Flupyradifurone negatively affects survival, physical condition and mobility in the two-spotted lady beetle (Adalia bipunctata).

Lady beetles play a crucial role in natural ecosystems and agricultural settings. Unfortunately, these insects and more specifically the two-spotted lady beetle (Adalia bipunctata) are currently facing a severe decline in populations due to various stressors, with pesticide exposure being a significant threat. Flupyradifurone is a relatively newly introduced insecticide and as existing research is mainly elucidating its effects on bees there remains a limited understanding of its effects on non-hymenopteran insects, including lady beetles. In this study we investigated the impact of acute orally applied flupyradifurone doses on survival and sublethal parameters such as physical condition and mobility on A. bipunctata. Our findings revealed a significant increase in mortality among individuals subjected to flupyradifurone doses of 19 ng/individual (corresponding to >1.5-2.0 ng active substance (a.s.)/mg body weight (bw). The calculated LD50 of flupyradifurone at 48 h was 2.11 ng a.s./mg bw corresponding to an amount of 26.38 ng/individual. Sublethal consequences were observable immediately after pesticide application. Even at doses as low as 2 ng/individual (corresponding to >0.0-0.5 ng a.s./mg bw), flupyradifurone induced trembling and temporary immobility in treated animals. Furthermore, pesticide intoxication led to hypoactivity, with less distance covered and a decline in straightness of locomotion. In conclusion, our study underscores the harmful effects of flupyradifurone on the two-spotted lady beetle at doses notably lower than those affecting bees. These findings stress the importance of additional research to attain a more holistic understanding of pesticide impacts not only on a broader range of non-target arthropods species, but also on various exposure routes as well as lethal and sublethal effects.

Influence of the pesticide flupyradifurone on mobility and physical condition of larval green lacewings.

Global pesticide use in agriculture is one reason for the rapid insect decline in recent years. The relatively new pesticide flupyradifurone is neurotoxic to pest insects but considered harmless to bees according to previous risk assessments. With this study, we aim to investigate lethal and sublethal effects of flupyradifurone on larvae of the beneficial arthropod Chrysoperla carnea. We treated the animals orally with field-realistic concentrations of flupyradifurone and examined lethality as well as effects on condition, mobility and locomotion. For the lethal dose 50, we determined a value of > 120-200 ng/mg (corresponding to a mean amount of 219 ng/larva) after 168 h. Abnormal behaviors such as trembling and comatose larvae were observed even at the lowest concentration applied (> 0-20 ng/mg, 59 ng/larva). Mobility analysis showed impaired activity patterns, resulting in acute hypoactivity at all pesticide concentrations and time-delayed hyperactivity in larvae treated with > 40-60 ng/mg (100 ng/larva) and > 80-100 ng/mg (120 ng/larva), respectively. Even locomotion as a fundamental behavioral task was negatively influenced throughout larval development. In conclusion, our results demonstrate that flupyradifurone impacts life and survival of lacewing larvae and may pose-despite its status as bee-friendly-a major threat to insect fauna and environment.

Mechanosensory pathways of scorpion pecten hair sensillae-Adjustment of body height and pecten position.

Scorpions' sensory abilities are intriguing, especially the rather enigmatic ventral comb-like chemo- and mechanosensory organs, the so-called pectines. Attached ventrally to the second mesosomal segment just posterior to the coxae of the fourth walking leg pair, the pectines consist of the lamellae, the fulcra, and a variable number of pecten teeth. The latter contain the bimodal peg sensillae, used for probing the substrate with regard to chemo- and mechanosensory cues simultaneously. In addition, the lamellae, the fulcra and the pecten teeth are equipped with pecten hair sensillae (PHS) to gather mechanosensory information. Previously, we have analyzed the neuronal pathway associated with the peg sensillae unraveling their somatotopic projection pattern in dedicated pecten neuropils. Little is known, however, regarding the projections of PHS within the scorpion nervous system. Behavioral and electrophysiological assays showed involvement of PHS in reflexive responses but how the information is integrated remains unresolved. Here, we unravel the innervation pattern of the mechanosensory pecten hair afferents in Mesobuthus eupeus and Euscorpius italicus. By using immunofluorescent labeling and injection of Neurobiotin tracer, we identify extensive arborizations of afferents, including (i) ventral neuropils, (ii) somatotopically organized multisegmental sensory tracts, (iii) contralateral branches via commissures, and (iv) direct ipsilateral innervation of walking leg neuromeres 3 and 4. Our results suggest that PHS function as sensors to elicit reflexive adjustment of body height and obstacle avoidance, mediating accurate pecten teeth alignment to guarantee functionality of pectines, which are involved in fundamental capacities like mating or navigation.

Locomotion in the pseudoscorpion Chelifer cancroides: forward, backward and upside-down walking in an eight-legged arthropod.

While insect locomotion has been intensively studied, there are comparably few studies investigating octopedal walking behaviour, and very little is known about pseudoscorpions in particular. Therefore, we performed an extensive locomotion analysis during forward, backward and upside-down walking in the cosmopolitan pseudoscorpion Chelifer cancroides. During forward locomotion, we observed C. cancroides to freeze locomotion frequently for short time periods. These microstops were barely visible to the naked eye with a duration of 100-200 ms. Our locomotion analysis revealed that C. cancroides performs a statically stable and highly coordinated alternating tetrapod gait during forward and backward walking, with almost complete inversion of the tetrapod schemes, but no rigidly fixed leg coordination during upside-down walks with low walking speeds up to 4 body lengths per second. Highest speeds (up to 17 body lengths per second), mainly achieved by consistent leg coordination and strong phase shifts, were observed during backward locomotion (escape behaviour), whereas forward walking was characterised by lower speeds and phase shifts of ∼10% between two loosely coupled leg groups within one tetrapod. That is, during the movement of one tetrapod group, the last and the third leg are almost synchronous in their swing phases, as are the second and the first leg. A special role of the second leg pair was demonstrated, probably mainly for stability reasons and related to the large pedipalps.

Anatomy of the Nervous System in Chelifer cancroides (Arachnida: Pseudoscorpiones) with a Distinct Sensory Pathway Associated with the Pedipalps.

Many arachnid taxa have evolved unique, highly specialized sensory structures such as antenniform legs in Amblypygi (whip spiders), for instance, or mesosomal pectines in scorpions. Knowledge of the neuroanatomy as well as functional aspects of these sensory organs is rather scarce, especially in comparison to other arthropod clades. In pseudoscorpions, no special sensory structures have been discovered so far. Nevertheless, these animals possess dominant, multifunctional pedipalps, which are good candidates for being the primary sensory appendages. However, only little is known about the anatomy of the nervous system and the projection pattern of pedipalpal afferents in this taxon. By using immunofluorescent labeling of neuronal structures as well as lipophilic dye labeling of pedipalpal pathways, we identified the arcuate body, as well as a comparatively small mushroom body, the latter showing some similarities to that of Solifugae (sun spiders and camel spiders). Furthermore, afferents from the pedipalps terminate in a glomerular and a layered neuropil. Due to the innervation pattern and structural appearance, we conclude that these neuropils are the first integration centers of the chemosensory and mechanosensory afferents. Within Arthropoda, but also other invertebrates or even vertebrates, sensory structures show rather similar neuronal arrangement. Thus, these similarities in the sensory systems of different evolutionary origin have to be interpreted as functional prerequisites of the respective modality.

Structure of the pecten neuropil pathway and its innervation by bimodal peg afferents in two scorpion species.

The pectines of scorpions are comb-like structures, located ventrally behind the fourth walking legs and consisting of variable numbers of teeth, or pegs, which contain thousands of bimodal peg sensillae. The associated neuropils are situated ventrally in the synganglion, extending between the second and fourth walking leg neuromeres. While the general morphology is consistent among scorpions, taxon-specific differences in pecten and neuropil structure remain elusive but are crucial for a better understanding of chemosensory processing. We analysed two scorpion species (Mesobuthus eupeus and Heterometrus petersii) regarding their pecten neuropil anatomy and the respective peg afferent innervation with anterograde and lipophilic tracing experiments, combined with immunohistochemistry and confocal laser-scanning microscopy. The pecten neuropils consisted of three subcompartments: a posterior pecten neuropil, an anterior pecten neuropil and a hitherto unknown accessory pecten neuropil. These subregions exhibited taxon-specific variations with regard to compartmentalisation and structure. Most notable were structural differences in the anterior pecten neuropils that ranged from ovoid shape and strong fragmentation in Heterometrus petersii to elongated shape with little compartmentalisation in Mesobuthus eupeus. Labelling the afferents of distinct pegs revealed a topographic organisation of the bimodal projections along a medio-lateral axis. At the same time, all subregions along the posterior-anterior axis were innervated by a single peg's afferents. The somatotopic projection pattern of bimodal sensillae appears to be common among arachnids, including scorpions. This includes the structure and organisation of the respective neuropils and the somatotopic projection patterns of chemosensory afferents. Nonetheless, the scorpion pecten pathway exhibits unique features, e.g. glomerular compartmentalisation superimposed on somatotopy, that are assumed to allow high resolution of substrate-borne chemical gradients.

Non-visual homing and the current status of navigation in scorpions.

Within arthropods, the investigation of navigational aspects including homing abilities has mainly focused on insect representatives, while other arthropod taxa have largely been ignored. As such, scorpions are rather underrepresented concerning behavioral studies for reasons such as low participation rates and motivational difficulties. Here, we review the sensory abilities of scorpions related to navigation. Furthermore, we present an improved laboratory setup to shed light on navigational abilities in general and homing behavior in particular. We tracked directed movements towards home shelters of the lesser Asian scorpion Mesobuthus eupeus to give a detailed description of their departure and return movements. To do so, we analyzed the departure and return angles as well as measures of directness like directional deviation, lateral displacement, and straightness indices. We compared these parameters under different light conditions and with blinded scorpions. The motivation of scorpions to leave their shelter depends strongly upon the light condition and the starting time of the experiment; highest participation rates were achieved with infrared conditions or blinded scorpions, and close to dusk. Naïve scorpions are capable of returning to a shelter object in a manner that is directionally consistent with the home vector. The first-occurring homing bouts are characterized by paths consisting of turns about 10 cm to either side of the straightest home path and a distance efficiency of roughly three-quarters of the maximum efficiency. Our results show that neither chemosensation nor vision, but rather path integration based on proprioception, plays a superior role in the homing of scorpions.

Histaminergic interneurons in the ventral nerve cord: assessment of their value for Euarthropod phylogeny.

Despite numerous approaches to the resolution of euarthropod phylogeny, mainly based on modern sequence information and traditional external morphology, the resulting hypotheses are often contradictory and leave many questions about euarthropod evolution unanswered. The comparison of developmental and structural aspects of the nervous system has shown to be a valuable contribution to the assessment of current phylogenetic hypotheses. One promising approach for the generation of new character sets is the morphology of transmitter systems and the discovery of individually identifiable neurons, which allow phylogenetic comparisons on the single cell level. In this context, the serotonin transmitter system has been investigated to a considerable degree. Studies to date have yielded important stimuli to our understanding of euarthropod relationships and the evolution of their nervous systems. However, data on other transmitter systems remain fragmented, and their value with respect to phylogenetic questions remains speculative. The biogenic amine histamine is a promising transmitter; a substantial amount of data has been reported in the literature and the homology of some histaminergic neurons has been suggested. Here, we present a comprehensive review of histaminergic neurons in the ventral nerve cord of Euarthropoda. Using immunocytochemical labeling of histamine combined with confocal laser-scanning microscopy, we investigated the transmitter system in phylogenetically relevant taxa, such as Zygentoma, Remipedia, Diplopoda, and Arachnida. By reconstructing ground patterns, we evaluated the significance of this specific character set for euarthropod phylogeny. With this approach, we identified a set of neurons, which can be considered homologous within the respective major taxon. In conclusion, the histaminergic system contains useful information for our understanding of euarthropod phylogeny, supporting the proposed clades Tetraconata and Mandibulata. Furthermore, this character set has considerable potential to help resolve relationships within the major clades at a deeper level of taxonomy, due to the considerable variability in neurite morphology.

Serotonergic neurons in the ventral nerve cord of Chilopoda - a mandibulate pattern of individually identifiable neurons.

BACKGROUND: Given the numerous hypotheses concerning arthropod phylogeny, independent data are needed to supplement knowledge based on traditional external morphology and modern molecular sequence information. One promising approach involves comparisons of the structure and development of the nervous system. Along these lines, the morphology of serotonin-immunoreactive neurons in the ventral nerve cord has been investigated in numerous tetraconate taxa (Crustacea and Hexapoda). It has been shown that these neurons can be identified individually due to their comparably low number, characteristic soma position, and neurite morphology, thus making it possible to establish homologies at the single cell level. Within Chilopoda (centipedes), detailed analyses of major branching patterns of serotonin-immunoreactive neurons are missing, but are crucial for developing meaningful conclusions on the homology of single cells. RESULTS: In the present study, we re-investigated the distribution and projection patterns of serotonin-immunoreactive neurons in the ventral nerve cord of three centipede species: Scutigera coleoptrata, Lithobius forficatus, and Scolopendra oraniensis. The centipede serotonergic system in the ventral nerve cord contains defined groups of individually identifiable neurons. An anterior and two posterior immunoreactive neurons per hemiganglion with contralateral projections, a pair of ipsilateral projecting lateral neurons (an autapomorphic character for Chilopoda), as well as a postero-lateral group of an unclear number of cells are present in the ground pattern of Chilopoda. CONCLUSIONS: Comparisons to the patterns of serotonin-immunoreactive neurons of tetraconate taxa support the homology of anterior and posterior neurons. Our results thus support a sister group relationship of Myriapoda and Tetraconata and, further, a mandibulate ground pattern of individually identifiable serotonin-immunoreactive neurons in the ventral nerve cord. Medial neurons are not considered to be part of the tetraconate ground pattern, but could favor the 'Miracrustacea hypothesis', uniting Remipedia, Cephalocarida, and Hexapoda.

Serotonin-containing neurons in basal insects: In search of ground patterns among tetraconata.

The ventral nerve cord of Tetraconata contains a comparably low number of serotonin-immunoreactive neurons, facilitating individual identification of cells and their characteristic neurite morphology. This offers the rather unique possibility of establishing homologies at the single cell level. Because phylogenetic relationships within Tetraconata are still discussed controversially, comparisons of individually identifiable neurons can help to unravel these issues. Serotonin immunoreactivity has been investigated in numerous tetraconate taxa, leading to reconstructions of hypothetical ground patterns for major lineages. However, detailed descriptions of basal insects are still missing, but are crucial for meaningful evolutionary considerations. We investigated the morphology of individually identifiable serotonin-immunoreactive neurons in the ventral nerve cord of Zygentoma (Thermobia domestica, Lepisma saccharina, Atelura formicaria) and Archaeognatha (Machilis germanica, Dilta hibernica). To improve immunocytochemical resolution, we also performed preincubation experiments with 5-hydroxy-L-tryptophan and serotonin. Additionally, we checked for immunolabeling of tryptophan hydroxylase, an enzyme associated with the synthesis of serotonin. Besides the generally identified groups of anterolateral, medial, and posterolateral neurons within each ganglion of the ventral nerve cord, we identified several other immunoreactive cells, which seem to have no correspondence in other tetraconates. Furthermore, we show that not all immunoreactive neurons produce serotonin, but have the capability for serotonin uptake. Comparisons with the patterns of serotonin-containing neurons in major tetraconate taxa suggest a close phylogenetic relationship of Remipedia, Cephalocarida, and Hexapoda, supporting the Miracrustacea hypothesis. J. Comp. Neurol., 2016. © 2016 Wiley Periodicals, Inc. J. Comp. Neurol. 525:79-115, 2017. © 2016 Wiley Periodicals, Inc.

Olfactory pathway in Xibalbanus tulumensis: remipedian hemiellipsoid body as homologue of hexapod mushroom body.

The Remipedia have been proposed to be the crustacean sister group of the Hexapoda. These blind cave animals heavily rely on their chemical sense and are thus rewarding subjects for the analysis of olfactory pathways. The evolution of these pathways as a character for arthropod phylogeny has recently received increasing attention. Here, we investigate the situation in Xibalbanus tulumensis by focal dye injections and immunolabelling of the catalytic subunit of the cAMP-dependent protein kinase (DC0), an enzyme particularly enriched in insect mushroom bodies. DC0 labelling of the hemiellipsoid body suggests its subdivision into a cap-like and a core neuropil. Immunofluorescence of the enzyme glutamic acid decarboxylase (GAD), which synthesizes γ-aminobutyric acid (GABA), has revealed a cluster of GABAergic interneurons in the hemiellipsoid body, reminiscent of the characteristic feedback neurons of the mushroom body. Thus, the hemiellipsoid body of Xibalbanus shares many of the characteristics of insect mushroom bodies. Nevertheless, the general neuroanatomy of the olfactory pathway in the Remipedia strongly corresponds to the malacostracan ground pattern. Given that the Remipedia are probably the sister group of the Hexapoda, the phylogenetic appearance of the typical neuropilar compartments in the insect mushroom body has to be assigned to the origins of the Hexapoda.

The brain in three crustaceans from cavernous darkness.

BACKGROUND: While a number of neuroanatomical studies in other malacostracan taxa have recently contributed to the reconstruction of the malacostracan ground pattern, little is known about the nervous system in the three enigmatic blind groups of peracarids from relict habitats, Thermosbaenacea, Spelaeogriphacea, and Mictocarididae. This first detailed description of the brain in a representative of each taxon is largely based on a combination of serial semi-thin sectioning and computer-aided 3D-reconstructions. In addition, the mictocaridid Mictocaris halope was studied with a combination of immunolabeling (tubulin, nuclear counter-stains) and confocal laser scanning microscopy, addressing also the ventral nerve cord. RESULTS: Adjacent to the terminal medulla, all three representatives exhibit a distal protocerebral neuropil, which is reminiscent of the lobula in other Malacostraca, but also allows for an alternative interpretation in M. halope and the thermosbaenacean Tethysbaena argentarii. A central complex occurs in all three taxa, most distinctively in the spelaeogriphacean Spelaeogriphus lepidops. The deutocerebral olfactory lobe in M. halope and S. lepidops is large. The comparably smaller olfactory lobe in T. argentarii appears to be associated with a unique additional deutocerebral neuropil. A small hemiellipsoid body exists only in the protocerebrum of T. argentarii. Distinctive mechanosensory neuropils corresponding to other malacostracans are missing. CONCLUSIONS: The considerable reduction of the optic lobe in the studied taxa is higher than in any other blind malacostracan. The large size of deutocerebral olfactory centers implies an important role of the olfactory sense. The presence of a distinctive central complex in the blind S. lepidops adds further support to a central-coordinating over a visual function of this structure. The lack of a hemiellipsoid body in M. halope and S. lepidops suggests that their terminal medulla takes over the function of a second order olfactory center completely, as in some other peracarids. The reduction of the optic lobe and hemiellipsoid body is suggested to have occurred several times independently within Peracarida. The missing optic sense in the studied taxa is not correlated with an emphasized mechanosense.

Olfactory projection neuron pathways in two species of marine Isopoda (Peracarida, Malacostraca, Crustacea).

The neuroanatomy of the olfactory pathway has been intensely studied in many representatives of Malacostraca. Nevertheless, the knowledge about bilateral olfactory integration pathways is mainly based on Decapoda. Here, we investigated the olfactory projection neuron pathway of two marine isopod species, Saduria entomon and Idotea emarginata, by lipophilic dye injections into the olfactory neuropil. We show that both arms of the olfactory globular tract form a chiasm in the center of the brain, as known from several other crustaceans. Furthermore, the olfactory projection neurons innervate both the medulla terminalis and the hemiellipsoid body of the ipsi- and the contralateral hemisphere. Both protocerebral neuropils are innervated to a comparable extent. This is reminiscent of the situation in the basal decapod taxon Dendrobranchiata. Thus, we propose that an innervation by the olfactory globular tract of both the medulla terminalis and the hemiellipsoid body is characteristic of the decapod ground pattern, but also of the ground pattern of Caridoida.

Serotonin-immunoreactive neurons in the ventral nerve cord of Remipedia (Crustacea): support for a sister group relationship of Remipedia and Hexapoda?

BACKGROUND: Remipedia were initially seen as a primitive taxon within Pancrustacea based on characters considered ancestral, such as the homonomously segmented trunk. Meanwhile, several morphological and molecular studies proposed a more derived position of Remipedia within Pancrustacea, including a sister group relationship to Hexapoda. Because of these conflicting hypotheses, fresh data are crucial to contribute new insights into euarthropod phylogeny. The architecture of individually identifiable serotonin-immunoreactive neurons has successfully been used for phylogenetic considerations in Euarthropoda. Here, we identified neurons in three species of Remipedia with an antiserum against serotonin and compared our findings to reconstructed ground patterns in other euarthropod taxa. Additionally, we traced neurite connectivity and neuropil outlines using antisera against acetylated α-tubulin and synapsin. RESULTS: The ventral nerve cord of Remipedia displays a typical rope-ladder-like arrangement of separate metameric ganglia linked by paired longitudinally projecting connectives. The peripheral projections comprise an intersegmental nerve, consisting of two branches that fuse shortly after exiting the connectives, and the segmental anterior and posterior nerve. The distribution and morphology of serotonin-immunoreactive interneurons in the trunk segments is highly conserved within the remipede species we analyzed, which allows for the reconstruction of a ground pattern: two posterior and one anterior pair of serotonin-immunoreactive neurons that possess a single contralateral projection. Additionally, three pairs of immunoreactive neurons are found in the medial part of each hemiganglion. In one species (Cryptocorynetes haptodiscus), the anterior pair of immunoreactive neurons is missing. CONCLUSIONS: The anatomy of the remipede ventral nerve cord with its separate metameric ganglia mirrors the external morphology of the animal's trunk. The rope-ladder-like structure and principal architecture of the segmental ganglia in Remipedia corresponds closely to that of other Euarthropoda. A comparison of the serotonin-immunoreactive cell arrangement of Remipedia to reconstructed ground patterns of major euarthropod taxa supports a homology of the anterior and posterior neurons in Pancrustacea. These neurons in Remipedia possess unbranched projections across the midline, pointing towards similarities to the hexapod pattern. Our findings are in line with a growing number of phylogenetic investigations proposing Remipedia to be a rather derived crustacean lineage that perhaps has close affinities to Hexapoda.

Serotonin immunoreactive interneurons in the brain of the Remipedia: new insights into the phylogenetic affinities of an enigmatic crustacean taxon.

BACKGROUND: Remipedia, a group of homonomously segmented, cave-dwelling, eyeless arthropods have been regarded as basal crustaceans in most early morphological and taxonomic studies. However, molecular sequence information together with the discovery of a highly differentiated brain led to a reconsideration of their phylogenetic position. Various conflicting hypotheses have been proposed including the claim for a basal position of Remipedia up to a close relationship with Malacostraca or Hexapoda. To provide new morphological characters that may allow phylogenetic insights, we have analyzed the architecture of the remipede brain in more detail using immunocytochemistry (serotonin, acetylated α-tubulin, synapsin) combined with confocal laser-scanning microscopy and image reconstruction techniques. This approach allows for a comprehensive neuroanatomical comparison with other crustacean and hexapod taxa. RESULTS: The dominant structures of the brain are the deutocerebral olfactory neuropils, which are linked by the olfactory globular tracts to the protocerebral hemiellipsoid bodies. The olfactory globular tracts form a characteristic chiasm in the center of the brain. In Speleonectes tulumensis, each brain hemisphere contains about 120 serotonin immunoreactive neurons, which are distributed in distinct cell groups supplying fine, profusely branching neurites to 16 neuropilar domains. The olfactory neuropil comprises more than 300 spherical olfactory glomeruli arranged in sublobes. Eight serotonin immunoreactive neurons homogeneously innervate the olfactory glomeruli. In the protocerebrum, serotonin immunoreactivity revealed several structures, which, based on their position and connectivity resemble a central complex comprising a central body, a protocerebral bridge, W-, X-, Y-, Z-tracts, and lateral accessory lobes. CONCLUSIONS: The brain of Remipedia shows several plesiomorphic features shared with other Mandibulata, such as deutocerebral olfactory neuropils with a glomerular organization, innervations by serotonin immunoreactive interneurons, and connections to protocerebral neuropils. Also, we provided tentative evidence for W-, X-, Y-, Z-tracts in the remipedian central complex like in the brain of Malacostraca, and Hexapoda. Furthermore, Remipedia display several synapomorphies with Malacostraca supporting a sister group relationship between both taxa. These homologies include a chiasm of the olfactory globular tract, which connects the olfactory neuropils with the lateral protocerebrum and the presence of hemiellipsoid bodies. Even though a growing number of molecular investigations unites Remipedia and Cephalocarida, our neuroanatomical comparison does not provide support for such a sister group relationship.

Global biodiversity and phylogenetic evaluation of remipedia (crustacea).

Remipedia is one of the most recently discovered classes of crustaceans, first described in 1981 from anchialine caves in the Bahamas Archipelago. The class is divided into the order Enantiopoda, represented by two fossil species, and Nectiopoda, which contains all known extant remipedes. Since their discovery, the number of nectiopodan species has increased to 24, half of which were described during the last decade. Nectiopoda exhibit a disjunct global distribution pattern, with the highest abundance and diversity in the Caribbean region, and isolated species in the Canary Islands and in Western Australia. Our review of Remipedia provides an overview of their ecological characteristics, including a detailed list of all anchialine marine caves, from which species have been recorded. We discuss alternative hypotheses of the phylogenetic position of Remipedia within Arthropoda, and present first results of an ongoing molecular-phylogenetic analysis that do not support the monophyly of several nectiopodan taxa. We believe that a taxonomic revision of Remipedia is absolutely essential, and that a comprehensive revision should include a reappraisal of the fossil record.

Arthropod phylogeny revisited, with a focus on crustacean relationships.

Higher-level arthropod phylogenetics is an intensely active field of research, not least as a result of the hegemony of molecular data. However, not all areas of arthropod phylogenetics have so far received equal attention. The application of molecular data to infer a comprehensive phylogeny of Crustacea is still in its infancy, and several emerging results are conspicuously at odds with morphology-based studies. In this study, we present a series of molecular phylogenetic analyses of 88 arthropods, including 57 crustaceans, representing all the major lineages, with Onychophora and Tardigrada as outgroups. Our analyses are based on published and new sequences for two mitochondrial markers, 16S rDNA and cytochrome c oxidase subunit I (COI), and the nuclear ribosomal gene 18S rDNA. We designed our phylogenetic analyses to assess the effects of different strategies of sequence alignment, alignment masking, nucleotide coding, and model settings. Our comparisons show that alignment optimization of ribosomal markers based on secondary structure information can have a radical impact on phylogenetic reconstruction. Trees based on optimized alignments recover monophyletic Arthropoda (excluding Onychophora), Pancrustacea, Malacostraca, Insecta, Myriapoda and Chelicerata, while Maxillopoda and Hexapoda emerge as paraphyletic groups. Our results are unable to resolve the highest-level relationships within Arthropoda, and none of our trees supports the monophyly of Myriochelata or Mandibulata. We discuss our results in the context of both the methodological variations between different analyses, and of recently proposed phylogenetic hypotheses. This article offers a preliminary attempt to incorporate the large diversity of crustaceans into a single molecular phylogenetic analysis, assessing the robustness of phylogenetic relationships under varying analysis parameters. It throws into sharp relief the relative strengths and shortcomings of the combined molecular data for assessing this challenging phylogenetic problem, and thereby provides useful pointers for future studies.