Larval Swimming Research Articles

A behavioral and modeling study of control algorithms underlying the translational optomotor response in larval zebrafish with implications for neural circuit function.

The optomotor response (OMR) is central to the locomotory behavior in diverse animal species including insects, fish and mammals. Furthermore, the study of the OMR in larval zebrafish has become a key model system for investigating the neural basis of sensorimotor control. However, a comprehensive understanding of the underlying control algorithms is still outstanding. In fish it is often assumed that the OMR, by reducing average optic flow across the retina, serves to stabilize position with respect to the ground. Yet the degree to which this is achieved, and how it could emerge from the intermittent burst dynamics of larval zebrafish swimming, are unclear. Here, we combine detailed computational modeling with a new approach to free-swimming experiments in which we control the amount of visual feedback produced by a given motor effort by varying the height of the larva above a moving grid stimulus. We develop an account of underlying feedback control mechanisms that describes both the bout initiation process and the control of swim speed during bouts. We observe that the degree to which fish stabilize their position is only partial and height-dependent, raising questions about its function. We find the relative speed profile during bouts follows a fixed temporal pattern independent of absolute bout speed, suggesting that bout speed and bout termination are not separately controlled. We also find that the reverse optic flow, experienced when the fish is swimming faster than the stimulus, plays a minimal role in control of the OMR despite carrying most of the sensory information about self-movement. These results shed new light on the underlying dynamics of the OMR in larval zebrafish and will be crucial for future work aimed at identifying the neural basis of this behavior.

Spatial genetic differentiation correlates with species assemblage turnover across tropical reef fish lineages

AbstractAimEvaluating the similarity of diversity patterns across micro‐ to macroevolutionary scales in natural communities, such as species–genetic diversity correlations (SGDCs), may inform on processes shaping community assembly. However, whether SGDCs not only hold across communities but also across lineages has never been explored so far. Here we investigated SGDCs across co‐distributed taxa for different spatial components (α, β, γ), and formally tested the influence of dispersal traits on β‐SGDCs.LocationWestern Indian Ocean.Time period2016–2017.Major taxa studiedTropical reef fish species with contrasting dispersal traits.MethodsUsing double‐digest restriction‐site associated DNA sequencing (ddRADseq) Single Nucleotide Polymorphism data for 20 tropical reef fishes and distribution data of 2,446 species belonging to 12 families, we analysed the correlations between within‐species genetic diversity and within‐family species diversity (i.e., lineage diversity) for the three spatial components (α, β, γ‐SGDCs). We then related the strength of β‐SGDCs per species to proxies of larval dispersal abilities.ResultsWe detected positive and significant lineage‐based SGDC only for the β component, that is, the families showing the greatest level of species turnover among sites contain the species with the greatest levels of genetic differentiation. We showed that the Monsoon Drift mainly explained the β‐diversity patterns at both intraspecific and interspecific levels. Higher β‐SGDCs were found for species with short pelagic larval duration and weak larval swimming capacity.Main conclusionsOur study reveals a strong correlation between genetic and species β‐diversity, a result explained by the presence of a ‘soft’ barrier and mediated by larval dispersal processes. This suggests that vicariance and dispersal limitation are major processes shaping β‐diversity patterns from microevolutionary to macroevolutionary scales in tropical reef fishes.

Influences of brood-dependent behavioral variation on blue crab (Callinectes sapidus) larval transport in a wind-driven estuarine plume

Blue crabs (Callinectes sapidus) support valuable fisheries in the US mid-Atlantic Bight (MAB), and their unpredictable and variable recruitment suggests that a better understanding of larval development and dispersal is needed. Blue crab larval dispersal involves export to the continental shelf followed by re-entry of estuaries. Transport is facilitated by wind and buoyancy-driven surface currents, and zoeae generally maintain a near-surface distribution. Though several studies have investigated C. sapidus larval dispersal, none have evaluated the effects of behavioral variability on transport. This study simulates first-stage larvae using documented behavioral variation within an idealized wind-driven estuarine plume in order to investigate the roles of swimming behavior, stratified current systems, and turbulent mixing on transport. Model results showed that larval transport was predominately influenced by wind speed, but transport was significantly affected by behavioral characteristics. Faster swimming larvae were more able to maintain a near-surface position and had more control over their vertical position despite vertical diffusivity, and in all model scenarios, larvae travelled farther and had different vertical distributions than passive particles. Modeled net transport distance of simulated broods differed by a factor of 1.8, with a maximum 4.7 fold difference between any individual larvae, and in all scenarios behaving larvae showed further net transport and a shallower vertical distribution than passive particles. These results indicate that blue crab larval swimming ability and variability in behavioral traits may be an important factor in C. sapidus larval dispersal.

Interactive effects of Louisiana Sweet Crude (LSC) thin oil sheens and ultraviolet light on mortality and swimming behavior of the larval Eastern oyster, Crassostrea virginica

The Eastern oyster (Crassostrea virginica) is an important commercial bivalve species which also has numerous ecological roles including biogeochemical cycling, providing habitat for larval fish and crustaceans, and reducing the impacts of coastal storms. Oil may pose a threat to oyster larvae swimming in the water column, leading to potential negative effects on survival, growth, and development. Oil toxicity may be further enhanced by chemical changes in the presence of sunlight. This study determined the toxicity of thin oil sheens with and without ultraviolet (UV) light, then examined the latent effects of the short term exposure on longer term survival and swimming ability. Larval C. virginica were exposed to four different oil sheen thicknesses for 24 h with either no UV light or 2-h UV light. Following the exposure, larvae were transferred to clean seawater and no UV light for 96 h. The presence of a 2-h UV light exposure significantly increased oyster mortality, indicating photo-enhanced toxicity. The LC50 for a 24-h oil sheen exposure without UV was 7.26 µm (23 µg/L PAH50) while a 2 h-UV exposure lowered the sheen toxicity threshold to 2.67 µm (10 µg/L PAH50). A previous 24-h oil sheen exposure (≥0.5 µm) led to latent effects on larval oyster survival, regardless of previous UV exposure. Sublethal impacts to larval oyster swimming behavior were also observed from the previous oil sheen exposure combined with UV exposure. This study provides new data for the toxicity of thin oil sheens to a sensitive early life stage of estuarine bivalve.

Assessing the influence of behavioural parameterisation on the dispersal of larvae in marine systems

Predicting dispersal and quantifying ecological connectivity are increasingly referenced as fundamental to understanding how biodiversity is structured across space and time. Dispersal models can provide insight, but their predictions are influenced by our capacity to simulate the biology and physics known to influence dispersal. In a marine context, vertical swimming behaviour is considered important in influencing the spatial organisation of species across seascapes, but the mechanisms underpinning these movements remain unresolved, making it unclear how best to incorporate behaviour within models. Here, using a 3-D hydrodynamic model coupled with a Lagrangian particle tracker, we show how different modelled larval behaviours, alongside spatial and temporal hydrodynamic changes, influence larval dispersal predictions. Additionally, we compare the application of a novel approach of reverse-engineered larval swimming behaviour against two commonly modelled behaviours: passive dispersal and tidal vertical migration (TVM). We used statistical models (LME and GAM) to test the effects of change in tidal state conditions, season, and planktonic larval duration in conjunction with behavioural parameters on dispersal. For shorter PLDs (i.e., 1 day), we find that passive models match ‘behaving’ model outputs, but for longer PLDs, excluding behaviour leads to overestimates of dispersal; an effect that increases with time. Our results highlight the sensitivity of biophysical models to behavioural inputs, specifically how vertical migration behaviour can significantly reduce dispersal distance - especially for species with longer planktonic durations. This study demonstrates the disproportionate effects that even a single behaviour - vertical swimming - can have on model predictions, our understanding of ecosystem functioning, and ultimately, the ecological coherence of marine systems.

Turbulence affects larval vertical swimming in the cold-water coral Lophelia pertusa

Vertical migration of marine larvae may drastically affect their dispersal, especially if they are spawned in the deep sea. Previous studies have shown that the planktonic larvae of the cold-water coral Lophelia pertusa in still water swim upwards at a speed of ca. 0.5 mm s-1 during a pre-competency period of 3–5 weeks. This behavioral trait is thought to benefit dispersion of larvae as it promotes near surface drift in relatively strong currents. In the ocean however, larvae regularly encounter turbulent water movements potentially impeding their swimming ability. With no apparent stabilizing mechanism, it may be expected that the body orientation of these larvae, and consequently their directed swimming, is sensitive to perturbation by external forces. We investigated the effects of turbulence on vertical swimming of pre-competent L. pertusa larvae by exposing them to relevant turbulence intensities within a grid-stirred tank. Larval movement and water flow were simultaneously recorded, allowing for analysis of individual larval swimming velocities. We showed that the upwards directed swimming speed generally decreased with increasing turbulence, dropping to non-significant in turbulence levels occurring near ocean boundaries. Our results do however suggest that L. pertusa larvae maintain their upwards directed swimming, albeit at reduced speed, in a major part of the water column, thus allowing them to spend part of their planktonic phase in the uppermost ocean layer. This new insight into the behavior of L. pertusa larvae in their natural environment strengthens the notion of the species as one with strong potential for long-distance dispersal. Such information is important for the understanding of L. pertusa population connectivity, and vital when developing tools for modelling of larval transport.

Applying behavioral studies to the ecotoxicology of corals: A case study on Acropora millepora

Behavioral responses are considered sensitive and effective indicators of organism stress. As the demand for standardized coral toxicity tests grows, innovative tools that allow for automatic and quantitative measurements of these behaviors may complement ecotoxicological studies. The rapid growth of competitive marine algae in aquaculture systems is a major issue for generating coral spat for reef restoration, and the application of non-toxic antifouling (AF) coatings might effectively mitigate this issue. While these coatings do not appear to be toxic to sensitive coral larvae, their potential to affect larval mobility has not been tested. In this context, we tested the effect of three recently-developed and potentially non-toxic AF coatings: (i) antiadhesive, (ii) cerium dioxide (CeO2-x) nanoparticle, and (iii) encapsulated biocide dichlorooctylisothiazolinone (DCOIT) on the swimming velocity and activity of Acropora millepora coral larvae for potential use in reef-restoration activities. The behavior of 32 coral larvae per AF treatment were recorded, each for 25 min, in a self-constructed dark box with two camera recording sets in parallel. The tracking analysis was performed with the software Noldus EthoVision XT. The mean larval swimming velocity on control tiles of 93.1 ± 5.6 mm min-1 (and activity of 62.8 ± 5.2%) was nearly 2-fold faster (higher) than on the antiadhesive, (CeO2-x) nanoparticle and DCOIT coatings, respectively. Larvae exposed to the DCOIT-coated tiles remained almost stationary. Although the underlying cause and consequence of these results require further investigation, tracking of coral larval swimming behavior was identified as a reliable and feasible method for assessing potential non-lethal responses to AF coatings. As changes in behavior could have significant consequences for larval survival and settlement, they are important endpoints to consider, and the quantification of behavioral responses may be a meaningful and sensitive tool. Therefore, we recommend the use of behavioral studies for coral larval assessments in ecotoxicology as a valuable endpoint. For methodological standardization and implementation, our study also features a detailed guide for video-processing and track analysis of A. millepora coral larvae in EthoVision.

Light spectrum impacts on development respiratory metabolism and antioxidant capacity of larval swimming crab Portunus trituberculatus

The effects of the spectrum on the development, respiratory metabolism, and antioxidant capacity of the larval swimming crab Portunus trituberculatus were studied. Seven light spectra, i.e., purple (400 nm), blue (425 nm), cyan (510 nm), green (525 nm), yellow (598 nm), red (638 nm), and white (full spectrum), were estimated. The larvae had the optimum survival rate and development under cyan light. On the contrary, larvae in red and yellow lights had poor growth performance. The oxygen consumption rate (OCR) dropped while the ammonia excretion rate (AER) rose as the larvae developed. Early larvae’s oxygen-nitrogen ratio (O: N) fell when exposed to red light, suggesting more protein was utilized in the respiratory process. Regarding the antioxidant system, crab had the lowest malondialdehyde (MDA) under green, cyan and yellow light, and the highest total antioxidant capacity (T-AOC) in cyan light. Taken together, the current results suggest that cyan was the optimum spectrum for the development of P. trituberculatus larvae.

Heritability of dispersal-related larval traits in the clown anemonefish Amphiprion percula.

A major goal of marine ecology is to identify the drivers of variation in larval dispersal. Larval traits are emerging as an important potential source of variation in dispersal outcomes, but little is known about how the evolution of these traits might shape dispersal patterns. Here, we consider the potential for adaptive evolution in two possibly dispersal-related traits by quantifying the heritability of larval size and swimming speed in the clown anemonefish (Amphiprion percula). Using a laboratory population of wild-caught A. percula, we measured the size and swimming speed of larvae from 24 half-sibling families. Phenotypic variance was partitioned into genetic and environmental components using a linear mixed-effects model. Importantly, by including half-siblings in the breeding design, we ensured that our estimates of genetic variance do not include nonheritable effects shared by clutches of full-siblings, which could lead to significant overestimates of heritability. We find unequivocal evidence for the heritability of larval body size (estimated between 0.21 and 0.34) and equivocal evidence for the heritability of swimming speed (between 0.05 and 0.19 depending on the choice of prior). From a methodological perspective, this work demonstrates the importance of evaluating sensitivity to prior distribution in Bayesian analysis. From a biological perspective, it advances our understanding of potential dispersal-related larval traits by quantifying the extent to which they can be inherited and thus have the potential for adaptive evolution.

Behavioral and transcriptomic changes in butenolide treated larvae of the cosmopolitan fouling bryozoan Bugulina (Bugula) neritina

The arborescent bryozoanBugulina neritinais a cosmopolitan fouling species in sub-tropical to sub-temperate waters. The butenolide compound 5-octylfuran-2(5H)-one (hereafter named butenolide) reportedly inhibits larval settlement ofB. neritina, but its effect on the larval behavioral and transcriptomic responses remained unclear. We report that 10 μg mL-1or higher concentration of butenolide and/or prolonged treatment (10 h or longer) resulted in significant increase in larval mortality and prominent spiral larval swimming behavior. The transcriptomic analyses not only revealed up-regulation of typical stress-related protein genes in response to 10 mg mL-1butenolide treatment, but also indicated up-regulation ofmucin, synaptic genes and nitric oxide signaling genes. In addition, the expression of developmental genes and lipid biosynthesis genes were also affected. Overall, our larval behavior and transcriptome analyses reflected the impact of butenolide on the metabolism, neuronal and molecular signaling inB. neritinalarvae.

Exposure of gametes to aged nano-sized plastic particles during fertilization can influence early larval development in the European whitefish (Coregonus lavaretus)

Plastic pollution has been a growing environmental concern for decades, increasingly affecting both marine and freshwater ecosystems worldwide. Nano-sized plastic particles (NPs) potentially have various toxicological impacts on aquatic organisms and the ecosystem; however, less is known about their possible adverse effects on the reproductive biology and offspring traits of fishes. The present study investigated whether an acute exposure of gametes to aged NPs during fertilization affects offspring early mortality, hatching time, body size at hatching or swimming performance of larvae in a common freshwater fish, the European whitefish (Coregonus lavaretus). Using a replicated full-factorial breeding design, we fertilized the eggs of seven females with the milt of seven males both under exposure medium containing aged 270 nm polystyrene NPs and under control medium. In comparison with the control group, exposure of gametes to NPs increased larval body length slightly but significantly, whereas the embryo mortality, hatching time, and larval swimming performance were not affected. Maternal identity affected significantly all the studied offspring traits while paternal identity only affected the offspring length. Our results suggest that the studied acute exposure of gametes to aged NPs might have interfered normal embryonic development by affecting larval size, but this did not seemingly compromise offspring performance.

Photoperiod affects the survival rate but not the development of larval swimming crab Portunus trituberculatus

Photoperiod affects the survival rate but not the development of larval swimming crab Portunus trituberculatus

Environmental microplastics disrupt swimming activity in acute exposure in Danio rerio larvae and reduce growth and reproduction success in chronic exposure in D. rerio and Oryzias melastigma

Microplastics (MPs), widely present in aquatic ecosystems, can be ingested by numerous organisms, but their toxicity remains poorly understood. Toxicity of environmental MPs from 2 beaches located on the Guadeloupe archipelago, Marie Galante (MG) and Petit-Bourg (PB) located near the North Atlantic gyre, was evaluated. A first experiment consisted in exposing early life stages of zebrafish (Danio rerio) to MPs at 1 or 10 mg/L. The exposure of early life stages to particles in water induced no toxic effects except a decrease in larval swimming activity for both MPs exposures (MG or PB). Then, a second experiment was performed as a chronic feeding exposure over 4 months, using a freshwater fish species, zebrafish, and a marine fish species, marine medaka (Oryzias melastigma). Fish were fed with food supplemented with environmentally relevant concentrations (1% wet weight of MPs in food) of environmental MPs from both sites. Chronic feeding exposure led to growth alterations in both species exposed to either MG or PB MPs but were more pronounced in marine medaka. Ethoxyresorufin-O-deethylase (EROD) and acetylcholinesterase (AChE) activities were only altered for marine medaka. Reproductive outputs were modified following PB exposure with a 70 and 42% decrease for zebrafish and marine medaka, respectively. Offspring of both species (F1 generation) were reared to evaluate toxicity following parental exposure on unexposed larvae. For zebrafish offspring, it revealed premature mortality after parental MG exposure and parental PB exposure produced behavioural disruptions with hyperactivity of F1 unexposed larvae. This was not observed in marine medaka offspring. This study highlights the ecotoxicological consequences of short and long-term exposures to environmental microplastics relevant to coastal marine areas, which represent essential habitats for a wide range of aquatic organisms.

Electroporation-Based CRISPR/Cas9 Mosaic Mutagenesis of β-Tubulin in the Cultured Oyster

Genome editing using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 is enabling genetics improvement of productive traits in aquaculture. Previous studies have proven CRISPR/Cas9 to be feasible in oyster, one of the most cultured shellfish species. Here, we applied electroporation-based CRISPR/Cas9 knockout of β-tubulin and built a highly efficient genome editing system in Crassostrea gigas angulate. We identified the β-tubulin gene in the oyster genome and showed its spatiotemporal expression patterns by analyzing RNA-seq data and larval in situ hybridization. We further designed multiple highly specific guide RNAs (sgRNAs) for its coding sequences. Long fragment deletions were detected in the mutants by agarose gel electrophoresis screening and further verified by Sanger sequencing. In addition, the expression patterns of Cgβ-tubulin in the trochophore peritroch and intestinal cilia cells were altered in the mutants. Scanning electron microscopy represented shortened and almost complete depleted cilia at the positions of peritroch and the posterior cilium ring in Cgβ-tubulin mosaic knockout trochophores. Moreover, the larval swimming behavior in the mutants was detected to be significantly decreased by motility assay. These results demonstrate that β-tubulin is sufficient to mediate cilia development and swimming behavior in oyster larvae. By applying Cgβ-tubulin as a marker gene, our study established CRISPR/Cas9-mediated mosaic mutagenesis technology based on electroporation, providing an efficient tool for gene function validation in the oyster. Moreover, our research also set up an example that can be used in genetic engineering breeding and productive traits improvement in oysters and other aquaculture species.

The Effects of Anthropogenic Electromagnetic Fields (EMF) on the Early Development of Two Commercially Important Crustaceans, European Lobster, Homarus gammarus (L.) and Edible Crab, Cancer pagurus (L.)

Proposed offshore windfarm sites could overlap with the brooding and spawning habitats of commercially important crustacea, including European lobster, Homarus gammarus and Edible crab, Cancer pagurus. Concerns have been raised on the biological effects of Electromagnetic Fields (EMFs) emitted from subsea power cables on the early life history of these species. In this study, ovigerous female H. gammarus and C. pagurus were exposed to static (Direct Current, DC) EMFs (2.8 mT) throughout embryonic development. Embryonic and larval parameters, deformities, and vertical swimming speed of freshly hatched stage I lobster and zoea I crab larvae were assessed. EMF did not alter embryonic development time, larval release time, or vertical swimming speed for either species. Chronic exposure to 2.8 mT EMF throughout embryonic development resulted in significant differences in stage-specific egg volume and resulted in stage I lobster and zoea I crab larvae exhibiting decreased carapace height, total length, and maximum eye diameter. An increased occurrence of larval deformities was observed in addition to reduced swimming test success rate amongst lobster larvae. These traits may ultimately affect larval mortality, recruitment and dispersal. This study increases our understanding on the effects of anthropogenic, static EMFs on crustacean developmental biology and suggests that EMF emissions from subsea power cables could have a measurable impact on the early life history and consequently the population dynamics of H. gammarus and C. pagurus.

Multimodal Characterization of Seizures in Zebrafish Larvae.

Epilepsy accounts for a significant proportion of the world’s disease burden. Indeed, many research efforts are produced both to investigate the basic mechanism ruling its genesis and to find more effective therapies. In this framework, the use of zebrafish larvae, owing to their peculiar features, offers a great opportunity. Here, we employ transgenic zebrafish larvae expressing GCaMP6s in all neurons to characterize functional alterations occurring during seizures induced by pentylenetetrazole. Using a custom two-photon light-sheet microscope, we perform fast volumetric functional imaging of the entire larval brain, investigating how different brain regions contribute to seizure onset and propagation. Moreover, employing a custom behavioral tracking system, we outline the progressive alteration of larval swim kinematics, resulting from different grades of seizures. Collectively, our results show that the epileptic larval brain undergoes transitions between diverse neuronal activity regimes. Moreover, we observe that different brain regions are progressively recruited into the generation of seizures of diverse severity. We demonstrate that midbrain regions exhibit highest susceptibility to the convulsant effects and that, during periods preceding abrupt hypersynchronous paroxysmal activity, they show a consistent increase in functional connectivity. These aspects, coupled with the hub-like role that these regions exert, represent important cues in their identification as epileptogenic hubs.

Altered Larval Yellow Perch Swimming Behavior Due to Methylmercury and PCB126 Detected Using Hidden Markov Chain Models.

Fish swimming behavior is a commonly measured response in aquatic ecotoxicology because behavior is considered a whole organism-level effect that integrates many sensory systems. Recent advancements in animal behavior models, such as hidden Markov chain models (HMM), suggest an improved analytical approach for toxicology. Using both new and traditional approaches, we examined the sublethal effects of PCB126 and methylmercury on yellow perch (YP) larvae (Perca flavescens) using three doses. Both approaches indicate larvae increase activity after exposure to either chemical. The middle methylmercury-dosed larvae showed multiple altered behavior patterns. First, larvae had a general increase in activity, typically performing more behavior states, more time swimming, and more swimming bouts per second. Second, when larvae were in a slow or medium swimming state, these larvae tended to switch between these states more often. Third, larvae swam slower during the swimming bouts. The upper PCB126-dosed larvae exhibited a higher proportion and a fast swimming state, but the total time spent swimming fast decreased. The middle PCB126-dosed larvae transitioned from fast to slow swimming states less often than the control larvae. These results indicate that developmental exposure to very low doses of these neurotoxicants alters YP larvae overall swimming behaviors, suggesting neurodevelopment alteration.

Tropical larval and juvenile fish critical swimming speed (U-crit) and morphology data

Fish swimming capacity is a key life history trait critical to many aspects of their ecology. U-crit (critical) swimming speeds provide a robust, repeatable relative measure of swimming speed that can serve as a useful surrogate for other measures of swimming performance. Here we collate and make available one the most comprehensive datasets on U-crit swimming abilities of tropical marine fish larvae and pelagic juveniles, most of which are reef associated as adults. The dataset includes U-crit speed measurements for settlement stage fishes across a large range of species and families obtained mostly from field specimens collected in light traps and crest nets; and the development of swimming abilities throughout ontogeny for a range of species using reared larvae. In nearly all instances, the size of the individual was available, and in many cases, data include other morphological measurements (e.g. “propulsive area”) useful for predicting swimming capacity. We hope these data prove useful for further studies of larval swimming performance and other broader syntheses.

Planktonic sea urchin larvae change their swimming direction in response to strong photoirradiation.

To survive, organisms need to precisely respond to various environmental factors, such as light and gravity. Among these, light is so important for most life on Earth that light-response systems have become extraordinarily developed during evolution, especially in multicellular animals. A combination of photoreceptors, nervous system components, and effectors allows these animals to respond to light stimuli. In most macroscopic animals, muscles function as effectors responding to light, and in some microscopic aquatic animals, cilia play a role. It is likely that the cilia-based response was the first to develop and that it has been substituted by the muscle-based response along with increases in body size. However, although the function of muscle appears prominent, it is poorly understood whether ciliary responses to light are present and/or functional, especially in deuterostomes, because it is possible that these responses are too subtle to be observed, unlike muscle responses. Here, we show that planktonic sea urchin larvae reverse their swimming direction due to the inhibitory effect of light on the cholinergic neuron signaling>forward swimming pathway. We found that strong photoirradiation of larvae that stay on the surface of seawater immediately drives the larvae away from the surface due to backward swimming. When Opsin2, which is expressed in mesenchymal cells in larval arms, is knocked down, the larvae do not show backward swimming under photoirradiation. Although Opsin2-expressing cells are not neuronal cells, immunohistochemical analysis revealed that they directly attach to cholinergic neurons, which are thought to regulate forward swimming. These data indicate that light, through Opsin2, inhibits the activity of cholinergic signaling, which normally promotes larval forward swimming, and that the light-dependent ciliary response is present in deuterostomes. These findings shed light on how light-responsive tissues/organelles have been conserved and diversified during evolution.

High CO2 inhibits substratum exploration and settlement of coral larvae

Biological and physical factors affecting coral recruitment are critical in influencing the recovery of coral communities after disturbance. While ocean acidification (OA) can reduce coral settlement and the early growth of coral recruits, the impact of OA on coral larval swimming behavior is unknown. Here, we investigated the effects of elevated CO2 on the swimming behavior and settlement of coral larvae of 2 common Acropora species. Larvae were exposed to 4 CO2 partial pressure (pCO2) conditions consistent with the current Intergovernmental Panel for Climate Change predictions for the next few centuries (pCO2: 393, 853, 1485 and 3022 µatm; pH: 8.1, 7.8, 7.6 and 7.3) in 2 laboratory experiments. We found that bottom exploration, expressed as the proportion of A. cytherea and A. pulchra larvae present in the bottom part of experimental cylinders, decreased by 92 and 98%, respectively, from the ambient to highest CO2 treatment. When offered the choice to settle on the crustose coralline algae Titanoderma prototypum, a well-known positive settlement cue, the percentage of larvae that settled on the fragments declined rapidly as pCO2 increased, with no larvae settling in the highest CO2 treatment. These results suggest that OA may negatively affect coral recruitment via direct effects on larval swimming behavior, with larvae avoiding benthic probing in response to high CO2.