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Microturbellarian flatworms comprise a diverse assemblage amongst meiofauna. These animals primarily exhibit carnivorous feeding habits, preying on various organisms, such as crustaceans, annelids and even other microturbellarians. However, details of their diet are poorly known. This study represents the first documentation of a proseriate preying upon a rhabdocoel. The proseriate was extracted from the sediment and studied alive. Within its digestive tract, structures of the reproductive systems of its prey were observed and identified as belonging to Phonorhynchopsishaegheni , a predatory turbellarian as well. No remains of any other organisms were detected. This finding underscores the relevance of Proseriata as top-level predators within the meiofaunal trophic web, a role that warrants further consideration beyond what has been previously acknowledged., Competing Interests: No conflict of interest to declare Disclaimer: This article is (co-)authored by any of the Editors-in-Chief, Managing Editors or their deputies in this journal., (Claudia Sanjuan Hernández, Marco Curini-Galletti, Marlies Monnens, Tom Artois, Yander L. Diez.)

Invasive Nile tilapias negatively impact native tilapia species through hybridisation and competition. However, the co-introduction of parasites with Nile tilapia, and subsequent changes in parasite communities, are scarcely documented. Monogeneans are known pathogens of cultured Nile tilapia, although little is known about their fate once Nile tilapias establish in new ecosystems. We investigate the parasitological consequences of Nile tilapia introduction on native tilapias in basins in Cameroon, the Democratic Republic of the Congo (DRC), and Zimbabwe, focusing on ectoparasitic dactylogyrids (Monogenea). Using the mitochondrial cytochrome oxidase c subunit I (COI) and nuclear 18S-internal transcribed spacer 1 (18S-ITS1) rDNA region of 128 and 166 worms, respectively, we evaluated transmission of several dactylogyrid species. Parasite spillover from Nile tilapia was detected for Cichlidogyrus tilapiae to Coptodon guineensis in Cameroon, Cichlidogyrus thurstonae to Oreochromis macrochir in the DRC, and Cichlidogyrus halli and C. tilapiae to Coptodon rendalli in Zimbabwe. Parasite spillback to Nile tilapia was detected for Cichlidogyrus papernastrema and Scutogyrus gravivaginus from Tilapia sparrmanii and Cichlidogyrus dossoui from C. rendalli or T. sparrmanii in the DRC, and Cichlidogyrus chloeae from Oreochromis cf. mortimeri and S. gravivaginus from O. macrochir in Zimbabwe. 'Hidden' transmissions (i.e. transmission of certain parasite lineages of species that are naturally present on both alien and native hosts) were detected for C. tilapiae and Scutogyrus longicornis between Nile tilapia and Oreochromis aureus and C. tilapiae between Nile tilapia and Oreochromis mweruensis in the DRC, and Cichlidogyrus sclerosus and C. tilapiae between Nile tilapia and O. cf. mortimeri in Zimbabwe. A high density of Nile tilapia occurring together with native tilapias, and the broad host range and/or environmental tolerance of the transmitted parasites, are proposed as factors behind parasite transmission through ecological fitting. However, continuous monitoring and the inclusion of environmental variables are necessary to understand the long-term consequences of these transmissions on native tilapias and to elucidate other underlying factors influencing these transmissions., (Copyright © 2023 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Clinostomum complanatum (Rudolphi, 1814) is an economically important parasitic flatworm (Trematoda, Digenea), yet little is known on the population structure of these animals. We characterise a new mitochondrial genome for C. complanatum, derived from an Iranian specimen. The newly obtained sequence is used to position the species in the digenean tree of life. The first-ever intraspecific comparison at mitogenome scale within C. complanatum revealed a high degree of similarity to the previously sequenced mitogenome of a distant (Italian) population. Avian migratory routes mirror phylogenetic clustering, and hence we suggest that infection of a flying host enables genetic exchange between parasites across large geographic distances. Comparative mitogenomic work in Clinostomum spp. at both the intra- (C. complanatum) and interspecific (C. complanatum-C. sinensis) level further shows that usage of new and/or additional mitochondrial markers is preferred over single-gene methods for high-resolution diagnostics and population biology., 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 © 2022 Elsevier B.V. All rights reserved.)

Urbanisation has become one of the major anthropogenic drivers behind insect decline in abundance, biomass and species richness over the past decades. As a result, bees and other pollinators' natural habitats are reduced and degraded. Green roofs are frequently recommended as ways to counter the negative impacts of urbanisation on nature and enhance the amount of green space in cities. In this study we evaluated the pollinator (more specifically wild bees and hoverflies) diversity, abundance and species richness on twenty green roofs in Antwerp, Belgium. We analysed the influence of roof characteristics (age, surface area, height, percent cover of green space surrounding each site) on species richness or abundance of pollinators. In total we found 40 different wild bee species on the green roofs. None of the physical roof characteristics appear to explain differences in wild bees species richness and abundance. Neither could we attribute the difference in roof vegetation cover, i.e. roofs build-up with only Sedum species and roofs with a combined cover of Sedum, herbs and grasses, to differences in diversity, abundance, or species richness. We found a positive trend, although not significant, in community weighted mean body size for wild bees with an increase in green roof surface area. Roof wild bee communities were identified as social polylectic individuals, with a preference for ground nesting. Only eleven individuals from eight different hoverfly species were found. Our results show that green roofs can be a suitable habitat for wild bee species living in urban areas regardless of the roofs' characteristics, but hoverflies have more difficulties conquering these urban green spaces., (© 2023. The Author(s).)

Imaging of living animals allows the study of metabolic processes in relation to cellular structures or larger functional entities. To enable in vivo imaging during long-term time-lapses in planarians, we combined and optimized existing protocols, resulting in an easily reproducible and inexpensive procedure. Immobilization with low-melting-point agarose eliminates the use of anesthetics, avoids interfering with the animal during imaging-functionally or physically-and allows recovering the organisms after the imaging procedure. As an example, we used the immobilization workflow to image the highly dynamic and fast-changing reactive oxygen species (ROS) in living animals. These reactive signaling molecules can only be studied in vivo and mapping their location and dynamics during different physiological conditions is crucial to understand their role in developmental processes and regeneration. In the current protocol, we describe both the immobilization and ROS detection procedure. We used the intensity of the signals together with pharmacological inhibitors to validate the signal specificity and to distinguish it from the autofluorescent nature of the planarian., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

A substantial portion of biodiversity has evolved through adaptive radiation. However, the effects of explosive speciation on species interactions remain poorly understood. Metazoan parasites infecting radiating host lineages could improve our knowledge because of their intimate host relationships. Yet limited molecular, phenotypic and ecological data discourage multivariate analyses of evolutionary patterns and encourage the use of discrete characters. Here, we assemble new molecular, morphological and host range data widely inferred from a species-rich lineage of parasites (Cichlidogyrus, Platyhelminthes: Monogenea) infecting cichlid fishes to address data scarcity. We infer a multimarker (28S/18S rDNA, ITS1, COI mtDNA) phylogeny of 58 of 137 species and characterize major lineages through synapomorphies inferred from mapping morphological characters. We predict the phylogenetic position of species without DNA data through shared character states, a morphological phylogenetic analysis, and a classification analysis with support vector machines. Based on these predictions and a cluster analysis, we assess the systematic informativeness of continuous characters, search for continuous equivalents for discrete characters, and suggest new characters for morphological traits not analysed to date. We also model the attachment/reproductive organ and host range evolution using the data for 136 of 137 described species and multivariate phylogenetic comparative methods (PCMs). We show that discrete characters not only can mask phylogenetic signals, but also are key for characterizing species groups. Regarding the attachment organ morphology, a divergent evolutionary regime for at least one lineage was detected and a limited morphological variation indicates host and environmental parameters affecting its evolution. However, moderate success in predicting phylogenetic positions, and a low systematic informativeness and high multicollinearity of morphological characters call for a revaluation of characters included in species characterizations., (© 2022 Willi Hennig Society.)

During colonial times, Nile tilapia Oreochromis niloticus (Linnaeus, 1758) was introduced into non-native parts of the Congo Basin (Democratic Republic of the Congo, DRC) for the first time. Currently, it is the most farmed cichlid in the DRC, and is present throughout the Congo Basin. Although Nile tilapia has been reported as an invasive species, documentation of historical introductions into this basin and its consequences are scant. Here, we study the genetic consequences of these introductions by genotyping 213 Nile tilapia from native and introduced regions, focusing on the Congo Basin. Additionally, 48 specimens from 16 other tilapia species were included to test for hybridization. Using RAD sequencing (27,611 single nucleotide polymorphisms), we discovered genetic admixture with other tilapia species in several morphologically identified Nile tilapia from the Congo Basin, reflecting their ability to interbreed and the potential threat they pose to the genetic integrity of native tilapias. Nile tilapia populations from the Upper Congo and those from the Middle-Lower Congo are strongly differentiated. The former show genetic similarity to Nile tilapia from the White Nile, while specimens from the Benue Basin and Lake Kariba are similar to Nile tilapia from the Middle-Lower Congo, suggesting independent introductions using different sources. We conclude that the presence of Nile tilapia in the Congo Basin results from independent introductions, reflecting the dynamic aquaculture history, and that their introduction probably leads to genetic interactions with native tilapias, which could lower their fitness. We therefore urge avoiding further introductions of Nile tilapia in non-native regions and to use native tilapias in future aquaculture efforts., (© 2022 John Wiley & Sons Ltd.)

Dactylogyridae is one of the most studied families of parasitic flatworms with more than 1000 species and 166 genera described to date including ecto- and endoparasites. Dactylogyrid monogeneans were suggested as model organisms for host-parasite macroevolutionary and biogeographical studies due to the scientific and economic importance of some of their host lineages. Consequently, an array of phylogenetic research into different dactylogyrid lineages has been produced over the past years but the last family-wide study was published 16 years ago. Here, we provide a meta-analysis of the phylogenetic relationships of Dactylogyridae including representatives of all genera with available molecular data (n = 67). First, we investigate the systematic informativeness of morphological characters widely used to diagnose dactylogyrid genera through a parsimony analysis of the characters, character mapping, and phylogenetic comparative methods. Second, we provide an overview of the current state of the systematics of the family and its subfamilies, and summarise potentially poly- and paraphyletic genera. Third, we elaborate on the implications of taxonomic, citation, and confirmation bias in past studies. Fourth, we discuss host range, biogeographical, and freshwater-marine patterns. We found two well-supported macroclades which we assigned to the subfamilies Dactylogyrinae and Ancyrocephalinae. These subfamilies further include 16 well-supported clades with only a few synapomorphies that could be deduced from generic diagnoses in the literature. Furthermore, few morphological characters considered systematically informative at the genus level display a strong phylogenetic signal. However, the parsimony analysis suggests that these characters provide little information on the relationships between genera. We conclude that a strong taxonomic bias and low coverage of DNA sequences and regions limit knowledge on morphological and biogeographical evolutionary patterns that can be inferred from these results. We propose addressing potential citation and confirmation biases through a 'level playing field' multiple sequence alignment as provided by this study., (Copyright © 2022. Published by Elsevier Ltd.)

The origin of introduced Nile tilapia stocks in sub-Saharan Africa is largely unknown. In this study, the potential of monogeneans as a biological tag and magnifying glass is tested to reveal their hosts' stocking history. The monogenean gill community of different Nile tilapia populations in sub-Saharan Africa was explored, and a phylogeographic analysis was performed based on the mitogenomes of four dactylogyrid species (Cichlidogyrus halli, C. sclerosus, C. thurstonae, and Scutogyrus longicornis). Our results encourage the use of dactylogyrids as biological tags. The magnifying glass hypothesis is only confirmed for C. thurstonae, highlighting the importance of the absence of other potential hosts as prerequisites for a parasite to act as a magnifying glass. With the data generated here, we are the first to extract mitogenomes from individual monogeneans and to perform an upscaled survey of the comparative phylogeography of several monogenean species with unprecedented diagnostic resolution., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

During their annual migration, avian migrants alternate stopover periods, for refuelling, with migratory flight bouts. We hypothesise that European Nightjars (Caprimulgus europaeus) adapt their daily migration tactics in association with biomes. We tracked the autumn migration of 24 European Nightjars, from breeding populations in Mongolia, Belgium and UK, using GPS-loggers and multi-sensor data loggers. We quantified crepuscular and nocturnal migration and foraging probabilities, as well as daily travel speed and flight altitude during active migration in response to biomes. Nightjars adopt a rush tactic, reflected in high daily travel speed, flight altitude and high migration probabilities at dusk and at night, when travelling through ecological barriers. Migration is slower in semi-open, hospitable biomes. This is reflected in high foraging probabilities at dusk, lower daily travel speed and lower migration probabilities at dusk. Our study shows how nightjars switch migration tactics during autumn migration, and suggest nightjars alternate between feeding and short migratory flight bouts within the same night when travelling through suitable habitats. How this may affect individuals' fuel stores and whether different biomes provide refuelling opportunities en route remains to be investigated, to understand how future land-use change may affect migration patterns and survival probabilities., (© 2022. The Author(s).)

We describe a new genus of dalytyphloplanid rhabdocoels, with seven new species. Orostylis gen. nov. has a unique combination of characters including the presence of a sclerotised stylet, an anteriorly positioned male copulatory organ with the male genital pore in the buccal cavity, and the absence of an oviduct. The ovary empties directly into the intestinal lumen. Orostylis dohae sp. nov., Orostylis asinaraensis sp. nov., Orostylis caecus sp. nov., Orostylis distortus sp. nov., Orostylis donanae sp. nov., Orostylis gallicus sp. nov., and Orostylis timucuorum sp. nov. are distinguished from each other by the structure of the sclerotised parts of the male copulatory organ. Molecular and morphological evidence place the new genus in Neodalyellida Willems et al. 2006, and similarities with other taxa are discussed.

Unlike their marine counterparts, tropical freshwater clupeids receive little scientific attention. However, they sustain important fisheries that may be of (inter)national commercial interest. Africa harbours over 20 freshwater clupeid species within Pellonulini. Recent research suggests their most abundant parasites are gill-infecting monogenean flatworms within Kapentagyrus . After inspecting specimens of 12 freshwater clupeids from West and Central Africa, mainly sourced in biodiversity collections, we propose 11 new species of Kapentagyrus, which we describe using their haptoral and genital morphology. Because of their high morphological similarity, species delineation relies mostly on the morphometrics of anchors and hooks. Specifically, earlier, molecular taxonomic work indicated that the proportion between the length of the anchor roots, and between the hook and anchor length, is diagnostic. On average, about one species of Kapentagyrus exists per pellonuline species, although Pellonula leonensis harbours four species and Microthrissa congica two, while Microthrissa moeruensis and Potamothrissa acutirostris share a gill monogenean species. This study more than quadruples the number of known species of Kapentagyrus , also almost quadrupling the number of pellonuline species of which monogeneans are known. Since members of Kapentagyrus are informative about their hosts' ecology, evolutionary history, and introduction routes, this enables a parasitological perspective on several data-poor African fisheries.

Little phylogeographic structure is presumed for highly mobile species in pelagic zones. Lake Tanganyika is a unique ecosystem with a speciose and largely endemic fauna famous for its remarkable evolutionary history. In bathybatine cichlid fishes, the pattern of lake-wide population differentiation differs among species. We assessed the congruence between the phylogeographic structure of bathybatine cichlids and their parasitic flatworm Cichlidogyrus casuarinus to test the magnifying glass hypothesis. Additionally, we evaluated the use of a PoolSeq approach to study intraspecific variation in dactylogyrid monogeneans. The lake-wide population structure of C. casuarinus ex Hemibates stenosoma was assessed based on a portion of the cox 1 gene combined with morphological characterisation. Additionally, intraspecific mitogenomic variation among 80 parasite samples from one spatially constrained metapopulation was assessed using shotgun NGS. While no clear geographic genetic structure was detected in parasites, both geographic and host-related phenotypic variation was apparent. The incongruence with the genetic north-south gradient observed in H. stenosoma may be explained by the broad host range of this flatworm including eupelagic bathybatine host species that form panmictic populations across the lake. In addition, we present the first parasite mitogenome from Lake Tanganyika and propose a methodological framework for studying the intraspecific mitogenomic variation of dactylogyrid monogeneans.

A comprehensive morphological and taxonomic account of the members of the genus Cheliplana de Beauchamp, 1927 is presented. Six new species are described: Cheliplana asinaraensis n. sp., C. cubana n. sp., C. curacaoensis n. sp., C. hawaiiensis n. sp., C. longissima n. sp. and C. mauii n. sp. The new species are mainly distinguished from each other and from other representatives of Cheliplana by the organisation of the reproductive system and the structure of the cirrus. Furthermore, C. triductibus Van Steenkiste, Volonterio, Schockaert Artois, 2008 is considered a junior synonym of Cheliplana deverticula Ax, 2008. The two subspecies of Cheliplana asica Marcus, 1952, C. asica asica and C. asica terminalis Brunet, 1968, are considered separate species. The systematic position of the genus Dactyloplana Armonies, 2018 is discussed, and its synonymy with Cheliplana is retained. As such, this brings the total number of species of Cheliplana to 49. Finally, we provide an identification key to the members of the genus, based on characters that enable identification to species level in the field.

A strict coordination between pro- and antioxidative molecules is needed for normal animal physiology, although their exact function and dynamics during regeneration and development remains largely unknown. Via in vivo imaging, we were able to locate and discriminate between reactive oxygen species (ROS) in real-time during different physiological stages of the highly regenerative planarian Schmidtea mediterranea . All ROS signals were strong enough to overcome the detected autofluorescence. Combined with an in situ characterisation and quantification of the transcription of several antioxidant genes, our data showed that the planarian gut and epidermis have a well-equipped redox system. Pharmacological inhibition or RNA interference of either side of the redox balance resulted in alterations in the regeneration process, characterised by decreased blastema sizes and delayed neurodevelopment, thereby affecting tails more than heads. Focusing on glutathione, a central component in the redox balance, we found that it is highly present in planarians and that a significant reduction in glutathione content led to regenerative failure with tissue lesions, characterised by underlying stem cell alterations. This exploratory study indicates that ROS and antioxidants are tightly intertwined and should be studied as a whole to fully comprehend the function of the redox balance in animal physiology.

Parasite diversity above the Arctic circle remains understudied even for commercially valuable host taxa. Thorny skate, Amblyraja radiata, is a common bycatch species with a growing commercial value. Its natural range covers both sides of the North Atlantic including the Arctic zone. Svalbard is a Norwegian archipelago located on the northwest corner of the Barents Shelf which sustains a spectacular species diversity. So far, several monogenean species have been reported infecting thorny skate across the Atlantic Ocean. In the present study, we intend to fill in the knowledge gap on monogenean parasites infecting thorny skate in the northern part of its range and thus indirectly assess the connectivity between the thorny skate populations off the Svalbard coast and from previously studied locations. 46 monogenean individuals were recovered from 11 specimens of thorny skate. Following morphological and molecular assessment, two species of monogeneans, Acanthocotyle verrilli and Rajonchocotyle emarginata, were identified. The results serve as the northernmost record for both parasite genera and the first record of monogenean species off Svalbard. Detailed morphometric evaluation revealed a relatively high level of morphological variation in A. verrilli compared to its congeners. Phylogenetic reconstruction placed A. verrilli in a well-supported clade with A. imo. Our study also suggests high diagnostic significance of sclerotised structures in the identification of Rajonchocotyle. Even though the occurrence of two directly transmitted parasite species supports the previously suggested long-distance migration of A. radiata, future studies employing highly variable genetic markers are needed to assess the ongoing and historical migration patterns., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The taxon Koinocystididae is the third most species-rich family within Eukalyptorhynchia. However, its diversity and phylogeny have been largely neglected in former studies. We introduce three new genera and twelve new species of Koinocystididae including Simplexcystis asymmetrica gen. n. sp. n., Galapagetula cubensis sp. n., eight species of Reinhardorhynchus gen. n. and two species of Itaipusa. This raises the total number of species within Koinocystididae from 51 to 63. We also report on new distribution records for six known species: I. divae (Cuba, Panama and New Caledonia), I. karlingi (Sardinia and Lanzarote), Reinhardorhynchus riegeri comb. n. (Cuba), R. ruffinjonesi comb. n. (Cuba and Panama), Utelga heinckei (Cuba and Lanzarote), and U. pseudoheinckei (Sardinia). Simplexcystis asymmetrica gen. n. sp. n. is characterised by a male duct running eccentrically through the copulatory bulb, lack of any hard structures in the male system, lack of a bursa, and the fact that the epithelia of the female, the male, and part of the common atrium are covered by a brush border. Galapagetula cubensis sp. n. has a caudal gonopore, a divisa-type copulatory bulb with an unarmed penis papilla, and a female duct without a sphincter. The new species of Itaipusa and Reinhardorhynchus gen. n. differ from their congeners in the detailed structure of the copulatory bulb and especially the hard structures associated with it. In a molecular phylogenetic analysis based on all available 18S and 28S rDNA sequences of koinocystidids, we found support for the monophyly of the family and the genus Utelga Marcus, 1949. The genus Itaipusa is not monophyletic in that I. sinensis forms a clade with Rhinolasius dillonicus, while other species of Itaipusa that have a copulatory bulb armed with hooks form a clade together with Sekerana stolzi. As the type species of Itaipusa (I. divae) is in neither of these clades, we erected a new genus for I. sinensis (Koinogladius gen. n.) and one for species of Itaipusa having a hook-bearing copulatory bulb (Reinhardorhynchus gen. n.), respectively. Whether the remaining species of Itaipusa form a monophylum remains uncertain.

Silver nanoparticles (AgNPs) are widely incorporated in household, consumer and medical products. Their unintentional release via wastewaters raises concerns on their environmental impact, particularly for aquatic organisms and their associated bacterial communities. It is known that the microbiome plays an important role in its host's health and physiology, e.g. by producing essential nutrients and providing protection against pathogens. A thorough understanding of the effects of AgNPs on bacterial communities and on their interactions with the host is crucial to fully assess AgNP toxicity on aquatic organisms. Our results indicate that the microbiome of the invertebrate Schmidtea mediterranea, a freshwater planarian, is affected by AgNP exposure at the tested 10 μg/ml concentration. Using targeted amplification of the bacterial 16S rRNA gene V3-V4 region, two independent experiments on the microbiomes of adult worms revealed a consistent decrease in Betaproteobacteriales after AgNP exposure, mainly attributed to a decrease in Curvibacter and Undibacterium. Although developing tissues and organisms are known to be more sensitive to toxic compounds, three independent experiments in regenerating worms showed a less pronounced effect of AgNP exposure on the microbiome, possibly because underlying bacterial community changes during development mask the AgNP induced effect. The presence of a polyvinyl-pyrrolidone (PVP) coating did not significantly alter the outcome of the experiments compared to those with uncoated particles. The observed variation between the different experiments underlines the highly variable nature of microbiomes and emphasises the need to repeat microbiome experiments, within and between physiological states of the animal., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The first three mitochondrial (mt) genomes of endosymbiotic turbellarian flatworms are characterised for the rhabdocoels Graffilla buccinicola, Syndesmis echinorum and S. kurakaikina. Interspecific comparison of the three newly obtained sequences and the only previously characterised rhabdocoel, the free-living species Bothromesostoma personatum, reveals high mt genomic variability, including numerous rearrangements. The first intrageneric comparison within rhabdocoels shows that gene order is not fully conserved even between congeneric species. Atp8, until recently assumed absent in flatworms, was putatively annotated in two sequences. Selection pressure was tested in a phylogenetic framework and is shown to be significantly relaxed in this and another protein-coding gene: cox1. If present, atp8 appears highly derived in platyhelminths and its functionality needs to be addressed in future research. Our findings for the first time allude to a large degree of undiscovered (mt) genomic plasticity in rhabdocoels. It merits further attention whether this variation is correlated with a symbiotic lifestyle. Our results illustrate that this phenomenon is widespread in flatworms as a whole and not exclusive to the better-studied neodermatans., Competing Interests: Declaration of competing interest None., (Crown Copyright © 2020. Published by Elsevier B.V. All rights reserved.)

Resource distribution, predation risk and disturbance in space and time can affect how animals use their environment. To date few studies have assessed the spatiotemporal trade-off between resource acquisition and avoidance of risks and human disturbance in small protected areas embedded in an urban matrix. A better understanding of the forage-safety trade-off in urban protected areas (UPA) is key to the design of evidence-based approaches to deal with the ever-increasing human-wildlife impacts typical of UPA. Herein, we analyzed camera trap data to evaluate how two ungulate species trade fear for food in a 60 km 2 human-dominated UPA without natural predators. We found that wild boar (Sus scrofa) were predominantly active at night, while roe deer (Capreolus capreolus) showed a typical bimodal crepuscular activity pattern. Occupancy analysis indicated that deciduous forest and the presence of high seats for hunting played an important role in determining the space use of wild boar. For roe deer, we found indications that the presence of forest influenced space use, although the null model was retained among the top ranked models. Our results confirm that wild boar and roe deer are able to thrive in heavily human dominated landscapes characterized by intensive recreational use and hunting, such as protected areas embedded in an urban matrix., 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 © 2020 Elsevier B.V. All rights reserved.)

Given the global decline of many invertebrate food resources, it is fundamental to understand the dietary requirements of insectivores. We give new insights into the functional relationship between the spatial habitat use, food availability, and diet of a crepuscular aerial insectivore, the European Nightjar ( Caprimulgus europaeus ) by relating spatial use data with high-throughput sequencing (HTS) combined with DNA metabarcoding. Our study supports the predictions that nightjars collect a substantial part of their daily nourishment from foraging locations, sometimes at considerable distance from nesting sites. Lepidopterans comprise 65% of nightjars' food source. Nightjars tend to select larger species of Lepidoptera (>19 mm) which suggests that nightjars optimize the efficiency of foraging trips by selecting the most energetically favorable-larger-prey items. We anticipate that our findings may shed additional light on the interactions between invertebrate communities and higher trophic levels, which is required to understand the repercussions of changing food resources on individual- and population-level processes., Competing Interests: We declare we have no competing interests., (© 2020 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Gastrotricha and Platyhelminthes form a clade called Rouphozoa. Representatives of both taxa are main components of meiofaunal communities, but their role in the trophic ecology of marine and freshwater communities is not sufficiently studied. Traditional collection methods for meiofauna are optimized for Ecdysozoa, and include the use of fixatives or flotation techniques that are unsuitable for the preservation and identification of soft-bodied meiofauna. As a result, rouphozoans are usually underestimated in conventional biodiversity surveys and ecological studies. Here, we give an updated outline of their diversity and taxonomy, with some phylogenetic considerations. We describe successfully tested techniques for their recovery and study, and emphasize current knowledge on the ecology, distribution and dispersal of freshwater gastrotrichs and microturbellarians. We also discuss the opportunities and pitfalls of (meta)barcoding studies as a means of overcoming the taxonomic impediment. Finally, we discuss the importance of rouphozoans in aquatic ecosystems and provide future research directions to fill in crucial gaps in the biology of these organisms needed for understanding their basic role in the ecology of benthos and their place in the trophic networks linking micro-, meio- and macrofauna of freshwater ecosystems.

Biological rhythms of nearly all animals on earth are synchronized with natural light and are aligned to day-and-night transitions. Here, we test the hypothesis that the lunar cycle affects the nocturnal flight activity of European Nightjars ( Caprimulgus europaeus) . We describe daily activity patterns of individuals from three different countries across a wide geographic area, during two discrete periods in the annual cycle. Although the sample size for two of our study sites is small, the results are clear in that on average individual flight activity was strongly correlated with both local variation in day length and with the lunar cycle. We highlight the species' sensitivity to changes in ambient light and its flexibility to respond to such changes in different parts of the world., Competing Interests: We declare we have no competing interests., (© 2020 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Lake Tanganyika, East Africa, is the oldest and deepest African Great Lake and harbours one of the most diverse fish assemblages on earth. Two clupeid fishes, Limnothrissa miodon and Stolothrissa tanganicae, constitute a major part of the total fish catch, making them indispensable for local food security. Parasites have been proposed as indicators of stock structure in highly mobile pelagic hosts. We examined the monogeneans Kapentagyrus limnotrissae and Kapentagyrus tanganicanus (Dactylogyridae) infecting these clupeids to explore the parasites' lake-wide population structure and patterns of demographic history. Samples were collected at seven sites distributed across three sub-basins of the lake. Intraspecific morphological variation of the monogeneans (n = 380) was analysed using morphometrics and geomorphometrics of sclerotised structures. Genetic population structure of both parasite species (n = 246) was assessed based on a 415 bp fragment of the mitochondrial cytochrome c oxidase subunit I (COI) gene. Overall, we observed a lack of clear geographical morphological differentiation in both parasites along a north-south axis. This lack of geographical population structure was also reflected by a large proportion of shared haplotypes, and a pattern of seemingly unrestricted gene flow between populations. Significant morphological and genetic differentiation between some populations might reflect temporal differentiation rather than geographical isolation. Overall, the shallow population structure of both species of Kapentagyrus reflects the near-panmictic population structure of both host species as previously reported. Morphological differences related to host species identity of K. tanganicanus were consistent with incipient speciation at the genetic level. Both parasite species experienced a recent demographic expansion, which might be linked to paleohydrological events. Finally, interspecific hybridisation was found in Kapentagyrus, representing the first case in dactylogyrid monogeneans., (Copyright © 2020 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)

Background: Lake Tanganyika is considered a biodiversity hotspot with exceptional species richness and level of endemism. Given the global importance of the lake in the field of evolutionary biology, the understudied status of its parasite fauna is surprising with a single digenean species reported to date. Although the most famous group within the lake's fish fauna are cichlids, the pelagic zone is occupied mainly by endemic species of clupeids (Actinopterygii: Clupeidae) and lates perches (Actinopterygii: Latidae, Lates Cuvier), which are an important commercial source for local fisheries. In this study, we focused on the lake's four lates perches and targeted their thus far unexplored endoparasitic digenean fauna., Methods: A total of 85 lates perches from four localities in Lake Tanganyika were examined. Cryptogonimid digeneans were studied by means of morphological and molecular characterisation. Partial sequences of the nuclear 28S rRNA gene and the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene were sequenced for a representative subset of the specimens recovered. Phylogenetic analyses were conducted at the family level under Bayesian inference., Results: Our integrative approach revealed the presence of six species within the family Cryptogonimidae Ward, 1917. Three out of the four species of Lates were found to be infected with at least one cryptogonimid species. Two out of the three reported genera are new to science. Low interspecific but high intraspecific phenotypic and genetic diversity was found among Neocladocystis spp. Phylogenetic inference based on partial 28S rDNA sequences revealed a sister group relationship for two of the newly erected genera and their close relatedness to the widely distributed genus Acanthostomum Looss, 1899., Conclusions: The present study provides the first comprehensive characterisation of the digenean diversity in a fish family from Lake Tanganyika which will serve as a baseline for future explorations of the lake's digenean fauna. Our study highlights the importance of employing an integrative approach for revealing the diversity in this unique host-parasite system.

Pluripotent stem cells hold great potential for regenerative medicine. Increased replication and division, such is the case during regeneration, concomitantly increases the risk of adverse outcomes through the acquisition of mutations. Seeking for driving mechanisms of such outcomes, we challenged a pluripotent stem cell system during the tightly controlled regeneration process in the planarian Schmidtea mediterranea Exposure to the genotoxic compound methyl methanesulfonate (MMS) revealed that despite a similar DNA-damaging effect along the anteroposterior axis of intact animals, responses differed between anterior and posterior fragments after amputation. Stem cell proliferation and differentiation proceeded successfully in the amputated heads, leading to regeneration of missing tissues. Stem cells in the amputated tails showed decreased proliferation and differentiation capacity. As a result, tails could not regenerate. Interference with the body-axis-associated component β-catenin-1 increased regenerative success in tail fragments by stimulating proliferation at an early time point. Our results suggest that differences in the Wnt signalling gradient along the body axis modulate stem cell responses to MMS., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2020. Published by The Company of Biologists Ltd.)

Whereas most work to understand impacts of humans on biodiversity on coastal areas has focused on large, conspicuous organisms, we highlight effects of tourist access on the diversity of microscopic marine animals (meiofauna). We used a DNA metabarcoding approach with an iterative and phylogeny-based approach for the taxonomic assignment of meiofauna and relate diversity patterns to the numbers of tourists accessing sandy beaches on an otherwise un-impacted island National Park. Tourist frequentation, independently of differences in sediment granulometry, beach length, and other potential confounding factors, affected meiofaunal diversity in the shallow "swash" zone right at the mean water mark; the impacts declined with water depth (up to 2 m). The indicated negative effect on meiofauna may have a consequence on all the biota including the higher trophic levels. Thus, we claim that it is important to consider restricting access to beaches in touristic areas, in order to preserve biodiversity.

Background: Monogenea van Beneden, 1858 is a group of parasitic flatworms, commonly found infecting bony fish. Several genera, such as Cichlidogyrus Paperna, 1960, are reported to include potential pathogenic species that can negatively impact aquaculture fish stocks. They can switch from introduced to native fish and vice versa. In Africa (and all over the world), fish species belonging to Cichlidae are often kept in aquaculture and represent a major source of food. Thus, research on the biodiversity and occurrence of monogenean species on these fish is of importance for aquaculture and conservation. The present study is a survey of the diversity of species of Cichlidogyrus in the south of the Democratic Republic of the Congo (DRC) on three cichlid species: Orthochromis sp. 'Lomami', Serranochromis cf. macrocephalus, and Tilapia sparrmanii Smith, 1840., Methods: Specimens of Cichlidogyrus were isolated from the gills and mounted on glass slides with Hoyer's medium. The genital and haptoral hard parts were measured and drawn using interference contrast., Results: In total, six species of Cichlidogyrus were found, all new to science: C. bulbophallus n. sp. and C. pseudozambezensis n. sp. on S. cf. macrocephalus, C. flagellum n. sp. and C. lobus n. sp. on T. sparrmanii, C. ranula n. sp. on S. cf. macrocephalus and Orthochromis sp. 'Lomami', and C. maeander n. sp. found on Orthochromis sp. 'Lomami' and T. sparrmanii. The first four species are considered to be strict specialists, C. ranula n. sp. an intermediate generalist and C. maeander n. sp. a generalist. These parasite species show morphological similarities to species found in the Lower Guinea and Zambezi ichthyofaunal provinces, which might be explained by past river capture events between river systems of the Congo Province and both these regions., Conclusions: Serranochromis cf. macrocephalus and Orthochromis sp. 'Lomami' can harbour respectively three and two species of Cichlidogyrus, all described in this study. Tilapia sparrmanii can harbour seven species, of which three are described in the present study. These results highlight the species diversity of this parasite genus in the Congo Basin.

It is essential to gain knowledge about the causes and extent of migratory connectivity between stationary periods of migrants to further the understanding of processes affecting populations, and to allow efficient implementation of conservation efforts throughout the annual cycle. Avian migrants likely use optimal routes with respect to mode of locomotion, orientation and migration strategy, influenced by external factors such as wind and topography. In self-powered flapping flying birds, any increases in fuel loads are associated with added flight costs. Energy-minimizing migrants are therefore predicted to trade-off extended detours against reduced travel across ecological barriers with no or limited foraging opportunities. Here, we quantify the extent of detours taken by different populations of European nightjars Caprimulgus europaeus, to test our predictions that they used routes beneficial according to energetic principles and evaluate the effect of route shape on seasonal migratory connectivity. We combined data on birds tracked from breeding sites along a longitudinal gradient from England to Sweden. We analysed the migratory connectivity between breeding and main non-breeding sites, and en route stopover sites just south of the Sahara desert. We quantified each track's route extension relative to the direct route between breeding and wintering sites, respectively, and contrasted it to the potential detour derived from the barrier reduction along the track while accounting for potential wind effects. Nightjars extended their tracks from the direct route between breeding and main non-breeding sites as they crossed the Mediterranean Sea-Sahara desert, the major ecological barrier in the Palaearctic-African migration system. These clockwise detours were small for birds from eastern sites but increased from east to west breeding longitude. Routes of the tracked birds were associated with partial reduction in the barrier crossing resulting in a trade-off between route extension and barrier reduction, as expected in an energy-minimizing migrant. This study demonstrates how the costs of barrier crossings in prevailing winds can disrupt migratory routes towards slightly different goals, and thereby promote migratory connectivity. This is an important link between individual migration strategies in association with an ecological barrier, and both spatially and demographic population patterns., (© 2019 The Authors. Journal of Animal Ecology © 2019 British Ecological Society.)

In species richness studies, citizen-science surveys where participants make individual decisions regarding sampling strategies provide a cost-effective approach to collect a large amount of data. However, it is unclear to what extent the bias inherent to opportunistically collected samples may invalidate our inferences. Here, we compare spatial predictions of forest ground-floor bryophyte species richness in Limburg (Belgium), based on crowd- and expert-sourced data, where the latter are collected by adhering to a rigorous geographical randomisation and data collection protocol. We develop a log-Gaussian Cox process model to analyse the opportunistic sampling process of the crowd-sourced data and assess its sampling bias. We then fit two geostatistical Poisson models to both data-sets and compare the parameter estimates and species richness predictions. We find that the citizens had a higher propensity for locations that were close to their homes and environmentally more valuable. The estimated effects of ecological predictors and spatial species richness predictions differ strongly between the two geostatistical models. Unknown inconsistencies in the sampling process, such as unreported observer's effort, and the lack of a hypothesis-driven study protocol can lead to the occurrence of multiple sources of sampling bias, making it difficult, if not impossible, to provide reliable inferences.

Coelogynopora schockaerti n. sp. (Proseriata: Coelogynoporidae) is described from the northwestern Gulf of Lion, Mediterranean. Only a few representatives of this genus are reported from Mediterranean marine sandy shores and only from the northwestern to central Mediterranean, but all of these species also occur outside the Mediterranean. So far, C. schockaerti n. sp. is the only species exclusively known from the Mediterranean. The species is solely encountered in the Gulf of Lion, a coldwater pocket in the Mediterranean and can, therefore, be considered as a remnant of an ancient coldwater platyhelminth fauna. C. schockaerti n. sp. is characterized by the shape and size of the spines of the male copulatory organ and the presence, orientation, shape and size of the two accessory spines.  An identification key is given for the entire genus.

We report on the schizorhynch species collected in a survey in the eastern region of Cuba. Eighteen species were identified, of which only three are known to science: Cheliplana asica, C. terminalis, and Carcharodorhynchus flavidus. The 15 new species belong to three different genera: Cheliplana (five species), Carcharodorhynchus (four species), and Schizochilus (six species). The five new species of Cheliplana (C. gibarenha sp. n., C. santiaguera sp. n., C. spuriaseminalis sp. n., C. subproximalis sp. n. and C. verrucosa sp. n.) differ from their congeners in the detailed morphology of the proboscis hooks and the atrial organs. The proboscis hooks bear a distal small hook in C. verrucosa sp. n., a feature only shared with C. paradoxa. Two of the four new species of Carcharodorhynchus (C. smilodon sp. n. and C. papillaris sp. n. ) are very similar to C. flavidus in that the toothed belts of the proboscis are not continuous. However, they clearly differ from that species and from each other in the detailed construction of the teeth and copulatory organ. Carcharodorhynchus spiniformis sp. n. and C. nativus sp. n. can also be distinguished from the other species of Carcharodorhynchus by the detailed construction of the proboscis teeth and copulatory organ. The six new species of Schizochilus here described show a spiny cirrus around a central stylet. Two species lack the distal sclerotized cap of the copulatory bulb: S. favus sp. n. and S. bueycabonensis sp. n.. All new species of Schizochilus can be distinguished from each other and their congeners by the detailed construction of the hard parts (cirrus and stylet) of the copulatory organ.

A new rhabdocoel of the genus Syndesmis Silliman, 1881 (Umagillidae) is described from the intestine of the New Zealand sea urchin Evechinus chloroticus (Valenciennes, 1846) Mortensen, 1943a. This new species, Syndesmis kurakaikina n. sp., is morphologically distinct and can easily be recognised by its very long (±1 mm) stylet and its bright-red colour. In addition to providing a formal description, we present some observations on reproduction and life history of this new species. Fecundity is comparable to that of other umagillids and the rate of egg production and development increases with temperature. Hatching in this species is induced by intestinal fluids of its host. Relevant to global warming, we assessed the effect of temperature on survival, fecundity, and development. The tests indicate that Syndesmis kurakaikina n. sp. is tolerant of a wide range of temperatures (11-25 °C) and that its temperature optimum lies between 18.0 and 21.5 °C. Egg viability is, however, significantly compromised at the higher end of this temperature range, with expelled egg capsules often being deformed and showing increasingly lower rates of hatching. Given this, a rise in global temperature might increase the risk of Syndesmis kurakaikina n. sp. infecting new hosts and would possibly facilitate the spread of these endosymbionts.

Silver nanoparticles (AgNPs) belong to the most commercialized nanomaterials, used in both consumer products and medical applications. Despite its omnipresence, in-depth knowledge on the potential toxicity of nanosilver is still lacking, especially for developing organisms. Research on vertebrates is limited due to ethical concerns, and planarians are an ideal invertebrate model to study the effects of AgNPs on stem cells and developing tissues in vivo , as regeneration mimics development by triggering massive stem cell proliferation. Our results revealed a strong interference of AgNPs with tissue- and neuroregeneration which was related to an altered stem cell cycle. The presence of a PVP-coating significantly influenced toxicity outcomes, leading to elevated DNA-damage and decreased stem cell proliferation. Non-coated AgNPs had an inhibiting effect on stem cell and early progeny numbers. Overall, regenerating tissues were more sensitive to AgNP toxicity, and careful handling and appropriate decision making is needed in AgNP applications for healing and developing tissues. We emphasize on the importance of AgNP characterization, as we showed that changes in physicochemical properties influence toxicity.

[This corrects the article DOI: 10.1038/s42003-018-0119-2.].

The first records of Polycystididae from Cuba are provided and discussed. In total nine species have been collected, five of which are new to science, one representing a new genus. Polycydora intermedia gen. n. sp. n. shows intermediate features between Polycystis Kölliker, 1845 and Paulodora Marcus, 1948. The ovaries are kidney shaped, with the oocytes arranged in a row, and lack the hard "nozzles" of Paulodora. A female bursa as in Polycystis is present. In P. intermedia gen. n. sp. n. lacks the strong muscle bulb at the male bursa and the accessory glands type I in the male atrium, which are present in Polycystis. The male atrial organs include a prostate stylet type II connected to a free prostate vesicle type II. The four new species (Phonorhynchoides minor sp. n., Phonorhynchopsis capillaris sp. n., Phonorhynchopsis sublinguatus sp. n., Myobulla armenterosi sp. n.) are distinguished from their congeners by the specific shape and length of the male hard copulatory structures. Phonorhynchoides minor sp. n. has the smallest stylets within the genus, and these stylets are more straight than those of the other species of the genus. In Phonorhynchopsis capillaris sp. n. the prostate stylet type IV is only 20% of the length of the accessory stylet type IV, the lowest relative length within the genus. The accessory stylet in this species is extremely thin; it is only 2 µm wide. Phonorhynchopsis sublinguatus sp. n. has a prostate stylet widened in the distal half, ending in a rounded tip, not twisted in the middle. Because of its strong resemblance to M. armenterosi sp. n., M. berti sp. n., a new species of Myobulla Artois Schockaert, 2000 collected at the Atlantic coast of Panama is described here too. Both species of Myobulla have a prostate stylet type III that shows a 90° turn at some point, as is the case in some other species of Myobulla. In M. berti sp. n. the stylet is smaller overall and is bent in the middle; in M. armenterosi sp. n. the stylet is larger and it is bent more distally. All of the four known species have a very wide geographical distribution: Phonorhynchopsis haegheni, Alcha evelinae, Paraustrorhynchus elixus, and Gyratrix hermaphroditus.

Accurate assessments of biodiversity are crucial to advising ecosystem-monitoring programs and understanding ecosystem function. Nevertheless, a standard operating procedure to assess biodiversity accurately and consistently has not been established. This is especially true for meiofauna, a diverse community (>20 phyla) of small benthic invertebrates that have fundamental ecological roles. Recent studies show that metabarcoding is a cost-effective and time-effective method to estimate meiofauna biodiversity, in contrast to morphological-based taxonomy. Here, we compare biodiversity assessments of a diverse meiofaunal community derived by applying multiple taxonomic methods based on comparative morphology, molecular phylogenetic analysis, DNA barcoding of individual specimens, and metabarcoding of environmental DNA. We show that biodiversity estimates are strongly biased across taxonomic methods and phyla. Such biases affect understanding of community structures and ecological interpretations. This study supports the urgency of improving aspects of environmental high-throughput sequencing and the value of taxonomists in correctly understanding biodiversity estimates., Competing Interests: The authors declare no competing interests.

Aiming to in vivo characterize the responses of pluripotent stem cells and regenerative tissues to carcinogenic stress, we employed the highly regenerative organism Schmidtea mediterranea. Its broad regenerative capacities are attributable to a large pool of pluripotent stem cells, which are considered key players in the lower vulnerability toward chemically induced carcinogenesis observed in regenerative organisms. Schmidtea mediterranea is, therefore, an ideal model to study pluripotent stem cell responses with stem cells residing in their natural environment. Including microenvironmental alterations is important, as the surrounding niche influences the onset of oncogenic events. Both short- (3 days) and long-term (17 days) exposures to the genotoxic carcinogen methyl methanesulfonate (50 µM) were evaluated during homeostasis and animal regeneration, two situations that render altered cellular niches. In both cases, MMS-induced DNA damage was observed, which provoked a decrease in proliferation on the short term. The outcome of DNA damage responses following long-term exposure differed between homeostatic and regenerating animals. During regeneration, DNA repair systems were more easily activated than in animals in homeostasis, where apoptosis was an important outcome. Knockdown experiments confirmed the importance of DNA repair systems during carcinogenic exposure in regenerating animals as knockdown of rad51 induced a stem cell-depleted phenotype, after regeneration was completed.

When complementary resources are required for an optimal life cycle, most animals need to move between different habitats. However, the level of connectivity between resources can vary and, hence, influence individuals' behaviour. We show that landscape composition and configuration affect the connectivity between breeding (heathlands) and foraging habitats (extensively-grazed grasslands) of the European Nightjar (Caprimulgus europaeus), a crepuscular insectivorous bird. On a daily basis, nightjars connect breeding and foraging sites by rapidly crossing unsuitable habitats in order to exploit a higher prey biomass in foraging sites. However, low availability of foraging habitat near breeding sites and clustered landscapes greatly increase foraging distance. Birds occupying these sub-optimal breeding areas compensate for longer travels by increasing foraging duration, and their physiology shows increased stress levels. All findings suggest that landscape heterogeneity can affect population dynamics of nightjars. Therefore, we recommend an integrated management approach for this EU-protected bird species.

Specimens of Syndesmis aethopharynx Westervelt & Kozloff, 1990 (Umagillidae, Rhabdocoela, Platyhelminthes) were collected from the intestine of several specimens of the sea urchin Paracentrotus lividus (Lamarck, 1816) [2], Hansson, 2001 at the Greek coast. This represents the first report of a species of Syndesmis from Greece. Our study has revealed several previously-unreported morphological details and intraspecific variation, which are added to the species description. The position of S. aethopharynx within Umagillidae is confirmed for the first time through molecular data (based on nuclear 18S rDNA), using both Bayesian and maximum likelihood analyses., (Copyright © 2017 Elsevier B.V. All rights reserved.)

One of the major challenges in the development of alternative carcinogenicity assays is the prediction of non-genotoxic carcinogens. The variety of non-genotoxic cancer pathways complicates the search for reliable parameters expressing their carcinogenicity. As non-genotoxic and genotoxic carcinogens have different cancer risks, the objective of this study was to develop a concept for an in vivo test, based on flatworm stem cell dynamics, to detect and classify carcinogenic compounds. Our methodology entails an exposure to carcinogenic compounds during the animal's regeneration process, which revealed differences in proliferative responses between non-genotoxic and genotoxic carcinogens during the initial stages of the regeneration process. A proof of concept was obtained after an extensive study of proliferation dynamics of a genotoxic and a non-genotoxic compound. A pilot validation with a limited set of compounds showed that the proposed concept not only enabled a simple prediction of genotoxic and non-genotoxic carcinogens, but also had the power to discriminate between both. We further optimized this discrimination by combining stem cell proliferation responses with a phenotypic screening and by using specific knockdowns. In the future, more compounds will be tested to further validate and prove this concept.

Stem cells or undifferentiated cells can cope more easily with external stresses. To evaluate the impact of toxic compounds on stem cell dynamics in vivo, in relation to other biological responses, we use the carcinogenic element cadmium and the regenerating model organism Macrostomum lignano. Through both BrdU and anti-histone H3 immunostainings, cadmium-induced effects were investigated at different stages of the stem cell cycle. A 24-h exposure to 100 and 250 μM CdCl2 significantly decreased the number of stem cells (neoblasts) in mitosis, whereas the number of cells in the S phase remained unchanged. After this short-term exposure, the ultrastructure of the neoblasts was minimally affected in contrast to the epidermal tissues. These results were supported by gene expression data: transcripts of cdc2 and pig3 were significantly upregulated during all treatments. Both genes are involved in the cell cycle progression and are transcribed in the gonadal region, where stem cells are highly represented. Based on a substantial increase in gene expression of heat shock proteins (HSP) and their high activity in the gonadal region, we hypothesize that these proteins are key players in the protection of stem cells against external stresses. Apart from the strong HSP induction, other protective processes including cell division, apoptosis and anti-oxidative defence, were also activated. We, therefore, conclude that the protection of stem cells against external stressors may be based on the interplay between stem cell maintenance, i.e. repair and recovery through division, on one hand and apoptosis on the other hand. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1217-1228, 2016., (© 2015 Wiley Periodicals, Inc.)

The importance of nerve-derived signalling for correct regeneration has been the topic of research for more than a hundred years, but we are just beginning to identify the underlying molecular pathways of this process. Within the current review, we attempt to provide an extensive overview of the neural influences during early and late phases of both vertebrate and invertebrate regeneration. In general, denervation impairs limb regeneration, but the presence of nerves is not essential for the regeneration of aneurogenic extremities. This observation led to the "neurotrophic factor(s) hypothesis", which states that certain trophic factors produced by the nerves are necessary for proper regeneration. Possible neuron-derived factors which regulate regeneration as well as the denervation-affected processes are discussed., (Copyright © 2015. Published by Elsevier Inc.)

A delicate balance exists between the process of carcinogenesis and tissue regeneration. A number of malignant tumours are considered the outcome of an impaired or incomplete regeneration process, resulting in persistently dividing cells. Regeneration-competent tissues and animals are able to prevent and counteract growth abnormalities and seem to have a low vulnerability to chemical carcinogenesis. Cancer cell survival depends, among other things, on various redox-related mechanisms, which are targets of currently developed therapies. Disadvantages of these therapies are a lack of specificity and drug resistance. As the majority of these redox-related mechanisms also play an important role in successful and coordinated cell functioning and reproduction, the regeneration process offers a unique parallel context for modern cancer research. This review focuses on the interconnections between regeneration and carcinogenesis and how an understanding of regenerative forces and redox-controlled mechanisms could contribute to the identification of new therapeutic targets to block the growth and survival of cancer cells.

Predatory flatworms belonging to the taxon Kalyptorhynchia are characterized by an anterior muscular proboscis that they use to seize prey. In many cases, the proboscis is armed with hooks, derived either from the extracellular matrix that surrounds the muscles or from intracellular deposits in the epithelium covering the proboscis. Glands associated with the proboscis reportedly are venomous; however, there are few direct tests of this hypothesis. This article reviews the structure and current knowledge of the function of the proboscis in the Kalyptorhynchia, points to areas in which the current understanding of phylogenetic relationships within this taxon is incongruent with our hypothesis of how the proboscis evolved, and addresses areas in need of further research, especially as regards functional morphology and biomechanics., (© The Author 2015. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology. All rights reserved. For permissions please email: journals.permissions@oup.com.)

To investigate hydrophobic test compounds in toxicological studies, solvents like dimethylsulfoxide (DMSO) are inevitable. However, using these solvents, the interpretation of test compound-induced responses can be biased. DMSO concentration guidelines are available, but are mostly based on acute exposures involving one specific toxicity endpoint. Hence, to avoid solvent-toxicant interference, we use multiple chronic test endpoints for additional interpretation of DMSO concentrations and propose a statistical model to assess possible synergistic, antagonistic or additive effects of test compounds and their solvents. In this study, the effects of both short- (1 day) and long-term (2 weeks) exposures to low DMSO concentrations (up to 1000 µl l(-1) ) were studied in the planarian Schmidtea mediterranea. We measured different biological levels in both fully developed and developing animals. In a long-term exposure set-up, a concentration of 500 µl l(-1) DMSO interfered with processes on different biological levels, e.g. behaviour, stem cell proliferation and gene expression profiles. After short exposure times, 500 µl l(-1) DMSO only affected motility, whereas the most significant changes on different parameters were observed at a concentration of 1000 µl l(-1) DMSO. As small sensitivity differences exist between biological levels and developmental stages, we advise the use of this solvent in concentrations below 500 µl l(-1) in this organism. In the second part of our study, we propose a statistical approach to account for solvent-toxicant interactions and discuss full-scale solvent toxicity studies. In conclusion, we reassessed DMSO concentration limits for different experimental endpoints in the planarian S. mediterranea., (Copyright © 2014 John Wiley & Sons, Ltd.)

Recent research highlighted the impact of ROS as upstream regulators of tissue regeneration. We investigated their role and targeted processes during the regeneration of different body structures using the planarian Schmidtea mediterranea, an organism capable of regenerating its entire body, including its brain. The amputation of head and tail compartments induces a ROS burst at the wound site independently of the orientation. Inhibition of ROS production by diphenyleneiodonium (DPI) or apocynin (APO) causes regeneration defaults at both the anterior and posterior wound sites, resulting in reduced regeneration sites (blastemas) and improper tissue homeostasis. ROS signaling is necessary for early differentiation and inhibition of the ROS burst results in defects on the regeneration of the nervous system and on the patterning process. Stem cell proliferation was not affected, as indicated by histone H3-P immunostaining, fluorescence-activated cell sorting (FACS), in situ hybridization of smedwi-1, and transcript levels of proliferation-related genes. We showed for the first time that ROS modulate both anterior and posterior regeneration in a context where regeneration is not limited to certain body structures. Our results indicate that ROS are key players in neuroregeneration through interference with the differentiation and patterning processes.

Some taxonomic groups are less amenable to mitochondrial DNA barcoding than others. Due to the paucity of molecular information of understudied groups and the huge molecular diversity within flatworms, primer design has been hampered. Indeed, all attempts to develop universal flatworm-specific COI markers have failed so far. We demonstrate how high molecular variability and contamination problems limit the possibilities for barcoding using standard COI-based protocols in flatworms. As a consequence, molecular identification methods often rely on other widely applicable markers. In the case of Monogenea, a very diverse group of platyhelminth parasites, and Rhabdocoela, representing one-fourth of all free-living flatworm taxa, this has led to a relatively high availability of nuclear ITS and 18S/28S rDNA sequences on GenBank. In a comparison of the effectiveness in species assignment we conclude that mitochondrial and nuclear ribosomal markers perform equally well. In case intraspecific information is needed, rDNA sequences can guide the selection of the appropriate (i.e. taxon-specific) COI primers if available.