasites resemble each other, but differences in the features of ventral setae morphology exist. Parts of the mitochondrial 16S rRNA locus and the nuclear 18S and 28S rRNA loci were sequenced to assess the phylogenetic relationships to other Dero spp. Among those few species, that are barcoded so far, the closest relative of the new taxon is D. superterrenus, a free-living South American species. The species groups formerly termed subgenera Allodero, Aulophorus and Dero within the genus Dero do not represent distinct evolutionary lineages and the genus is paraphyletic including Branchiodrilus.

We investigated the involvement of oligochaetes in the life cycles of fresh water myxozoan parasites in Brazil. In a fish farm in the State of Mato Grosso do Sul, we examined 192 oligochaetes and found that two (1%) released Aurantiactinomyxon type actinospores. We identified infected oligochaetes by morphology: both were Pristina synclites, from family Naididae. This is the first report of the involvement of this species in the life cycle of myxozoans. Small-subunit ribosomal DNA sequences of Aurantiactinomyxon type 1 (1882 nt) and Aurantiactinomyxon type 2 (1900 nt) did not match any previously sequenced myxozoan in the NCBI database, with the highest BLAST search similarities of 83% with Myxobolus batalhensis MF361090 and 93% with Henneguya maculosus KF296344, respectively, and the two aurantiactinomyxons were only 75% similar to each other (over ~ 1900 bases). Phylogenetic analyses showed that Aurantiactinomyxon type 1 had closest affinities with myxozoans from fish hosts in Order Characiformes, and Aurantiactinomyxon type 2 had affinities with myxozoans from fish of Order Siluriformes.

The phylogenetic relationships within Naidinae (Annelida, Clitellata, Naididae) were investigated, using six molecular markers, both mitochondrial (12S rDNA, 16S rDNA, the COI gene) and nuclear (18S rDNA, 28S rDNA, the ITS region). Thirty-seven nominal species, representing 16 of the 22 genera recognized in the subfamily, were included, and the Nais communis/variabilis species complex was represented by six different morphotypes. Ten other species of Naididae were selected as outgroups. The data were analysed by Bayesian inference and Maximum Likelihood. The phylogeny corroborates monophyly of the Naidinae, and the separate status of the genus Pristina (Pristininae) and the Opistocystinae. Relationships within Naidinae are largely well supported, but in some parts unexpected: (1) A clade containing the largely tropical genera Dero and Branchiodrilus is sister to the rest of the subfamily, and together with a third tropical genus, Allonais, they form a basal paraphyly. All these genera show morphological adaptations to environmental hypoxia, leading to the conclusion that Naidinae originated in tropical freshwaters. (2) The genera Dero, Nais and Piguetiella are paraphyletic. (3) At least Branchiodrilus, Paranais, Chaetogaster, Nais, Stylaria appear to contain cryptic species. Morphological characters, especially those associated with chaetae, are to a great extent homoplastic within Naidinae, which certainly has contributed to the overall taxonomic confusion of this subfamily., (Copyright © 2017 Elsevier Inc. All rights reserved.)

One of the goals of urban ecology is to link community structure to ecosystem function in urban habitats. Pollution-tolerant wetland invertebrates have been shown to enhance greenhouse gas (GHG) flux in controlled laboratory experiments, suggesting that they may influence urban wetland roles as sources or sinks of GHG. However, it is unclear if their effects can be detected in highly variable conditions in a field setting. Here we use an extensive data set on carbon dioxide (CO 2 ), methane (CH 4 ), and nitrous oxide (N 2 O) flux in sediment cores (n = 103) collected from 10 urban wetlands in Melbourne, Australia during summer and winter in order to test for invertebrate enhancement of GHG flux. We detected significant multiplicative enhancement effects of temperature, sediment carbon content, and invertebrate density on CH 4 and CO 2 flux. Each doubling in density of oligochaete worms or large benthic invertebrates (oligochaete worms and midge larvae) corresponded to ~42% and ~15% increases in average CH 4 and CO 2 flux, respectively. However, despite exceptionally high densities, invertebrates did not appear to enhance N 2 O flux. This was likely due to fairly high organic carbon content in sediments (range 2.1-12.6%), and relatively low nitrate availability (median 1.96 μmol/L NO 3 - -N), which highlights the context-dependent nature of community structural effects on ecosystem function. The invertebrates enhancing GHG flux in this study are ubiquitous, and frequently dominate faunal communities in impaired aquatic ecosystems. Therefore, invertebrate effects on CO 2 and CH 4 flux may be common in wetlands impacted by urbanization, and urban wetlands may make greater contributions to the total GHG budgets of cities if the negative impacts of urbanization on wetlands are left unchecked., (© 2017 by the Ecological Society of America.)

Introduction: An important goal for understanding how animals have evolved is to reconstruct the ancestral features and evolution of the nervous system. Many inferences about nervous system evolution are weak because of sparse taxonomic sampling and deep phylogenetic distances among species compared. Increasing sampling within clades can strengthen inferences by revealing which features are conserved and which are variable within them. Among the Annelida, the segmented worms, the Clitellata are typically considered as having a largely conserved neural architecture, though this view is based on limited sampling., Results: To gain better understanding of nervous system evolution within Clitellata, we used immunohistochemistry and confocal laser scanning microscopy to describe the nervous system architecture of 12 species of the basally branching family Naididae. Although we found considerable similarity in the nervous system architecture of naidids and that of other clitellate groups, our study identified a number of features that are variable within this family, including some that are variable even among relatively closely related species. Variable features include the position of the brain, the number of ciliary sense organs, the presence of septate ventral nerve cord ganglia, the distribution of serotonergic cells in the brain and ventral ganglia, and the number of peripheral segmental nerves., Conclusions: Our analysis of patterns of serotonin immunoreactive perikarya in the central nervous system indicates that segmental units are not structurally homogeneous, and preliminary homology assessments suggest that whole sets of serotonin immunoreactive cells have been gained and lost across the Clitellata. We also found that the relative position of neuroectodermal and mesodermal segmental components is surprisingly evolutionarily labile; in turn, this revealed that scoring segmental nerves by their position relative to segmental ganglia rather than to segmental septa clarifies their homologies across Annelida. We conclude that fine taxonomic sampling in comparative studies aimed at elucidating the evolution of morphological diversity is fundamental for proper assessment of trait variability.

One new species of Naidinae (Oligochaeta, Naididae), Naisbadia sp. n. and one new record species from China, Tubifexmontanus Kowalewski, 1919 (Tubificinae) are found in southern Tibet. The new species is distinguished from congeners by its large area of reddish brown pigment in the anterior segments I-VIII, serrate hairs, pectinate needles with 1-2 intermediate teeth, ventral chaetae partly with 1-2 fine intermediate teeth and wave-like movements. The new material of the species Tubifexmontanus differs slightly from the previous descriptions by its vas deferens entering atrium subapically, wide ental end of penial sheath and smooth hair chaetae.

Three new species of Potamothrix Vejdovský & Mrázek, 1902 (Oligochaeta: Tubificinae), Potamothrix praeprostatussp. n., Potamothrix paramoldaviensissp. n. and Potamothrix parabedotisp. n., are reported from Fuxian Lake of Yunnan Province, Southwest China. Potamothrix praeprostatusdiffers from its allies by its prostate glands joining atria in its proximal to middle portion, and spermathecal chaetae. Potamothrix paramoldaviensis is distinguishable from its allies by having penial chaeta but no penes, and differs from Potamothrix moldaviensisby its homogenous atrium. Potamothrix parabedoti is distinctive in the position of its reproductive organs, and differs from Potamothrix bedoti by its homogenous atrium. Hitherto, 34 freshwater oligochaete species have been recorded in Yunnan Province, including nine endemic species from the plateau lakes.