Sea Cucumber Apostichopus Japonicus Research Articles

Incorporating physiological knowledge into correlative species distribution models minimizes bias introduced by the choice of calibration area

AbstractCorrelative species distribution models (SDMs) are important tools to estimate species’ geographic distribution across space and time, but their reliability heavily relies on the availability and quality of occurrence data. Estimations can be biased when occurrences do not fully represent the environmental requirement of a species. We tested to what extent species’ physiological knowledge might influence SDM estimations. Focusing on the Japanese sea cucumber Apostichopus japonicus within the coastal ocean of East Asia, we compiled a comprehensive dataset of occurrence records. We then explored the importance of incorporating physiological knowledge into SDMs by calibrating two types of correlative SDMs: a naïve model that solely depends on environmental correlates, and a physiologically informed model that further incorporates physiological information as priors. We further tested the models’ sensitivity to calibration area choices by fitting them with different buffered areas around known presences. Compared with naïve models, the physiologically informed models successfully captured the negative influence of high temperature on A. japonicus and were less sensitive to the choice of calibration area. The naïve models resulted in more optimistic prediction of the changes of potential distributions under climate change (i.e., larger range expansion and less contraction) than the physiologically informed models. Our findings highlight benefits from incorporating physiological information into correlative SDMs, namely mitigating the uncertainties associated with the choice of calibration area. Given these promising features, we encourage future SDM studies to consider species physiological information where available.

Effects of gut bacterial community on differential growth of sea cucumber (Apostichopus japonicus): Molecular mechanisms based on host-gut microbiome co-metabolism

Gut microbiome and their metabolites are increasingly recognized for their pivotal role in regulating the health and growth of the host. The mechanism by which the host-gut microbiome co-metabolism affects the growth rate of sea cucumber (Apostichopus japonicus) remains unclear. In this study, the intestinal wall tissues of fast-growing (FG) and slow-growing (SG) A. japonicus were collected from the full-sib family for 16S amplicon sequencing, metagenomics, metabolomics, and transcriptomics analyses. The results showed a marked variation in the gut bacterial community of A. japonicus with different growth rates. Compared with SG group, FG A. japonicus had a higher abundance of Bacteroidetes. Many of these bacteria were associated with the degradation of polysaccharides, which could potentially affect growth performance. The abundance of Proteobacteria, however, was found to be higher in SG A. japonicus, thereby increasing the potential risk of disease. The metabolome results were further supported by the predicated KEGG functions of the gut microbiomes, lots of metabolites related to the metabolism and biosynthesis of fructose 6-phosphate, arachidonic acid, oleic acid, and dihydroxyacetone phosphate were upregulated in the FG A. japonicus. Furthermore, the genes (IGF1, HK, PK, PCK1) were significantly upregulated in the FG A. japonicus, which were mainly enriched in the metabolic pathways of lipids, amino acids, and carbohydrates. Gut bacterial community via their unique metabolic pathways, which in turn affected the metabolic phenotypes of the organism and eventually led to differences in growth performance of A. japonicus. These findings might help understand the molecular mechanisms of growth gap in sea cucumber, thereby increasing the mariculture production.

Ferroptosis resists intracellular Vibrio splendidus AJ01 mediated by ferroportin in sea cucumber Apostichopus japonicus

Ferroptosis, a kind of programmed cell death, is characterized with iron-dependent lipid ROS buildup, which is considered as an important cellular immunity in resisting intracellular bacterial infection in mammalian macrophages. In this process, lipid ROS oxidizes the bacterial biofilm to inhibit intracellular bacteria. However, the function of ferroptosis in invertebrate remains unknown. In this study, the existence of ferroptosis in Apostichopus japonicus coelomocytes was confirmed, and its antibacterial mechanism was investigated. First, our results indicated that the expression of glutathione peroxidase (AjGPX4) was significantly inhibited by 0.21-fold (p < 0.01) after injecting A. japonicus with the ferroptosis inducer RSL3, and the contents of MDA (3.93-fold, p < 0.01), ferrous iron (1.40-fold, p < 0.01), and lipid ROS (3.10-fold, p < 0.01) were all significantly increased under this condition and simultaneously accompanied with mitochondrial contraction and disappearance of cristae, indicating the existence of ferroptosis in the coelomocytes of A. japonicus. Subsequently, the contents of ferrous iron (1.40-fold, p < 0.05), MDA (2.10-fold, p < 0.01), ROS (1.70-fold, p < 0.01), and lipid ROS (2.50-fold, p < 0.01) were all significantly increased, whereas the mitochondrial membrane potential and GSH/GSSG were markedly decreased by 0.68-fold (p < 0.05) and 0.69-fold (p < 0.01) under Vibrio splendidus (AJ01) infection. This process could be reversed by the iron-chelating agent deferoxamine mesylate, which indicated that AJ01 could induce coelomocytic ferroptosis. Moreover, the results demonstrated that the intracellular AJ01 load was clearly decreased to 0.49-fold (p < 0.05) and 0.06-fold (p < 0.01) after treating coelomocytes with RSL3 and ferrous iron, which indicated that enhanced ferroptosis could inhibit bacterial growth. Finally, subcellular localization demonstrated that ferrous iron efflux protein ferroportin (AjFPN) and intracellular AJ01 were co-localized in coelomocytes. After AjFPN interference (0.58-fold, p < 0.01), the signals of ferrous iron and lipid ROS levels in intracellular AJ01 were significantly reduced by 0.38-fold (p < 0.01) and 0.48-fold (p < 0.01), indicating that AjFPN was an important factor in the introduction of ferroptosis into intracellular bacteria. Overall, our findings indicated that ferroptosis could resist intracellular AJ01 infection via AjFPN. These findings provide a novel defense mechanism for aquatic animals against intracellular bacterial infection.

Importance of the ECM-receptor interaction for adaptive response to hypoxia based on integrated transcription and translation analysis.

Low dissolved oxygen (LO) conditions represent a major environmental challenge to marine life, especially benthic animals. For these organisms, drastic declines in oxygen availability (hypoxic events) can trigger mass mortality events and thus, act as agents of selection influencing the evolution of adaptations. In sea cucumbers, one of the most successful groups of benthic invertebrates, the exposure to hypoxic conditions triggers adaptive adjustments in metabolic rates and behaviour. It is unclear, however, how these adaptive responses are regulated and the genetic mechanisms underpinning them. Here, we addressed this knowledge gap by assessing the genetic regulation (transcription and translation) of hypoxia exposure in the sea cucumber Apostichopus japonicus. Transcriptional and translational gene expression profiles under short- and long-term exposure to low oxygen conditions are tightly associated with extracellular matrix (ECM)-receptor interaction in which laminin and collagen likely have important functions. Finding revealed that genes with a high translational efficiency (TE) had a relatively short upstream open reading frame (uORF) and a high uORF normalized minimal free energy, suggesting that sea cucumbers may respond to hypoxic stress via altered TE. These results provide valuable insights into the regulatory mechanisms that confer adaptive capacity to holothurians to survive oxygen deficiency conditions and may also be used to inform the development of strategies for mitigating the harmful effects of hypoxia on other marine invertebrates facing similar challenges.

Immune responses of sea cucumber (Apostichopus japonicus) to combined environmental stress from high temperature and oil pollution

Context High temperature and oil pollution are typical examples of environmental stress to sea cucumber (Apostichopus japonicus) in northern China. Aims Evaluate the combined effects of high temperature and oil pollution on the immune responses of sea cucumbers. Methods In this study, we investigated the immune responses in the respiratory tree of sea cucumber subjected to the optimal temperature (16°C), high temperature (26°C), Oman crude oil (OCO) water-accommodated fractions (WAF) at 16°C (WAF + 16°C) and OCO WAF at 26°C (WAF + 26°C) for 48 h. Key results All three treatments significantly up-regulated total antioxidant capacity and disturbed active oxygen species homeostasis in sea cucumbers. Moreover, all three treatments caused immune-enzyme activity disorders, manifested by a significant increase in acid–alkaline phosphatase and nitric oxide synthase activities and a noticeable decline in lysozyme activity. Conclusions Combining the integrated biomarker-response index (WAF + 26°C &gt; 26°C &gt; WAF + 16°C), the combined stress could result in a more adverse effect on the immune responses of sea cucumbers than do high temperature or oil pollution alone, which might further exacerbate the health challenges for sea cucumbers in response to environmental stress. Implications This study has provided an insight into the immune impacts in benthic organisms caused by high temperature and oil pollution.

A multi-objective optimization approach for green supply chain network design for the sea cucumber (Apostichopus japonicus) industry

In China, aquatic supply chain network design does not include the green concept or the coordination of environmental and economic performance. Sea cucumber (Apostichopus japonicus) is an aquatic product of high economic value; however, studies on sea cucumber supply chain network optimization are lacking. This study is the first to design the sea cucumber supply chain and construct an optimization model. Considering the characteristics of the sea cucumber industry, LCA for Experts software and the CML-IA-Aug. 2016-world method were used to assess each aquaculture model's global warming potential (GWP), as the environmental performance indicator. In addition, multi-objective genetic algorithm (MOGA) coupled with Modified Technique for Order of Preference by Similarity to Ideal Solution (M-TOPSIS) integrates yield production, economic benefits, and environmental performance. The results demonstrated that cage seed rearing (CSR) combined bottom sowing aquaculture (BSA) represents the best production strategy upstream of the sea cucumber supply chain. In the downstream, the best proportion of sales channels in supermarkets, boutique stores and online shops accounted for 14.79 %, 58.02 % and 27.19 % of the production, respectively. The proposed optimization scenario 4 (S4) can increase product profit by 27.88 % and reduce GWP by 56.89 %. The following improvement measures are proposed: using sea cucumber aquaculture industry standards (cleaner production and green supplier selection) to regulate the behavior of enterprises, adopting an ecological and green production strategy, eliminating high-energy consumption and high emission production practices, and promoting widespread adoption of green consumption concepts. Finally, these measures may improve the sea cucumber supply chain, achieve coordinated environmental and economic performance development in the sea cucumber industry, and provide guidance for green optimization of other aquatic product supply chains in China.

A Novel Frontier in the Geographic Distribution of the Japanese Sea Cucumber Apostichopus japonicus (Selenka, 1867) (Stichopodidae: Holothuroidea) in the World

The natural distribution of the Japanese sea cucumber (Apostichopus japonicus Selenka, 1867), a sea cucumber species, is characterized by the Northwest Pacific. Long-term monitoring studies from 2020 to 2024 document a significant deviation from the known natural distribution of A. japonicus and its ability to extensively colonize the Gulf of Izmit, the easternmost part of the Sea of Marmara. This record is also the first documentation of the species from the Mediterranean Sea. Population data extracted from 67 samples allowed the determination of size and weight distribution, length-weight relationship, and condition factor. The study delves into the implications for fisheries management by evaluating the species’ dispersion beyond its natural range and its adaptation, shedding light on potential threats from illegal hookah diving fisheries. While its presence offers potential economic benefits through fishing income, its emergence as an invasive species can pose critical ecological risks to the receiving ecosystem.

Triterpene Glycosides from the Viscera of Sea Cucumber Apostichopus japonicus with Embryotoxicity.

Sea cucumbers release chemical repellents from their guts when they are in danger from predators or a hostile environment. To investigate the chemical structure of the repellent, we collected and chemically analyzed the viscera of stressed sea cucumbers (Apostichopus japonicus) in the Yellow Sea of China. Two undescribed triterpene glycosides (1 and 2), together with a known cladoloside A (3), were identified and elucidated as 3β-O-{2-O-[β-d-quinovopyranosyl]-4-O-[3-O-methyl-β-d-glucopyranosyl-(1→3)-β-d-glucopyranosyl]-β-d-xylopyranosyl}-holosta-9(11),25(26)-dien-16-one (1), 3β-O-{2-O-[β-d-glucopyranosyl]-4-O-[3-O-methyl-β-d-glucopyranosyl-(1→3)-β-d-glucopyranosyl]-β-d-xylopyranosyl}-holosta-9(11),25(26)-dien-16-one (2), 3β-O-{2-O-[3-O-methyl-β-d-glucopyranosyl-(1→3)-β-d-xylopyranosyl-(1→4)-β-d-quinovopyranosyl]-β-d-xylopyranosyl}-holosta-9(11),25(26)-dien-16-one (3) by spectroscopic analysis, including HR-ESI-MS and NMR spectra. Compounds 1, 2, and 3 display embryonic toxicity, as indicated by their 96-hour post-fertilization lethal concentration (96 hpf-LC50) values of 0.289, 0.536, and 0.091 μM, respectively. Our study discovered a class of triterpene glycoside compounds consisting of an oligosaccharide with four sugar units and a holostane aglycone. These compounds possess embryotoxicity and may serve as chemical defense molecules in marine benthic ecosystems.

Investigation of the Presence of Fibrillin in Sea Cucumber (Apostichopus japonicus) Body Wall by Utilizing Targeted Proteomics and Visualization Strategies.

Fibrillin is an important structural protein in connective tissues. The presence of fibrillin in sea cucumber Apostichopus japonicus is still poorly understood, which limits our understanding of the role of fibrillin in the A. japonicus microstructure. The aim of this study was to clarify the presence of fibrillin in the sea cucumber A. japonicus body wall. Herein, the presence of fibrillin in sea cucumber A. japonicus was investigated by utilizing targeted proteomics and visualization strategies. The contents of three different isoforms of fibrillin with high abundance in A. japonicus were determined to be 0.96, 2.54, and 0.15 μg/g (wet base), respectively. The amino acid sequence of fibrillin (GeneBank number: PIK56741.1) that started at position 631 and ended at position 921 was selected for cloning and expressing antigen. An anti-A. japonicus fibrillin antibody with a titer greater than 1:64 000 was successfully obtained. It was observed that the distribution of fibrillin in the A. japonicus body wall was scattered and dispersed in the form of fibril bundles at the microscale. It further observed that fibrillin was present near collagen fibrils and some entangled outside the collagen fibrils at the nanoscale. Moreover, the stoichiometry of the most dominant collagen and fibrillin molecules in A. japonicus was determined to be approximately 250:1. These results contribute to an understanding of the role of fibrillin in the sea cucumber microstructure.

Transcriptome analysis of sea cucumber (Apostichopus japonicus) in southern China under heat stress

Sea cucumber (Apostichopus japonicus) is a significant aquatic economic species globally. However, the advancement of the sea cucumber aquaculture industry is impeded by its high susceptibility to elevated temperatures. To elucidate the impact of high temperature on physiological changes, gene expression, and metabolic responses in sea cucumbers, we conducted transcriptional profiling of A. japonicus under heat stress. Our results revealed 3691 differentially expressed genes (DEGs), comprising 1629 up-regulated and 2062 down-regulated genes, between the control and treatment groups. Analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology (GO), and Protein-Protein Interaction Networks (PPI) indicated that high-temperature stress triggered the expression of stress defense-related genes while inhibiting the expression of genes associated with development and metabolism. Furthermore, 30 out of 39 DEGs were annotated to heat shock protein genes, underscoring the crucial role of these genes in the regulatory mechanism of A. japonicus in response to heat stress. In summary, our findings provide fundamental insights for further investigation into the molecular mechanisms in A. japonicus under high-temperature stress and may contribute to the development of novel strategies for identifying heat-tolerant candidate genes through the analysis of their transcriptional changes in the future.

Evaluation of the chronic toxicity of bisphenol A and bisphenol AF to sea cucumber Apostichopus japonicus after long-term single and combined exposure at environmental relevant concentration

Bisphenols are emerging endocrine disrupting pollutant, and several studies have reported that they are already ubiquitous in various environmental matrices and intend to deposit in sediment. The primary sources of bisphenols are river and sewage discharge. Sea cucumber (Apostichopus japonicus), a typical deposit feeder, is one of the most important commercial marine species in Aisa. However, the effects of the bisphenol A (BPA) and its analogues bisphenol AF (BPAF) on sea cucumber was unclear. In this study, we carried out field survey in major sea cucumber farming areas in northern China, with the aim of determining which bisphenol analogue is the major bisphenol contamination in this aquaculture area. The results showed that the presence of BPAF was detected in four sampling sites (Dalian, Tangshan, Laizhou, and Longpan). The mean level of BPAF in Laizhou sediment samples was the highest which reached to 9.007 ± 4.702 μ g/kg. Among the seawater samples, the BPAF only have been detected in the samples collected at Longpan. (0.011 ± 0.003 μ g/L). Furthermore, we conducted an experiment to evaluate the single and combined toxicity of BPA and BPAF on sea cucumbers. The concentrations were informed by the findings based on the results of field research. (0.1, 1.0, and 10 μ g/L). After exposure, the body weight gain, and specific growth rate showed no significant changes (P > 0.05). We observed the histological alterations in respiratory tree of treated sea cucumbers including the fusion and detachment of lining epithelial tissue, and increase of lumen space. However, the catalase (CAT), malondialdehyde (MDA), and glutathione (GSH) activity was not significantly changed (P > 0.05). We evaluated the effects of BPA and BPAF through calculating the integrated biomarker response index (IBR), and the results indicated that the toxicity of combined treatment was higher than single treatment. Additionally, BPAF exposure to A. japonicus was more toxic than BPA.

A2M possesses anti-bacterial functions by recruiting and enhancing phagocytosis through GRP78 in an echinoderm

Alpha-2-macroglobulin (A2M) is an extracellular macromolecule mainly known for its role as a broad-spectrum protease inhibitor in mammals. However, the immune recognition and regulation mechanisms of A2M in invertebrates are still not well investigated. In the current study, the role of sea cucumber Apostichopus japonicus A2M in the regulation of innate immune responses was explored. We found that AjA2M promotes phagocytosis of Vibrio splendidus in coelomocytes of sea cucumber. Then two major functional structural domains of AjA2M, the thioester domain (TED) and the receptor-binding structural domain (RBD) were cloned. It was found that the AjA2M-TED binds to pathogens while causing Vibrio splendidus aggregation; the AjA2M-RBD interacts with the Glucose Regulated Protein 78 (AjGRP78), subsequently AjGRP78 accelerates the degradation of Vibrio splendidus in lysosomes by facilitating polymerisation and rearrangement of the cytoskeleton. Collectively, the findings together suggest that A2M-GRP78 axis mediates immune signaling pathway of phagocytosis and AjA2M has been characterized to play an essential crucial role in antibacterial immune responses of invertebrates.

Integrated metabolome–metagenome analysis provides new insights into the impacts of sea cucumber (Apostichopus japonicus) aestivation on marine sediment composition

To explore the impacts of sea cucumber aestivation on sediment composition, sediments were sampled from aestivation sites and non-aestivation sites of a commercial Apostichopus japonicus culture pond, and metabolome–metagenome integrated analysis was performed. The results showed that 1) there was no significant difference in the microbial composition of sediments between aestivation and non-aestivation sites, but the relative abundances of some bacterial species changed slightly (especially pathogenic and spoilage species); 2) altered average relative abundances of potential host bacteria of antibiotic resistance genes (ARGs) were observed; 3) compared with non-aestivation site sediments, decreased average absolute abundances of two ARGs (tetW and floR) and increased average absolute abundance of one ARG (tetM) were observed in aestivation site sediments; and 4) a total of 74 significantly differentially expressed metabolites (SDMs) were identified in aestivation sites compared with non-aestivation sites. Four SDMs were closely correlated with antibacterial activity and bacterial physiology. All data observed in this study indicate that sea cucumber aestivation does not affect sediment microbial composition but does influence the relative abundances of some bacteria species; in addition, tetM might be considered a key indicator for monitoring and indicating ARG transfer or the sediment status of sea cucumber culture ponds during aestivation. In conclusion, the observations of this study enrich our knowledge of the impacts of the physiological behavior of marine ecosystem inhabitants on their ecosystem and provide novel insights for the management of aquaculture activities.

Sea Cucumber Viscera Contains Novel Non-Holostane-Type Glycoside Toxins that Possess a Putative Chemical Defense Function.

Sea cucumbers frequently expel their guts in response to predators and an aversive environment, a behavior perceived as releasing repellents involved in chemical defense mechanisms. To investigate the chemical nature of the repellent, the viscera of stressed sea cucumbers (Apostichopus japonicus) in the Yellow Sea of China were collected and chemically analyzed. Two novel non-holostane triterpene glycosides were isolated, and the chemical structures were elucidated as 3ꞵ-O-[ꞵ-D-glucopyranosyl-(1→2)-ꞵ-D-xylopyranosyl]-(20S)-hydroxylanosta-7,25-diene-18(16)-lactone (1) and 3ꞵ-O-[ꞵ-D-quinovopyranosyl-(1→2)-ꞵ-D-xylopyranosyl]-(20S)-hydroxylanosta-7,25-diene-18(16)-lactone (2) by spectroscopic and mass-spectrometric analyses, exemplifying a triterpene glycoside constituent of an oligosaccharide containing two sugar-units and a non-holostane aglycone. Zebrafish embryos were exposed to various doses of 1 and 2 from 4 to 96 hpf. Compound 1 exposure showed 96h-LC50 41.5 µM and an increased zebrafish mortality rates in roughly in a dose- and time-dependent manner. Compound 2, with different sugar substitution, exhibited no mortality and moderate teratogenic toxicity with a 96h-EC50 of 173.5 µM. Zebrafish embryos exhibited teratogenic effects, such as reduced hatchability and total body length. The study found that triterpene saponin from A. japonicus viscera had acute toxicity in zebrafish embryos, indicating a potential chemical defense role in the marine ecosystem.

YTHDC1/CRM1 Facilitates m6A-Modified circRNA388 Nuclear Export to Induce Coelomocyte Autophagy via the miR-2008/ULK Axis in Apostichopus japonicus.

N 6-methyladenosine (m6A), the most prevalent internal modification in eukaryotic RNA, was able to mediate circular RNA (circRNA) function in many immune processes. Nevertheless, the functional role of m6A-modified circRNAs in innate immunity of invertebrates remained unclear. In this study, we identified m6A-modified circRNA388 from cultured sea cucumber (Apostichopus japonicus) coelomocytes, which was mainly detected in cytoplasm after Vibrio splendidus infection. A knockdown assay indicated that cytoplasm circRNA388 promoted coelomocyte autophagy and decreased the number of intracellular V. splendidus. Mechanistically, the circRNA388 in the cytoplasm directly sponged miR-2008 to block its interaction with Unc-51-like kinase 1 from A. japonicus (AjULK) and further promoted autophagy to resist V. splendidus infection. More importantly, we found that m6A modification was vital to circRNA388 nuclear export with YTH domain-containing protein 1 from A. japonicus (AjYTHDC1) as the reader. AjYTHDC1 facilitated the nuclear export of m6A-modified circRNA388 via interaction with exportin-1 (chromosomal maintenance 1) from A. japonicus (AjCRM1). Knockdown of AjCRM1 could significantly decrease the content of cytoplasm circRNA388. Overall, our results provide the first evidence that nuclear export of m6A-modified circRNA388 is dependent on the novel AjCRM1 to our knowledge, which was further promoted coelomocyte autophagy by miR-2008/AjULK axis to clear intracellular V. splendidus.

Effect of probiotic feed supplementation on skin ulceration syndrome in Apostichopus japonicus

Skin ulceration syndrome is among the most harmful diseases in sea cucumber (Apostichopus japonicus). The annual loss due to this disease is more than 1 billion dollars, which seriously affected the healthy and sustainable development of A. japonicus. In this paper, animals were divided into 4 experimental groups: a control group given basic feed, experimental groups with 1% Lactobacillus, 1% Bacillus, 1% mixture with 0.5% Lactobacillus and 0.5% Bacillus added to the basic feed. After feeding for 60 days, the samples were taken to analyze the growth performance, digestive enzyme activity, and immune indexes of A. japonicus. Furthermore, the resistance to the skin ulceration syndrome of A. japonicus was analyzed. The results showed that the probiotics promoted the growth performance of A. japonicus, the weight gain rate of the control group was 120.08±12.07%, the weight gain rate of the Lactobacillus group was 152.11±10.06%, the weight gain rate of the Bacillus group was 148.03±9.88%, and the weight gain rate of the mixed bacteria group was as high as 210.11±19.41%. The digestive enzyme activities in the intestinal tract of A. japonicus increased significantly in the probiotic-added groups, and the mixed bacteria were more effective in enhancing the digestive enzyme activities than single Bacillus and Lactobacillus. In addition, the probiotics could significantly increase the phagocytosis rate of the body cavity cells of A. japonicus. The phagocytic rate of Lactobacillus and Bacillus subtilis was comparable; the mixed probiotics were significantly higher than those of single probiotics, and this phagocytic rate gradually increased over time. After Vibrio splendidus and Pseudoalteromonas infection, the mortality rate of A. japonicus in the probiotic-added group was reduced, which indicated that probiotics could improve the immunity of A. japonicus, and the ability to resist the infection of Vibrio splendidus and Pseudoalteromonas was enhanced. Bacillus exhibits superior antimicrobial ability compared to Lactobacillus, and the mixed strain displays the most effective antimicrobial properties. This study provided a scientific basis for effectively preventing and controlling skin ulceration syndrome in the aquaculture of A. japonicus.

A Comparative Study on Effects of Three Butyric Acid-Producing Additives on the Growth Performance, Non-specific Immunity, and Intestinal Microbiota of the Sea Cucumber Apostichopus japonicus.

The providers of butyric acid, Clostridium butyricum (CB), sodium butyrate (SB), and tributyrin (TB), have been extensively studied as aquafeed additives in recent years. However, no comparative study has been reported on the probiotic effects of CB, SB, and TB as feed additives on sea cucumber (Apostichopus japonicus). A 63-day feeding trial was performed to assess the effects of dietary live cells of C. butyricum (CB group, the basal diet supplemented with 1% CB), sodium butyrate (SB group, the basal diet supplemented with 1% SB), and tributyrin (TB group, the basal diet supplemented with 1% TB) on the growth, non-specific immunity, and intestinal microbiota of A. japonicus with a basal diet group as the control. Results indicated that all three additives considerably increased A. japonicus growth, with dietary CB having the optimal growth-promoting effect. Of the seven non-specific enzyme parameters measured in coelomocytes of A. japonicus (i.e., the activities of phagocytosis, respiratory burst, superoxide dismutase, alkaline phosphatase, acid phosphatase, catalase, and lysozyme), dietary CB, SB, and TB considerably increased the activities of six, five, and six of them, respectively. The immune genes (Aj-p105, Aj-p50, Aj-rel, and Aj-lys) expression in the mid-intestine tissues of A. japonicus was significantly increased by all three additives. The CB group had the highest expression of all four genes. Additionally, the relative expression of Aj-p105, Aj-p50, and Aj-lys genes was significantly up-regulated in the three additive groups after stimulation with inactivated Vibrio splendidus. Dietary CB enhanced the intestinal microbial diversity and richness in A. japonicus while dietary TB decreased them. Meanwhile, dietary CB, SB, and TB significantly enhanced the abundance of Firmicutes, unclassified_f_Rhodobacteraceae, and Proteobacteria, respectively, while dietary CB and SB reduced the abundance of Vibrio. Dietary CB and SB improved the stability of microbial ecosystem in the intestine of A. japonicus. In contrast, dietary TB appeared to have a negative effect on the stability of intestinal microbial ecosystem. All three additives improved the intestinal microbial functions associated with energy production and immunity regulation pathways, which may contribute directly to growth promotion and non-specific immunity enhancement in A. japonicus. Collectively, in terms of enhancing growth and non-specific immunity, as well as improving intestinal microbiota, dietary live cells of C. butyricum exhibited the most effective effects in A. japonicus.

Targeted Affinity Purification and Mechanism of Action of Angiotensin-Converting Enzyme (ACE) Inhibitory Peptides from Sea Cucumber Gonads.

Protein hydrolysates from sea cucumber (Apostichopus japonicus) gonads are rich in active materials with remarkable angiotensin-converting enzyme (ACE) inhibitory activity. Alcalase was used to hydrolyze sea cucumber gonads, and the hydrolysate was separated by the ultrafiltration membrane to produce a low-molecular-weight peptide component (less than 3 kDa) with good ACE inhibitory activity. The peptide component (less than 3 kDa) was isolated and purified using a combination method of ACE gel affinity chromatography and reverse high-performance liquid chromatography. The purified fractions were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and the resulting products were filtered using structure-based virtual screening (SBVS) to obtain 20 peptides. Of those, three noncompetitive inhibitory peptides (DDQIHIF with an IC50 value of 333.5 μmol·L-1, HDWWKER with an IC50 value of 583.6 μmol·L-1, and THDWWKER with an IC50 value of 1291.8 μmol·L-1) were further investigated based on their favorable pharmacochemical properties and ACE inhibitory activity. Molecular docking studies indicated that the three peptides were entirely enclosed within the ACE protein cavity, improving the overall stability of the complex through interaction forces with the ACE active site. The total free binding energies (ΔGtotal) for DDQIHIF, HDWWKER, and THDWWKER were -21.9 Kcal·mol-1, -71.6 Kcal·mol-1, and -69.1 Kcal·mol-1, respectively. Furthermore, a short-term assay of antihypertensive activity in spontaneously hypertensive rats (SHRs) revealed that HDWWKER could significantly decrease the systolic blood pressure (SBP) of SHRs after intravenous administration. The results showed that based on the better antihypertensive activity of the peptide in SHRs, the feasibility of targeted affinity purification and computer-aided drug discovery (CADD) for the efficient screening and preparation of ACE inhibitory peptide was verified, which provided a new idea of modern drug development method for clinical use.

Transcriptome analysis of purple and green Apostichopus japonicus reared under different breeding environments

Sea cucumber (Apostichopus japonicus) is one of the cultured species with the highest production value to a single species in China’s marine agricultural industry with high nutritional and medicinal value. Body color influences numerous biological functions in sea cucumbers, and it has become a key trait in selective breeding. It is found that a cultivation environment with strong light intensity leads to a lighter body color of A. japonicus compared to dark conditions. However, little research has been conducted on the molecular mechanisms regulating body color in different breeding environments. In this study, we used Illumina sequencing to examine gene expression patterns in green and purple A. japonicus exposed to different culture conditions. Overall, the genes in the body wall of purple individuals were more affected by the environment. We also identified pathways that were potentially influenced by the breeding environment, such as “Drug metabolism - cytochrome P450”, “Porphyrin and chlorophyll metabolism”, “Phosphatidylinositol signaling system”, “TGF-beta signaling”, and “mTOR signaling”. The aim of this study was to determine the light conditions and breeding environment that are more favorable to the requirements of body color in A. japonicus and apply them to the selection of high-quality breeds. The results obtained will support the breeding of A. japonicus with specific traits and the production of seedlings to promote the development of this maricultural industry.

Dietary tryptophan decreases the impacts of seawater temperature changes on behavior and gut health of the sea cucumber Apostichopus japonicus

Seawater temperature change is an important concern for seed production and pond culture of sea cucumbers. The present study found that tentacle activity frequency was significantly lower in sea cucumbers exposed to continuous and rapid temperature increases than that of those at ambient temperature. Feeding behavior directly determines food intake, and further affects physiology and growth efficiency of sea cucumbers. This means that the decline in feeding caused by continuous and rapid temperature increases needs to be addressed in sea cucumber aquaculture. However, a sudden temperature change of 5 °C had no significant effect on behaviors of sea cucumbers. This indicates that continuous temperature increases, rather than a sudden increase, result in behavioral impacts on sea cucumbers. Therefore, we recommend aqua-farmers reduce the feeding amount for sea cucumbers during continuous and rapid temperature increases.In the present study, feeding behavior was significantly higher in sea cucumbers fed with 3% dietary tryptophan than that of those fed with 0% and 5% dietary tryptophan. This indicates that 3% dietary tryptophan increases the food intake of sea cucumbers, and mitigates the feeding decline caused by continuous and rapid temperature increase. This indicates that tryptophan has the potential to promote the feeding of sea cucumbers in seed production and pond culture. Adhesion capacity of sea cucumbers fed with 5% dietary tryptophan was significantly higher than that of individuals fed with 0% and 3% dietary tryptophan. This suggests that dietary tryptophan increases the feeding of sea cucumbers exposed to continuous and rapid temperature increases in pond culture and seed production. In addition, this study found that sea cucumbers fed with 3% dietary tryptophan had higher intestinal colony richness under the continuously rapid temperature change. The present study provides an effective method to improve adhesion behavior and to alleviate the impacts on feeding behavior for seed production and pond culture of sea cucumbers exposed to continuous and rapid temperature increases.