High-resolution vision in pelagic polychaetes.

High-resolution object vision - the ability to separate, classify, and interact with specific objects in the environment against the visual background - has only been conclusively shown to have evolved in three of the thirty-five animal phyla: chordates, arthropods, and mollusks (cephalopods)1. However, alciopid polychaetes (Phyllodocidae, Alciopini), which possess a pair of bulbous camera-type eyes, have also been hypothesized to achieve high acuity. In this study, we examined three species of night-active pelagic alciopids from the Mediterranean Sea. Our optical, morphological, and electrophysiological investigations show that their eyes have high spatial acuity and temporal resolution, supporting the notion that they are capable of active, high-resolution object vision. These results encourage interesting hypotheses about the visual ecology of these enigmatic polychaetes.

Associative learning in the box jellyfish Tripedalia cystophora.

Associative learning, such as classical or operant conditioning, has never been unequivocally associated with animals outside bilatarians, e.g., vertebrates, arthropods, or mollusks. Learning modulates behavior and is imperative for survival in the vast majority of animals. Obstacle avoidance is one of several visually guided behaviors in the box jellyfish, Tripedalia cystophora Conant, 1897 (Cnidaria: Cubozoa), and it is intimately associated with foraging between prop roots in their mangrove habitat. The obstacle avoidance behavior (OAB) is a species-specific defense reaction (SSDR) for T. cystophora, so identifying such SSDR is essential for testing the learning capacity of a given animal. Using the OAB, we show that box jellyfish performed associative learning (operant conditioning). We found that the rhopalial nervous system is the learning center and that T. cystophora combines visual and mechanical stimuli during operant conditioning. Since T. cystophora has a dispersed central nervous system lacking a conventional centralized brain, our work challenges the notion that associative learning requires complex neuronal circuitry. Moreover, since Cnidaria is the sister group to Bilateria, it suggests the intriguing possibility that advanced neuronal processes, like operant conditioning, are a fundamental property of all nervous systems.

Unique horizontal gaze control in the box jellyfish, Tripedalia cystophora.

All known cubozoans, box jellyfish, have a similar visual system. They possess four sensory structures called rhopalia, which carry-six eyes each. Two of these six eyes are true image-forming camera type eyes in several ways similar to vertebrate eyes. The rhopalia hang by a thin flexible stalk and in the distal end, there is a high-density crystal. In an earlier study of the Caribbean species Tripedalia cystophora, we showed that the crystals act as weights ensuring that the rhopalia are always upright no matter the orientation of the medusa and the vertical part of the visual field of the eyes thus kept relatively constant. Here we have examined the horizontal part of the visual field under different experimental conditions including different visual environments. We find that the horizontal gaze direction is largely controlled by the anatomy of the rhopalium and rhopalial stalk, similar to what has previously been shown for the vertical gaze direction. In a vertically oriented medusa, the rhopalia are kept with a 90° angle between them with the lower lens eyes (LLE) pointing inwards. This 90° shift is kept in horizontally swimming medusa, resulting in the left LLE gazing right, the right gazing left, the bottom gazing orally (backwards compared to swimming direction), and the top LLE gazing aborally (forwards compared to swimming direction). The light environment was manipulated to test if the visual input influences this seemingly strict horizontal gaze direction but even in complete darkness there is tight mechanistic control.

The contraction-expansion behaviour in the demosponge Tethya wilhelma is light controlled and follows a diurnal rhythm.

Sponges (phylum Porifera) are metazoans which lack muscles and nerve cells, yet perform coordinated behaviours such as whole-body contractions. Previous studies indicate diurnal variability in both the number of contractions and the expression of circadian clock genes. Here, we show that diurnal patterns are present in the contraction-expansion behaviour of the demosponge Tethya wilhelma, by using infrared videography and a simulated night/day cycle including sunrise and sunset mimics. In addition, we show that this behaviour is at least strongly influenced by ambient light intensity and therefore indicates light-sensing capabilities in this sponge species. This is supported by our finding that T. wilhelma consistently contracts at sunrise, and that this pattern disappears both when the sponge is kept in constant darkness and when it is in constant light.

Corrigendum: Expression of opsins of the box jellyfish Tripedalia cystophora reveals the first photopigment in cnidarian ocelli and supports the presence of photoisomerases.

[This corrects the article DOI: 10.3389/fnana.2022.916510.].

Expression of Opsins of the Box Jellyfish Tripedalia cystophora Reveals the First Photopigment in Cnidarian Ocelli and Supports the Presence of Photoisomerases.

Cubomedusae, or box jellyfish, have a complex visual system comprising 24 eyes of four types. Like other cnidarians, their photoreceptor cells are ciliary in morphology, and a range of different techniques together show that at least two of the eye types-the image-forming upper and lower lens eyes-express opsin as the photopigment. The photoreceptors of these two eye types express the same opsin (Tc LEO), which belongs to the cnidarian-specific clade cnidops. Interestingly, molecular work has found a high number of opsin genes in box jellyfish, especially in the Caribbean species Tripedalia cystophora, most of which are of unknown function. In the current study, we raised antibodies against three out of five opsins identified from transcriptomic data from T. cystophora and used them to map the expression patterns. These expression patterns suggest one opsin as the photopigment in the slit eyes and another as a putative photoisomerase found in photoreceptors of all four eyes types. The last antibody stained nerve-like cells in the tentacles, in connection with nematocytes, and the radial nerve, in connection with the gonads. This is the first time photopigment expression has been localized to the outer segments of the photoreceptors in a cnidarian ocellus (simple eye). The potential presence of a photoisomerase could be another interesting convergence between box jellyfish and vertebrate photoreceptors, but it awaits final experimental proof.

Have the eyes of bioluminescent scale worms adapted to see their own light? A comparative study of eyes and vision in Harmothoe imbricata and Lepidonotus squamatus.

Annelids constitute a diverse phylum with more than 19,000 species, which exhibit greatly varying morphologies and lifestyles ranging from sessile detritivores to fast swimming active predators. The lifestyle of an animal is closely linked to its sensory systems, not least the visual equipment. Interestingly, many errantian annelid species from different families, such as the scale worms (Polynoidae), have two pairs of eyes on their prostomium. These eyes are typically 100-200 µm in diameter and structurally similar judged from their gross morphology. The polynoids Harmothoe imbricata and Lepidonotus squamatus from the North Atlantic are both benthic predators preying on small invertebrates but only H. imbricata can produce bioluminescence in its scales. Here, we examined the eye morphology, photoreceptor physiology and light-guided behaviour in these two scale worms to assess their visual capacity and visual ecology. The structure and physiology of the two pairs of eyes are remarkably similar within each species, with the only difference being the gaze direction. The photoreceptor physiology, however, differs between species. Both species express a single opsin in their eyes, but in H. imbricata the peak sensitivity is green shifted and the temporal resolution is lower, suggesting that the eyes of H. imbricata are adapted to detect their own bioluminescence. The behavioural experiments showed that both species are strictly night active but yielded no support for the hypothesis that H. imbricata is repelled by its own bioluminescence.

Neuropeptide expression in the box jellyfish Tripedalia cystophora-New insights into the complexity of a "simple" nervous system.

Box jellyfish have an elaborate visual system and perform advanced visually guided behaviors. However, the rhopalial nervous system (RNS), believed to be the main visual processing center, only has 1000 neurons in each of the four eye carrying rhopalia. We have examined the detailed structure of the RNS of the box jellyfish Tripedalia cystophora, using immunolabeling with antibodies raised against four putative neuropeptides (T. cystophora RFamide, VWamide, RAamide, and FRamide). In the RNS, T. cystophora RF-, VW-, and RAamide antibodies stain sensory neurons, the pit eyes, the neuropil, and peptide-specific subpopulations of stalk-associated neurons and giant neurons. Furthermore, RFamide ir+ neurites are seen in the epidermal stalk nerve, whereas VWamide antibodies stain the gastrodermal stalk nerve. RFamide has the most widespread expression including in the ring and radial nerves, the pedalium nerve plexus, and the tentacular nerve net. RAamide is the putative neurotransmitter in the motor neurons of the subumbrellar nerve net, and VWamide is a potential marker for neuronal differentiation as it is found in subpopulations of undifferentiated cells both in the rhopalia and in the bell. The results from the FRamide antibodies were not included as only few cells were stained, and in an unreproducible way. Our studies show hitherto-unseen details of the nervous system of T. cystophora and allowed us to identify specific functional groups of neurons. This identification is important for understanding visual processing in the RNS and enables experimental work, directly addressing the role of the different neuropeptides in vision.

Photoresponses in the radiolar eyes of the fan worm Acromegalomma vesiculosum.

Fan worms (Annelida: Sabellidae) possess compound eyes and other photoreceptors on their radiolar feeding tentacles. These eyes putatively serve as an alarm system that alerts the worm to encroaching threats, eliciting a rapid defensive retraction into their protective tube. The structure and independent evolutionary derivation of these radiolar eyes make them a fascinating target for exploring the emergence of new sensory systems and visually guided behaviours. However, little is known about their physiology and how this impacts their function. Here, we present electroretinogram recordings from the radiolar eyes of the fan worm Acromegalomma vesiculosum We examine their spectral sensitivity along with their dynamic range and temporal resolution. Our results show that they possess one class of photoreceptors with a single visual pigment peaking in the blue-green part of the spectrum around 510 nm, which matches the dominant wavelengths in their shallow coastal habitats. We found the eyes to have a rather high temporal resolution with a critical flicker fusion frequency around 35 Hz. The high temporal resolution of this response is ideally suited for detecting rapidly moving predators but also necessitates downstream signal processing to filter out caustic wave flicker. This study provides a fundamental understanding of how these eyes function. Furthermore, these findings emphasise a set of dynamic physiological principles that are well suited for governing a multi-eyed startle response in coastal aquatic habitats.

Gonadal cnidocytes in the cubozoan Tripedalia cystophora Conant, 1897 (Cnidaria: Cubozoa).

Cubozoans have a complex lifecycle in many ways similar to the scyphozoan lifecycle. The sexual reproduction within cubozoans varies between species with one clade having copulation and internal fertilization and the release of planula larvae. This cubozoan clade, the family Tripedaliidae, includes three species, Copula sivickisi, Tripedalia cystophora, and Tripedalia binata. In a recent study, it was suggested that in C. sivickisi cnidocytes play a new and important role during the sexual reproduction. Male derived cnidocytes anchor sperm packages to the female gonads and female derived cnidocytes protect the externalized embryo strand. Here, we have examined the gonads and gametes of T. cystophora and our results reveal that the male produced spermatozeugmata have a high number of isorhiza type cnidocytes, which are transferred along with the sperm during copulation. This adds further support to our hypothesis that they are important for sperm anchorage. The female gonads are lacking cnidocytes all together showing that cnidocyte production is not just a default state of the epithelium in these animals.

Hyperparasitism in caves: Bats, bat flies and ectoparasitic fungus interaction.

Bat flies (Diptera: Nycteribiinae) are highly specialized bloodsucking bat ectoparasites. Some of the ectoparasitic bat flies are themselves parasitized with an ectoparasitic fungus of the genus Arthrorhynchus (Laboulbeniales). Ascospores of the fungus attach to the cuticle of a bat fly and develop a haustorium that penetrates the host cuticle. This interaction defines the fungus as a hyperparasite. Both the fly and the fungus are obligate parasites and this peculiar case of hyperparasitism has remained largely unstudied. We studied the prevalence of Laboulbeniales, genus Arthrorhynchus, in natural populations of bat flies infesting the bat species Miniopterus schreibersii, Myotis bechsteinii, My. blythii, My. daubentonii, My. escalerai and My. myotis in Portuguese caves. Laboulbeniales were found infecting 10 of the 428 screened bat flies (2.3%) in natural populations, with fewer infections in winter. Images obtained with transmission electron microscopy show the fungal haustorium within the bat fly host tissue, from where it extracts nutrition.

Sources, distribution and fate of microfibres on the Great Barrier Reef, Australia.

Marine microdebris, in particular microplastics (plastics <5 mm), has become an issue of international concern due to its prevalence, persistence and potential adverse impacts on marine ecosystems. Informing source reduction based on ecological effects requires an understanding of the origin, distribution and characteristics of microdebris and the interactions with marine organisms. Here we show widespread contamination of the central Great Barrier Reef environment with microdebris, with microfibres comprising 86% of all items detected. Microdebris intake by coral reef fish was non-random, with chemical composition, shape and colour differing significantly from that detected in surface waters. Furthermore, the origin of microdebris contamination in surface waters is non-random with riverine discharge a likely source for microdebris detected at inshore, but not at offshore reef locations. Our findings demonstrate the complexities associated with determining marine microdebris exposure and fate, and assist in improving future ecological assessments and prioritizing source reduction.

Eyes and negative phototaxis in juvenile crown-of-thorns starfish, Acanthaster species complex.

As a corallivore, the crown-of-thorns starfish (COTS; Acanthaster species complex), has significant impacts on coral mortality and community structure on tropical reefs throughout its Indo-Pacific range. COTS form aggregations which systematically move through and across reefs causing significant loss in hard coral cover. Previous work has shown that their behaviours on the reef are influenced by rheotaxis, olfaction and vision, with vision guiding adult animals to their coral habitat at short distances. As the compound eye of starfish grows throughout life the visual capacity of juvenile eyes is putatively less than for adult animals. Here we show this to be the case. Juvenile eyes have approximately the same visual field as adult eyes but significantly lower spatial resolution. They display negative phototaxis, as observed in adults, but we found no direct proof for the use of spatial resolution in this behaviour. Our results show that juveniles are able to use their eyes to locate their habitat: the coral reef. However, their putatively lower spatial resolution would make this visual task more difficult than for the adults.This article has an associated First Person interview with the first author of the paper.

De novo transcriptome assembly of the cubomedusa Tripedalia cystophora, including the analysis of a set of genes involved in peptidergic neurotransmission.

BACKGROUND: The phyla Cnidaria, Placozoa, Ctenophora, and Porifera emerged before the split of proto- and deuterostome animals, about 600 million years ago. These early metazoans are interesting, because they can give us important information on the evolution of various tissues and organs, such as eyes and the nervous system. Generally, cnidarians have simple nervous systems, which use neuropeptides for their neurotransmission, but some cnidarian medusae belonging to the class Cubozoa (box jellyfishes) have advanced image-forming eyes, probably associated with a complex innervation. Here, we describe a new transcriptome database from the cubomedusa Tripedalia cystophora. RESULTS: Based on the combined use of the Illumina and PacBio sequencing technologies, we produced a highly contiguous transcriptome database from T. cystophora. We then developed a software program to discover neuropeptide preprohormones in this database. This script enabled us to annotate seven novel T. cystophora neuropeptide preprohormone cDNAs: One coding for 19 copies of a peptide with the structure pQWLRGRFamide; one coding for six copies of a different RFamide peptide; one coding for six copies of pQPPGVWamide; one coding for eight different neuropeptide copies with the C-terminal LWamide sequence; one coding for thirteen copies of a peptide with the RPRAamide C-terminus; one coding for four copies of a peptide with the C-terminal GRYamide sequence; and one coding for seven copies of a cyclic peptide, of which the most frequent one has the sequence CTGQMCWFRamide. We could also identify orthologs of these seven preprohormones in the cubozoans Alatina alata, Carybdea xaymacana, Chironex fleckeri, and Chiropsalmus quadrumanus. Furthermore, using TBLASTN screening, we could annotate four bursicon-like glycoprotein hormone subunits, five opsins, and 52 other family-A G protein-coupled receptors (GPCRs), which also included two leucine-rich repeats containing G protein-coupled receptors (LGRs) in T. cystophora. The two LGRs are potential receptors for the glycoprotein hormones, while the other GPCRs are candidate receptors for the above-mentioned neuropeptides. CONCLUSIONS: By combining Illumina and PacBio sequencing technologies, we have produced a new high-quality de novo transcriptome assembly from T. cystophora that should be a valuable resource for identifying the neuronal components that are involved in vision and other behaviors in cubomedusae.

The crowns have eyes: multiple opsins found in the eyes of the crown-of-thorns starfish Acanthaster planci.

BACKGROUND: Opsins are G protein-coupled receptors used for both visual and non-visual photoreception, and these proteins evolutionarily date back to the base of the bilaterians. In the current sequencing age, phylogenomic analysis has proven to be a powerful tool, facilitating the increase in knowledge about diversity within the opsin subclasses and, so far, at least nine types of opsins have been identified. Within echinoderms, opsins have been studied in Echinoidea and Ophiuroidea, which do not possess proper image forming eyes, but rather widely dispersed dermal photoreceptors. However, most species of Asteroidea, the starfish, possess true eyes and studying them will shed light on the diversity of opsin usage within echinoderms and help resolve the evolutionary history of opsins. RESULTS: Using high-throughput RNA sequencing, we have sequenced and analyzed the transcriptomes of different Acanthaster planci tissue samples: eyes, radial nerve, tube feet and a mixture of tissues from other organs. At least ten opsins were identified, and eight of them were found significantly differentially expressed in both eyes and radial nerve, with R-opsin being the most highly expressed in the eye. CONCLUSION: This study provides new important insight into the involvement of opsins in visual and nonvisual photoreception. Of relevance, we found the first indication of an r-opsin photopigment expressed in a well-developed visual eye in a deuterostome animal. Additionally, we provided tissue specific A. planci transcriptomes that will aid in future Evo Devo studies.

Vision Made Easy: Cubozoans Can Advance Our Understanding of Systems-Level Visual Information Processing.

Animals relying on vision as their main sensory modality reserve a large part of their central nervous system to appropriately navigate their environment. In general, neural involvement correlates to the complexity of the visual system and behavioural repertoire. In humans, one third of the available neural capacity supports our single-chambered general-purpose eyes, whereas animals with less elaborate visual systems need less computational power, and generally have smaller brains, and thereby lack in visual behaviour. As a consequence, both traditional model animals (mice, zebrafish, and flies) and more experimentally tractable animals (Hydra, Planaria, and C. elegans) cannot contribute to our understanding of systems-level visual information processing-a Goldilocks case of too big and too small.However, one animal, the box jellyfish Tripedalia cystophora, possesses a rather complex visual system, displays multiple visual behaviours, yet processes visual information by means of a relatively simple central nervous system. This-just right-model system could not only provide information on how visual stimuli are processed through distinct combinations of neural circuitry but also provide a processing algorithm for extracting specific information from a complex visual scene.

Regeneration of the Rhopalium and the Rhopalial Nervous System in the Box Jellyfish Tripedalia cystophora.

Cubozoans have the most intricate visual apparatus within Cnidaria. It comprises four identical sensory structures, the rhopalia, each of which holds six eyes of four morphological types. Two of these eyes are camera-type eyes that are, in many ways, similar to the vertebrate eye. The visual input is used to control complex behaviors, such as navigation and obstacle avoidance, and is processed by an elaborate rhopalial nervous system. Several studies have examined the rhopalial nervous system, which, despite a radial symmetric body plan, is bilaterally symmetrical, connecting the two sides of the rhopalium through commissures in an extensive neuropil. The four rhopalia are interconnected by a nerve ring situated in the oral margin of the bell, and together these structures constitute the cubozoan central nervous system. Cnidarians have excellent regenerative capabilities, enabling most species to regenerate large body areas or body parts, and some species can regenerate completely from just a few hundred cells. Here we test whether cubozoans are capable of regenerating the rhopalia, despite the complexity of the visual system and the rhopalial nervous system. The results show that the rhopalia are readily regrown after amputation and have developed most, if not all, neural elements within two weeks. Using electrophysiology, we investigated the functionality of the regrown rhopalia and found that they generated pacemaker signals and that the lens eyes showed a normal response to light. Our findings substantiate the amazing regenerative ability in Cnidaria by showing here the complex sensory system of Cubozoa, a model system proving to be highly applicable in studies of neurogenesis.

Deep-sea starfish from the Arctic have well-developed eyes in the dark.

Asteroids, starfish, are important members of the macro-benthos in almost all marine environments including the deep sea. Starfish are in general assumed to be largely olfactory guided, but recent studies have shown that two tropical shallow water species rely on vision alone to find their habitat at short distances. Their compound eyes are found at the tip of each arm and they vary little between examined species. Still, nothing is known about vision in the species found in the aphotic zone of the deep sea or whether they even have eyes. Here, 13 species of starfish from Greenland waters, covering a depth range from shallow waters to the deep sea below 1000 m, were examined for the presence of eyes and optical and morphological examinations were used to estimate the quality of vision. Further, species found in the aphotic zone below 320 m were checked for bioluminescence. All species, except the infaunal Ctenodiscus crispatus, had eyes, and two were found to be bioluminescent. Interestingly, one of the species found in the aphotic zone, Novodinia americana, had close to the highest spatial resolution known for starfish eyes along with being bioluminescent. Accordingly, we hypothesize that this species communicates visually using bioluminescent flashes putatively for reproductive purposes. Other species have greatly enhanced sensitivity with few large ommatidia but at the sacrifice of spatial resolution. The discovery of eyes in deep-sea starfish with a huge variation in optical quality and sensitivity indicates that their visual ecology also differs greatly.

No increase in marine microplastic concentration over the last three decades - A case study from the Baltic Sea.

Microplastic is considered a potential threat to marine life as it is ingested by a wide variety of species. Most studies on microplastic ingestion are short-term investigations and little is currently known about how this potential threat has developed over the last decades where global plastic production has increased exponentially. Here we present the first long-term study on microplastic in the marine environment, covering three decades from 1987 to 2015, based on a unique sample set originally collected and conserved for food web studies. We investigated the microplastic concentration in plankton samples and in digestive tracts of two economically and ecologically important planktivorous forage fish species, Atlantic herring (Clupea harengus) and European sprat (Sprattus sprattus), in the Baltic Sea, an ecosystem which is under high anthropogenic pressure and has undergone considerable changes over the past decades. Surprisingly, neither the concentration of microplastic in the plankton samples nor in the digestive tracts changed significantly over the investigated time period. Average microplastic concentration in the plankton samples was 0.21±0.15particlesm-3. Of 814 fish examined, 20% contained plastic particles, of which 95% were characterized as microplastic (<5mm) and of these 93% were fibres. There were no significant differences in the plastic content between species, locations, or time of day the fish were caught. However, fish size and microplastic in the digestive tracts were positively correlated, and the fish contained more plastic during summer than during spring, which may be explained by increased food uptake with size and seasonal differences in feeding activity. This study highlights that even though microplastic has been present in the Baltic environment and the digestive tracts of fishes for decades, the levels have not changed in this period. This underscores the need for greater understanding of how plastic is cycled through marine ecosystems. The stability of plastic concentration and contamination over time observed here indicates that the type and level of microplastic pollution may be more closely correlated to specific human activities in a region than to global plastic production and utilization as such.