Neuropeptide Modulation Enables Biphasic Internetwork Coordination via a Dual-Network Neuron.

Linked rhythmic behaviors, such as respiration/locomotion or swallowing/chewing, often require coordination for proper function. Despite its prevalence, the cellular mechanisms controlling coordination of the underlying neural networks remain undetermined in most systems. We use the stomatogastric nervous system of the crab Cancer borealis to investigate mechanisms of internetwork coordination, due to its small, well-characterized feeding-related networks (gastric mill [chewing, ∼0.1 Hz]; pyloric [filtering food, ∼1 Hz]). Here, we investigate coordination between these networks during the Gly1-SIFamide neuropeptide modulatory state. Gly1-SIFamide activates a unique triphasic gastric mill rhythm in which the typically pyloric-only LPG neuron generates dual pyloric-plus gastric mill-timed oscillations. Additionally, the pyloric rhythm exhibits shorter cycles during gastric mill rhythm-timed LPG bursts, and longer cycles during IC, or IC plus LG gastric mill neuron bursts. Photoinactivation revealed that LPG is necessary to shorten pyloric cycle period, likely through its rectified electrical coupling to pyloric pacemaker neurons. Hyperpolarizing current injections demonstrated that although LG bursting enables IC bursts, only gastric mill rhythm bursts in IC are necessary to prolong the pyloric cycle period. Surprisingly, LPG photoinactivation also eliminated prolonged pyloric cycles, without changing IC firing frequency or gastric mill burst duration, suggesting that pyloric cycles are prolonged via IC synaptic inhibition of LPG, which indirectly slows the pyloric pacemakers via electrical coupling. Thus, the same dual-network neuron directly conveys excitation from its endogenous bursting and indirectly funnels synaptic inhibition to enable one network to alternately decrease and increase the cycle period of a related network.

Future redistribution of fishery resources suggests biological and economic trade-offs according to the severity of the emission scenario.

Climate change is anticipated to have long-term and pervasive effects on marine ecosystems, with cascading consequences to many ocean-reliant sectors. For the marine fisheries sector, these impacts can be further influenced by future socio-economic and political factors. This raises the need for robust projections to capture the range of potential biological and economic risks and opportunities posed by climate change to marine fisheries. Here, we project future changes in the abundance of eight commercially important fish and crab species in the eastern Bering Sea and Chukchi Sea under different CMIP6 Shared Socioeconomic Pathways (SSPs) leading to contrasting future (2021-2100) scenarios of warming, sea ice concentration, and net primary production. Our results revealed contrasting patterns of abundance and distribution changes across species, time periods and climate scenarios, highlighting potential winners and losers under future climate change. In particular, the least changes in future species abundance and distribution were observed under SSP126. However, under the extreme scenario (SSP585), projected Pacific cod and snow crab abundances increased and decreased, respectively, with concurrent zonal and meridional future shifts in their centers of gravity. Importantly, projected changes in species abundance suggest that fishing at the same distance from the current major port in the Bering Sea (i.e., Dutch Harbor) could yield declining catches for highly valuable fisheries (e.g., Pacific cod and snow crab) under SSP585. This is driven by strong decreases in future catches of highly valuable species despite minimal declines in maximum catch potential, which are dominated by less valuable taxa. Hence, our findings show that projected changes in abundance and shifting distributions could have important biological and economic impacts on the productivity of commercial and subsistence fisheries in the eastern Bering and Chukchi seas, with potential implications for the effective management of transboundary resources.

Metabolic scaling of an invasive mussel depends on temperature and chemical cues from an invasive predator.

Metabolism drives various biological processes, potentially influencing the ecological success and evolutionary fitness of species. Understanding diverse metabolic rates is fundamental in biology. Mechanisms underlying adaptation to factors like temperature and predation pressure remain unclear. Our study explored the role of temperature and predation pressure in shaping the metabolic scaling of an invasive mussel species (Brachidontes pharaonis). Specifically, we performed laboratory-based experiments to assess the effects of phenotypic plasticity on the metabolic scaling by exposing the mussels to water conditions with and without predator cues from another invasive species (the blue crab, Callinectes sapidus) across various temperature regimes. We found that temperature effects on metabolic scaling of the invasive mussels are mediated by the presence of chemical cues of an invasive predator, the blue crab. Investigating temperature-predator interactions underscores the importance of studying the ecological effects of global warming. Our research advances our understanding of how environmental factors jointly impact physiological processes.

Fiddler crabs from highly disturbed beaches are more sensitive to human presence.

The presence of humans frequently modifies the behavior of animals, particularly their foraging patterns, compromising energetic demands. The fiddler crab Leptuca leptodactyla inhabits mangroves with high degrees of anthropogenic influence. Thus, we tested if populations living in highly anthropized mangroves respond differently from those living in more protected areas. We predict that individuals from touristy areas will be more tolerant to humans and will resume their activities sooner after disturbance. To do so, we conducted an experiment that consisted in the approach of an observer to the burrows, recording the response of individuals to the stimuli. The experiment took place in July 2022, in Ubatuba, São Paulo, Brazil. We analysed the duration and latency of various behaviors of a total of 80 adult males from two populations (high and low anthropogenic influence). Contrary to our predictions, individuals from the anthropized population were less tolerant, spending more time inside their burrows and taking longer to resume their activities. Therefore, fiddler crabs were not habituated to human presence. These results help us understand the learning process in invertebrates and their ability to select stimuli, contributing to understanding the impacts of human-wildlife interactions.

The tricarboxylic acid cycle is inhibited under acute stress from carbonate alkalinity in the gills of Eriocheir sinensis.

Owing to population growth and environmental pollution, freshwater aquaculture has been rapidly shrinking in recent years. Aquaculture in saline-alkaline waters is a crucial strategy to meet the increasing demand for aquatic products. The Chinese mitten crab is an important economic food in China, but the molecular mechanism by which it tolerates carbonate alkalinity (CA) in water remains unclear. Here, we found that enzyme activities of the tricarboxylic acid (TCA) cycle in the gills, such as citrate synthase, isocitrate dehydrogenase, α-ketoglutarate dehydrogenase, and malate dehydrogenase, were markedly reduced under CA stress induced by 40 mM NaHCO3. Secondly, the TCA cycle in the gills is inhibited under acute CA stress, according to proteomic and metabolomic analyses. The expressions of six enzymes, namely aconitate hydratase, isocitrate dehydrogenase, 2-oxoglutarate dehydrogenase, dihydrolipoyl dehydrogenase, succinate-CoA ligase, and malate dehydrogenase, were downregulated, resulting in the accumulation of phosphoenolpyruvic acid, citric acid, cis-aconitate, and α-ketoglutaric acid. Finally, we testified that if the TCA cycle is disturbed by malonate, the survival rate increases in CA water. To our knowledge, this is the first study to show that the TCA cycle in the gills is inhibited under CA stress. Overall, the results provide new insights into the molecular mechanism of tolerance to saline-alkaline water in crabs, which helped us expand the area for freshwater aquaculture and comprehensively understand the physiological characteristics of crab migration.

Climatic resilience: Marine heatwaves do not influence the variations of green crab (Carcinus maenas) megalopae supply patterns to a Western Iberian estuary.

Extreme climatic events like marine heatwaves (MHWs) are becoming more frequent, intense, and longer lasting all around the world. The consequences of these anomalously warm periods are devastating for marine ecosystems. Still, little is known about these extreme events off the western Iberia coast. Here we analyzed MHW events occurring from 1982 to 2020 on the Aveiro coast, western Iberia coast of Portugal. A total of 79 events were detected for the region, with an average duration of 15.8 days, and a mean intensity of 1.9 °C ± 0.4 °C above the 90th percentile of sea surface temperatures (SST) for the region. The maximum intensity of the events has increased by 0.5 °C over the last decade. The relation between SST, and therefore, MHW events, the North Atlantic Oscillation index (NAO), and the regional Iberian Upwelling Index (UI) was identified. The intense upwelling of the region seems to mitigate the duration of warming conditions, resulting in shorter MHW events. Furthermore, the impacts of SST and MHW events on the supply patterns of Carcinus maenas megalopae were examined, utilizing daily data from 2002, 2006-2009, 2012, and 2013, collected at the entrance of Ria de Aveiro. Cross-correlations were employed to assess the effect of SST on megalopae supply, while ordinary least square cumulative sums were used to identify variations over time. The influence of SST on supply was noticed with a 5-to-11-day lag, but this relation changed over the years. Contrary to our hypothesis, we found no evidence supporting a diminishment in megalopae supply due to MHW events. These elusive findings, coupled with the apparent lack of influence of these extreme events, highlight the relatively weak intensity and brief duration of the MHW events in the region, coupled with the high thermal tolerance of these species.

Mitogenome selection shaped the terrestrial adaptation of Grapsidae (Decapoda: Brachyura).

The colonization of aquatic to terrestrial habitats by brachyuran crabs requires genetic innovations as well as morphological adaptations to adapt to terrestrial environments. The genetic basis of such adaptive evolution, however, is largely unknown. This study focuses on terrestrialization in Geograpsus (Grapsidae) the only highly terrestrial genus in this family, which represents a notable example of terrestrial adaptive radiation. Here, we sequenced the mitogenomes of two Geograpsus species and used the mitogenomes of 215 representative crabs to construct phylogenetic and time frameworks that we used to infer terrestrial origins and evolution. Using mitochondrial genomic data, we demonstrated that marine crab ancestors began to settle on land during the early Eocene. Ocean acidification, the Paleocene-Eocene Thermal Maximum (PETM), and mangrove expansion at that time may have driven the diversification and ecological expansion of these terrestrial crabs. Evolutionary analyses reveal strong positive selection signals on monophyletic lineages of Grapsidae, especially the terrestrial species of Geograpsus. Positively selected sites in functionally important regions of ND5 and ND4 may imply enhanced energy metabolism in Grapsidae compared to other crabs, and may have played an important role in their terrestrial adaptation. Overall, our work provides valuable resources and opportunities to reveal the adaptation of crabs to complex terrestrial environments.

Mesocosm study of PAC-modified clay effects on Karenia brevis cells and toxins, chemical dynamics, and benthic invertebrate physiology.

Modified clay compounds are used globally as a method of controlling harmful algal blooms, and their use is currently under consideration to control Karenia brevis blooms in Florida, USA. In 1400 L mesocosm tanks, chemical dynamics and lethal and sublethal impacts of MC II, a polyaluminum chloride (PAC)-modified kaolinite clay, were evaluated over 72 h on a benthic community representative of Sarasota Bay, which included blue crab (Callinectes sapidus), sea urchin (Lytechinus variegatus), and hard clam (Mercenaria campechiensis). In this experiment, MC II was dosed at 0.2 g L-1 to treat bloom-level densities of K. brevis at 1 × 106 cells L-1. Cell removal in MC II-treated tanks was 57% after 8 h and 95% after 48 h. In the water column, brevetoxin analogs BTx-1 and BTx-2 were found to be significantly higher in untreated tanks at 24 and 48 h, while in MC II-treated tanks, BTx-3 was found to be higher at 48 h and BTx-B5 was found to be higher at 24 and 48 h. In MC II floc, we found no significant differences in BTx-1 or BTx-2 between treatments for any time point, while BTx-3 was found to be significantly higher in the MC II-treated tanks at 48 and 72 h, and BTx-B5 was higher in MC II-treated tanks at 24 and 72 h. Among various chemical dynamics observed, it was notable that dissolved phosphorus was consistently significantly lower in MC II tanks after 2 h, and that turbidity in MC II tanks returned to control levels 48 h after treatment. Dissolved inorganic carbon and total seawater alkalinity were significantly reduced in MC II tanks, and partial pressure of CO2 (pCO2) was significantly higher in the MC II-only treatment after 2 h. In MC II floc, particulate phosphorus was found to be significantly higher in MC II tanks after 24 h. In animals, lethal and sublethal responses to MC II-treated K. brevis did not differ from untreated K. brevis for either of our three species at any time point, suggesting MC II treatment at this dosage has negligible impacts to these species within 72 h of exposure. These results appear promising in terms of the environmental safety of MC II as a potential bloom control option, and we recommend scaling up MC II experiments to field trials in order to gain deeper understanding of MC II performance and dynamics in natural waters.

Invasive European green crab (Carcinus maenas) predation in a Washington State estuary revealed with DNA metabarcoding.

Predation by invasive species can threaten local ecosystems and economies. The European green crab (Carcinus maenas), one of the most widespread marine invasive species, is an effective predator associated with clam and crab population declines outside of its native range. In the U.S. Pacific Northwest, green crab has recently increased in abundance and expanded its distribution, generating concern for estuarine ecosystems and associated aquaculture production. However, regionally-specific information on the trophic impacts of invasive green crab is very limited. We compared the stomach contents of green crabs collected on clam aquaculture beds versus intertidal sloughs in Willapa Bay, Washington, to provide the first in-depth description of European green crab diet at a particularly crucial time for regional management. We first identified putative prey items using DNA metabarcoding of stomach content samples. We compared diet composition across sites using prey presence/absence and an index of species-specific relative abundance. For eight prey species, we also calibrated metabarcoding data to quantitatively compare DNA abundance between prey taxa, and to describe an 'average' green crab diet at an intertidal slough versus a clam aquaculture bed. From the stomach contents of 61 green crabs, we identified 54 unique taxa belonging to nine phyla. The stomach contents of crabs collected from clam aquaculture beds were significantly different from the stomach contents of crabs collected at intertidal sloughs. Across all sites, arthropods were the most frequently detected prey, with the native hairy shore crab (Hemigrapsus oregonensis) the single most common prey item. Of the eight species calibrated with a quantitative model, two ecologically-important native species-the sand shrimp (Crangon franciscorum) and the Pacific staghorn sculpin (Leptocottus armatus)-had the highest average DNA abundance when detected in a stomach content sample. In addition to providing timely information on green crab diet, our research demonstrates the novel application of a recently developed model for more quantitative DNA metabarcoding. This represents another step in the ongoing evolution of DNA-based diet analysis towards producing the quantitative data necessary for modeling invasive species impacts.

No man's land support the endemic Red Sea ghost crab (Ocypode saratan) in the Gulf of Eilat.

Tourism pressure on the Red Sea ecosystem have posed significant threats to numerous endemic species, including the Ghost Crab Ocypode saratan, which is exclusively found along a small stretch of beach in the Eilat/Aqaba Red Sea Gulf. Due to the limited understanding of their ecology, we investigated how tourism impacts the behavior of this species. Employing a natural setup, we compared burrow dimensions, pyramid structures, and density across three distinct beach sections subjected to varying levels of human interference. Access to a secluded beach, referred to as "No Man's Land," provided a crucial control for our study. This facilitated a comparative analysis of ghost crab activity among beaches experiencing differing levels of human disturbances: (1) a tourist beach characterized by continual high disturbance, (2) a naval beach subject to moderate and sporadic disturbances, and (3) the isolated "no man's land" beach devoid of human presence. Our observations revealed notable differences in ghost crab density among the three beaches. Furthermore, we observed that on the secluded beach, larger individuals tended to establish burrows farther from the waterline and construct taller sand pyramids. Given the significance of sexual selection processes, their conservation becomes imperative for the survival and potential expansion of the ghost crab population across the Gulf of Eilat/Aqaba. We propose a straight-forward and cost-effective strategy: the designation of short, secluded beach enclaves along this gulf. We believe that this approach will mitigate adverse impacts of tourisms while simultaneously benefiting various sandy beach species.

Feces and molting as microplastic sinks in a mangrove crab.

We exposed adult individuals of the sentinel mangrove crab Minuca rapax to waterborne microplastics (MP; 53-63 µm polyethylene spheres) in a long-term experiment (56 days). Weassessed 1) MP effects on growth, survival, and food intake. and 2) the MP tissue acumulation and its reduction of body burden through feces and molting. MP exposure did not affect growth and survival. The hepatopancreas accumulated more MP than the gills and muscle. Most of the ingested MP particles were released in the feces and molts, indicating a rapid passage through the digestive tract. MP impaired food intake of M. rapax, with unknown consequences to the local populations. These results provide insights on MP translocation mechanisms, its elimination and toxicity associated with MP.

Differential effects of oxytetracycline on detoxification and antioxidant defense in the hepatopancreas and intestine of Chinese mitten crab under cadmium stress.

This study aims to evaluate the effects of oxytetracycline (OTC) on detoxification and oxidative defense in the hepatopancreas and intestine of Chinese mitten crab (Eriocheir sinensis) under cadmium (Cd) stress. The crab was exposed to 0.6 µM Cd, 0.6 µM OTC, and 0.6 µM Cd plus 0.6 µM OTC for 42 days. Our results showed that in the intestine, OTC alone enhanced protein carboxylation (PC) and malondialdehyde (MDA) contents, which was associated with the increased OTC accumulation. Compared to Cd alone, Cd plus OTC increased Cd and OTC contents, and reduced detoxification (i.e., glutathione (GSH) content, gene expressions of cytochrome P450 (CYP) isoforms, 7-ethoxyresorufin O-deethylase (EROD) activity, mRNA levels and activities of glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione-S-transferase (GST)), and antioxidant defense (i.e., gene expressions and activities of catalase (CAT) and superoxide dismutase (SOD)) in the intestine, leading to the increased in PC and MDA contents, suggesting that OTC had a synergistic effect on Cd-induced oxidative damage. In the hepatopancreas, although OTC alone increased OTC accumulation, it did not affect PC and MDA contents. Compared to Cd alone, Cd plus OTC reduced MDA content, which was closely related to the improvement of detoxification (i.e., GSH content, mRNA levels of CYP isoforms, EROD activity, gene expressions and activities of GPx, GR and GST), and antioxidant defense (gene expressions and activities of CAT and SOD, metallothionein content). Aryl hydrocarbon receptor (AhR) and nuclear factor E2-related factor 2 (Nrf2) transcriptional expressions were positively correlated with most detoxification- and antioxidant-related gene expressions, respectively, indicating that AhR and Nrf2 were involved in the regulation of these gene expressions. Our results unambiguously demonstrated that OTC had tissue-specific effects on Cd-induced toxicological effect in E. sinensis, which contributed to accurately evaluating Cd toxicity modulated by TCs in crab.

Landmark use by ghost crab (Ocypode quadrata) during wayfinding in a complex maze.

Species of crab have been shown to spatially track and navigate to consequential locations through different processes, such as path integration and landmark orienting. Few investigations examine their ability to wayfind in complex environments, like mazes, with multiple intersections and how they may utilize specific features to benefit this process. Spatial learning potentially would lend a fitness advantage to animals living in complicated habitats, and ghost crab (Ocypode quadrata) is a semiterrestrial species that typically occupies extensive beach environments, which present many navigational challenges. Despite their potential, there are currently no studies that investigate forms of spatial cognition in these animals. To better diversify our knowledge of this trait, the current research exposed ghost crab to a maze with seven intersections. Animals were given multiple trials to learn the location of a reward destination to a specific criterion proficiency. In one condition several landmarks were distributed throughout the maze, and in another the environment was completely empty. Results showed that ghost crab in the landmark present group were able to learn the maze faster, they required significantly fewer trials to reach the learning criterion than those in the landmark absent group. However, only approximately half of the total sample met the learning criterion, indicating the maze was rather difficult. These findings are interpreted through theories of route learning that suggest animals may navigate by establishing landmark-turn associations. Such processes have implications for the cognitive ability of ghost crab, and spatial learning in this species may support the notion of convergent evolution for this trait.

Comparative Proteomics Elucidates the Potential Mechanism of Sperm Capacitation of Chinese Mitten Crabs (Eriocheir sinensis).

Sperm capacitation is broadly defined as a suite of biochemical and biophysical changes resulting from the acquisition of fertilization ability. To gain insights into the regulation mechanism of crustacean sperm capacitation, 4D label-free quantitative proteomics was first applied to analyze the changes of sperm in Eriocheir sinensis under three sequential physiological conditions: seminal vesicles (X2), hatched with the seminal receptacle content (X3), and incubated with egg water (X5). In total, 1536 proteins were identified, among which 880 proteins were quantified, with 82 and 224 proteins significantly altered after incubation with the seminal receptacle contents and egg water. Most differentially expressed proteins were attributed to biological processes by Gene Ontology annotation analysis. As the fundamental bioenergetic metabolism of sperm, the oxidative phosphorylation, glycolysis, and the pentose phosphate pathway presented significant changes under the treatment of seminal receptacle contents, indicating intensive regulation for sperm in the seminal receptacle. Additionally, the seminal receptacle contents also significantly increased the oxidation level of sperm, whereas the enhancement of abundance in superoxide dismutase, peroxiredoxin 1, and glutathione S-transferase after incubation with egg water significantly improved the resistance against oxidation. These results provided a new perspective for reproduction studies in crustaceans.

The multi-omics analysis in the hepatopancreas of Eriocheir sinensis provides novel insights into the response mechanism of heat stress.

Under global warming, heat stress can induce the excessive production of reactive oxygen species, causing irreversible damage to aquatic animals. It is essential to predict potentially harmful impacts on aquatic organisms under heat stress. Eriocheir sinensis, a typical crustacean crab, is widely distributed in China, American and Europe. Parent E. sinensis need migrate to the estuaries to reproduce in winter, and temperature is a key environmental factor. Herein, we performed a comprehensive transcriptomic and proteomic analysis in the hepatopancreas of E. sinensis under heat stress (20 °C and 30 °C), focusing on heat shock protein family, antioxidant system, energy metabolism and immune defense. The results revealed that parent E. sinensis generated adaptative responses to maintain physiological function under 20 °C stress via the transcriptional up-regulation of energy metabolism enzymes, mRNA synthesis and heat shock proteins. The transcriptional inhibition of key enzymes related to energy metabolism implied that 30 °C stress may lead to the dysfunction of energy metabolism in parent E. sinensis. Meanwhile, parent E. sinensis also enhanced the expression of ferritin and phospholipase D at translational level, and the glutathione s-transferase and heat shock protein 70 at both transcriptional and translational levels, speculating that parent E. sinensis can strengthen antioxidant and immune capacity to resist oxidative stress under 30 °C stress. This study elucidated the potential molecular mechanism in response to heat stress of parent E. sinensis hepatopancreas. The preliminary selection of heat tolerance genes or proteins in E. sinensis can provide a reference for the population prediction and the study of evolutionary mechanism under heat stress in crabs.

The role of the antennal glands and gills in acid-base regulation and ammonia excretion of a marine osmoconforming brachyuran.

The excretory mechanisms of stenohaline marine osmoconforming crabs are often compared to those of the more extensively characterized euryhaline osmoregulating crabs. These comparisons may have limitations, given that unlike euryhaline brachyurans the gills of stenohaline marine osmoconformers possess ion-leaky paracellular pathways and lack the capacity to undergo ultrastructural changes that can promote ion-transport processes in dilute media. Furthermore, the antennal glands of stenohaline marine osmoconformers are poorly characterized making it difficult to determine what role urinary processes play in excretion. In the presented study, ammonia excretory processes as well as related acid-base equivalent transport rates and mechanisms were investigated in the Dungeness crab, Metacarcinus magister - an economically valuable stenohaline marine osmoconforming crab. Isolated and perfused gills were found to predominantly eliminate ammonia through a microtubule network-dependent active NH4+ transport mechanism that is likely performed by cells lining the arterial pockets of the gill lamella where critical Na+/K+-ATPase detection was observed. The V-type H+-ATPase - a vital component to transbranchial ammonia excretion mechanisms of euryhaline crabs - was not found to contribute significantly to ammonia excretion; however, this may be due to the transporter's unexpected apical localization. Although unconnected to ammonia excretion rates, a membrane-bound isoform of carbonic anhydrase was localized to the apical and basolateral membranes of lamella suited for respiration. Urine was found to contain significantly less ammonia as well as carbonate species than the hemolymph, indicating that unlike those of some euryhaline crabs the antennal glands of the Dungeness crab reabsorb these molecules rather than eliminate them for excretion.

Comparative transcriptomic characterization of the ovary in the spawning process of the mud crab Scylla paramamosain.

Oviposition is induced upon mating in most insects. Spawning is a physiological process that is fundamental for the reproduction of Scylla paramamosain. However, the molecular mechanisms underlying the spawning process in this species are poorly understood. Herein, comprehensive ovary transcriptomic analysis was conducted at the germinal vesicle breakdown stage (GVBD), spawning stage, 0.5 h post-spawning stage, and 24 h post-spawning stage of S. paramamosain for gene discovery. A total of 67,230 unigenes were generated, and 27,975 (41.61%) unigenes were annotated. Meanwhile, the differentially expressed genes (DEGs) between the different groups were identified, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis was subsequently conducted. These results suggested that octopamine (OA) and tyramine (TA) could induce oviposition, while dopamine (DA) and serotonin (5-hydroxytryptamine [5-HT]) inhibit oviposition. The 20-hydroxyecdysone (20E) and methyl farnesoate (MF) signal pathways might be positively associated with oviposition. Furthermore, numerous transcripts that encode neuropeptides and their G-protein-coupled receptors (GPCRs), such as CNMamide, RYamide, ecdysis-triggering hormone (ETH), GPA2/GPB5 receptor, and Moody receptor, appear to be differentially expressed during the spawning process. Eleven unigenes were selected for qRT-PCR and the pattern was found to be consistent with the transcriptome expression pattern. Our work is the first spawning-related investigation of S. paramamosain focusing on the ovary at the whole transcriptome level. These findings assist in improving our understanding of spawning regulation in S. paramamosain and provide information for oviposition studies in other crustaceans.

The effect of anthropogenic substrate-borne vibrations on locomotion of the fiddler crab Austruca lactea.

The anthropogenic construction activities on the coasts, such as pile-driving, generate vibrations that propagate through the substrate. Such substrate-borne vibrations could potentially affect marine organisms inhabiting the benthic environments. However, there is a lack of documented studies on the effects of vibrations on benthic animals. To investigate whether anthropogenic substrate-borne vibrations such as pile-driving operation influence the fiddler crab, Austruca lactea, we measured their locomotion response under vibrations of 35, 120, 250, 500, and 750 Hz generated by a vibrator. We compared the locomotion of crabs between control and vibration-treatment groups using videography. The duration of movements was significantly lower under 120 Hz vibrations compared to the control. Moreover, crab velocity was significantly higher under vibrations of 120 Hz and 250 Hz compared to the control group. Our result suggests that A. lactea can detect low-frequency substrate-borne vibrations and experience stress, leading to increased energy consumption.

Short-term exposure to high pCO2 leads to decreased branchial cytochrome C oxidase activity in the presence of octopamine in a decapod.

In a recent mechanistic study, octopamine was shown to promote proton transport over the branchial epithelium in green crabs, Carcinus maenas. Here, we follow up on this finding by investigating the involvement of octopamine in an environmental and physiological context that challenges acid-base homeostasis, the response to short-term high pCO2 exposure (400 Pa) in a brackish water environment. We show that hyperregulating green crabs experienced a respiratory acidosis as early as 6 h of exposure to hypercapnia, with a rise in hemolymph pCO2 accompanied by a simultaneous drop of hemolymph pH. The slightly delayed increase in hemolymph HCO3- observed after 24 h helped to restore hemolymph pH to initial values by 48 h. Circulating levels of the biogenic amine octopamine were significantly higher in short-term high pCO2 exposed crabs compared to control crabs after 48 h. Whole animal metabolic rates, intracellular levels of octopamine and cAMP, as well as branchial mitochondrial enzyme activities for complex I + III and citrate synthase were unchanged in posterior gill #7 after 48 h of hypercapnia. However, application of octopamine in gill respirometry experiments suppressed branchial metabolic rate in posterior gills of short-term high pCO2 exposed animals. Furthermore, branchial enzyme activity of cytochrome C oxidase decreased in high pCO2 exposed crabs after 48 h. Our results indicate that hyperregulating green crabs are capable of quickly counteracting a hypercapnia-induced respiratory acidosis. The role of octopamine in the acclimation of green crabs to short-term hypercapnia seems to entail the alteration of branchial metabolic pathways, possibly targeting mitochondrial cytochrome C in the gill. Our findings help advancing our current limited understanding of endocrine components in hypercapnia acclimation. SUMMARY STATEMENT: Acid-base compensation upon short-term high pCO2 exposure in hyperregulating green crabs started after 6 h and was accomplished by 48 h with the involvement of the biogenic amine octopamine, accumulation of hemolymph HCO3-, and regulation of mitochondrial complex IV (cytochrome C oxidase).

Metagenomic analysis of gut microbiome illuminates the mechanisms and evolution of lignocellulose degradation in mangrove herbivorous crabs.

BACKGROUND: Sesarmid crabs dominate mangrove habitats as the major primary consumers, which facilitates the trophic link and nutrient recycling in the ecosystem. Therefore, the adaptations and mechanisms of sesarmid crabs to herbivory are not only crucial to terrestrialization and its evolutionary success, but also to the healthy functioning of mangrove ecosystems. Although endogenous cellulase expressions were reported in crabs, it remains unknown if endogenous enzymes alone can complete the whole lignocellulolytic pathway, or if they also depend on the contribution from the intestinal microbiome. We attempt to investigate the role of gut symbiotic microbes of mangrove-feeding sesarmid crabs in plant digestion using a comparative metagenomic approach. RESULTS: Metagenomics analyses on 43 crab gut samples from 23 species of mangrove crabs with different dietary preferences revealed a wide coverage of 127 CAZy families and nine KOs targeting lignocellulose and their derivatives in all species analyzed, including predominantly carnivorous species, suggesting the crab gut microbiomes have lignocellulolytic capacity regardless of dietary preference. Microbial cellulase, hemicellulase and pectinase genes in herbivorous and detritivorous crabs were differentially more abundant when compared to omnivorous and carnivorous crabs, indicating the importance of gut symbionts in lignocellulose degradation and the enrichment of lignocellulolytic microbes in response to diet with higher lignocellulose content. Herbivorous and detritivorous crabs showed highly similar CAZyme composition despite dissimilarities in taxonomic profiles observed in both groups, suggesting a stronger selection force on gut microbiota by functional capacity than by taxonomy. The gut microbiota in herbivorous sesarmid crabs were also enriched with nitrogen reduction and fixation genes, implying possible roles of gut microbiota in supplementing nitrogen that is deficient in plant diet. CONCLUSIONS: Endosymbiotic microbes play an important role in lignocellulose degradation in most crab species. Their abundance is strongly correlated with dietary preference, and they are highly enriched in herbivorous sesarmids, thus enhancing their capacity in digesting mangrove leaves. Dietary preference is a stronger driver in determining the microbial CAZyme composition and taxonomic profile in the crab microbiome, resulting in functional redundancy of endosymbiotic microbes. Our results showed that crabs implement a mixed mode of digestion utilizing both endogenous and microbial enzymes in lignocellulose degradation, as observed in most of the more advanced herbivorous invertebrates.