Abalone Haliotis Discus Hannai Research Articles

Transcriptome analysis of hybrid abalone (Haliotis discus hannai ♀ × H. fulgens ♂) reveals non-additive effects contributing to growth heterosis at early summer water temperature in Fujian

Transcriptome analysis of hybrid abalone (Haliotis discus hannai ♀ × H. fulgens ♂) reveals non-additive effects contributing to growth heterosis at early summer water temperature in Fujian

Allatotropin (AT) related peptides L-ATRP and D2-ATRP diastereomers activate an endogenous receptor and suppress heart rate in the Pacific abalone Haliotis discus hannai

Allatotropin (AT) has been identified in many insects and plays important roles in the regulation of their intestinal contraction, heart rate, ion transport, and digestive enzyme secretion. However, information on AT-related bioinformatics in other animal phyla is scarce. In this study, we cloned a full-length cDNA encoding the AT-related peptide receptor (ATRPR) of the abalone Haliotis discus hannai (Hdh) and further characterized Hdh-ATRPR with its potential ligands, Hdh-ATRPs. In luciferase reporter and Ca2+ mobilization assays, Hdh-ATRPs, including a D-type Phe at the second amino acid position, Hdh-D2-ATRP, activated Hdh-ATRPR in a dose-dependent manner, whereas all-L-type Hdh-ATRP was a more potent ligand than Hdh-D2-ATRP. Furthermore, Hdh-ATRPs induced ERK1/2 phosphorylation in Hdh-ATRPR-expressing HEK293 cells, which was dose-dependently abolished by the PKC inhibitor Gö6983. The heart rate decreased significantly within 10 min when Hdh-D2-ATRP was injected into the adduct muscle sinus of abalone (0.2 or 1.0 µg/g body weight), while the abalone injected with a high concentration of Hdh-D2-ATRP (1.5 μg/g body weight) were sublethal within 5 h. Thus, Hdh-ATRP signaling is primarily linked to the Gαq/PKC and is possibly associated with heart rate regulation in abalone.

Fasting time after feeding affects the evaluation of metabolic response in abalone Haliotis discus hannai to nutrients using dietary methionine as an example

As one of the essential amino acid, methionine (Met) is the initiating amino acid for proteins synthesis, and it can also act as a molecular signal regulating various metabolic processes. Meanwhile, the postprandial metabolic responses varied greatly among different species, nutrients and fasting time. In the present study, postprandial metabolic responses of abalone Haliotis discus hannai fed with different levels of dietary methionine (0.46%, 1.19% and 2.07%, respectively) for 84 days were investigated. Compared with the group with 0.46% of dietary methionine, 2.07% of dietary methionine significantly increased the concentration of glucose in CFH at 0 h, 12 h, 24 h, 36 h, 60 h and 84 h, while the concentrations of ammonia were significantly decreased by 2.07% of dietary methionine at each time point after feeding (P < 0.05). The 1.19% of dietary methionine significantly increased the activity of trypsin at 0 h, 24 h 60 h and 84 h after feeding (P < 0.05). The phosphorylation levels of protein kinase B (AKT), mechanistic target of rapamycin (mTOR) and ribosomal s6 protein (S6) peaked at 12 h and then dropped to base level at 36–84 h. Compared with the group with 0.46% of dietary methionine, 1.19% of dietary methionine significantly increased the phosphorylation levels of AKT at 12 h and 24 h, and extended the phosphorylation time of AKT (P < 0.05). And the phosphorylation level of mTOR was significantly increased by 1.19% of dietary methionine at 12 h, 24 h and 36 h after feeding (P < 0.05). Significant activation or repression of the mRNA levels related to glucose and lipid metabolism were observed at 12–24 h after feeding, and most of which were remain stable at 36–84 h (P < 0.05). The expressions of genes related of insulin signaling pathway (phosphoinositide 3-kinase and protein kinase B), glycolysis (hexokinase) and transport of lipid (fatty acid transport protein) were significantly increased by 1.19% of dietary methionine, while the key gene of gluconeogenesis (glucose-6-phosphatase) were decreased. In summary, 1.19% of dietary methionine could enhance digestive and absorptive capacity, protein synthesis, glucose metabolism and fatty acid transportation of abalone. Take long-term feeding trial with dietary methionine as an example and combine the results of data analysis, it is suggested to analyze the protein level of mTOR signaling pathway within 36 h after feeding, assess the biochemical parameters in CFH and the mRNA levels of nutrient metabolism related genes within 36–84 h, and detect the activities of digestive and absorptive enzymes within 60–84 h.

Structure and function of vasa gene in gonadal gametogenesis of Pacific abalone

Pacific abalone (Haliotis discus hannai) is a marine gastropod mollusc with significant economic importance in both global fisheries and aquaculture. However, studies exploring the gonadal development and regulatory mechanisms of Haliotis discus hannai are limited. This study aimed to explore whether the vasa gene acted as a molecular marker for germ cells. Initially, the vasa gene was successfully cloned using the cDNA-end rapid amplification technique. The cloned gene had a 2478-bp-long open reading frame and encoded 825 amino acids. Then, a recombinant expression vector was constructed based on the Vasa protein, and an 87-kDa recombinant protein was prepared. Subsequently, a polyclonal antibody was prepared using the purified recombinant protein. The enzyme-linked immunosorbent assay (ELISA) confirmed the titer of the antibody to be ≥512 K. The immunohistochemical analysis revealed that Vasa was widely expressed in oogonia, Stage I oocytes, spermatogonia, and primary spermatocytes. The specific expression of Vasa in the hermaphroditic gonads of abalone was assessed using western blotting to investigate the effects of different photoperiods (12 L:12D, 24 L:0D, 18 L:6D, and 6 L:18D) on the gonadal development of abalone (P < 0.05), with higher expression levels observed in the ovarian proliferative and spermary maturing stages compared with other developmental stages (P < 0.05). Additionally, Vasa exhibited the highest expression in the spermary and ovary under a photoperiod of 18 L:6D (P < 0.05). These data demonstrated the key role of Vasa in developing germ cells in abalone. They shed light upon the molecular mechanism through which the photoperiod influenced Vasa expression and regulated gonadal development in abalone. The findings might provide theoretical references for analyzing the differentiation pattern of abalone germ cells and the genetic improvement and conservation of germplasm resources.

Carryover effects of embryonic hypoxia exposure on adult fitness of the Pacific abalone

The widespread and severe drop in dissolved oxygen concentration in the open ocean and coastal waters has attracted much attention, but assessments of the impacts of environmental hypoxia on aquatic organisms have focused primarily on responses to current exposure. Past stress exposure might also affect the performance of aquatic organisms through carryover effects, and whether these effects scale from positive to negative based on exposure degree is unknown. We investigated the carryover effects of varying embryonic hypoxia levels (mediate hypoxia: 3.0−3.1 mg O2/L; severe hypoxia: 2.0−2.1 mg O2/L) on the fitness traits of adult Pacific abalone (Haliotis discus hannai), including growth, hypoxia tolerance, oxygen consumption, ammonia excretion rate, and biochemical responses to acute hypoxia. Moderate embryonic hypoxia exposure significantly improved the hypoxia tolerance of adult Pacific abalone without sacrificing growth and survival. Adult abalone exposed to embryonic hypoxia exhibited physiological plasticity, including decreased oxygen consumption rates under environmental stress, increased basal methylation levels, and a more active response to acute hypoxia, which might support their higher hypoxia tolerance. Thus, moderate oxygen declines in early life have persistent effects on the fitness of abalone even two years later, further affecting population dynamics. The results suggested that incorporating the carryover effects of embryonic hypoxia exposure into genetic breeding programs would be an important step toward rapidly improving the hypoxia tolerance of aquatic animals. The study also inspires the protection of endangered wild animals and other vulnerable species under global climate change.

Shellfish CO2 excretion is modulated by seawater carbonate chemistry but largely independent of pCO2.

Four species of shellfish, blue mussel (Mytilus galloprovincialis), Pacific abalone (Haliotis discus hannai), zhikong scallops (Chlamys farreri), and Pacific oyster (Crassostrea gigas), were exposed to decoupled carbonate system variables to investigate the impacts of different seawater carbonate parameters on the CO2 excretion process of mariculture shellfish. Six experimental groups with two levels of seawater pH (pH 8.1 and pH 7.7) and three levels of total alkalinity (TA = 1000, 2300, and 3600μmol/kg, respectively) were established, while pH 8.1 and TA = 2300μmol/kg was taken as control. Results showed that the CO2 excretion rates of these tested shellfish were significantly affected by the change in carbonate chemistry (P < 0.05). At the same TA level, animals incubated in the acidified group (pH 7.7) had a lower CO2 excretion rate than those in the control group (pH 8.1). In comparison, at the same pH level, the CO2 excretion rate increased when seawater TA level was elevated. No significant correlation between the CO2 excretion rate and seawater pCO2 levels (P > 0.05) was found; however, a significant correlation (P < 0.05) between CO2 excretion rate and TA-DIC (the difference between total alkalinity and dissolved inorganic carbon) was observed. Blue mussel has a significantly higher CO2 excretion rate than the other three species in the CO2 excretions per unit mass of soft parts, with no significant difference observed among these three species. However, in terms of CO2 excretion rate per unit mass of gills, abalone has the highest CO2 excretion rate, while significant differences were found between each species. Our studies indicate that the CO2 buffering capacity impacts the CO2 excretion rate of four shellfish species largely independent of pCO2. Since CO2 excretion is related to acid-base balancing, the results imply that the effects of other carbonate parameters, particularly the CO2 buffering capacity, should be studied to fully understand the mechanism of how acidification affects shellfish. Besides, the species difference in gill to soft parts proportion may contribute to the species difference in responding to ocean acidification.

Genetic diversity and population structure of Pacific abalone (Haliotis discus hannai) using SNP genotyping data

The Pacific abalone Haliotis discus hannai is one of the most valuable mollusks in China. It also supports an important fishery in the northern coast of China, which has shown a declining tendency. The conservation of this species is attracting attentions under the potential threats of alien stocks and hybrid varieties disseminated in aquaculture. However, population genetics information on whole-genome level is lacking. The present study analyzed the genetic diversity, population structure, and inbreeding coefficient of nine populations collected from China and Japan using single nucleotide polymorphism (SNP) genotyping data from 214 samples. The results showed higher observed heterozygosity (Ho) than expected heterozygosity (He), and negative inbreeding coefficient (Fis) values indicating an excess of heterozygotes, and highlighting medium to high genetic diversity within the population. The genetic structures of the nine populations were categorized into three distinct subgroups. Pairwise Fst values based on all SNP loci and those for neutral markers were consistently low, indicating an overall genetic homogeneity between the populations. The distribution of runs of homozygosity (ROH) lengths suggests the potential occurrence of kinship relationships in various historical periods, including both distant and recent periods. The TreeMix and shared haplotypes suggested strong gene flow from Japanese populations of Pacific abalone to Chinese populations. These findings establish the stock status of the Pacific abalone in China, providing a basis for conservation, fishery management and utilization of the stocks in aquaculture.

Physicochemical and Rheological Properties of Degraded Konjac Gum by Abalone (Haliotis discus hannai) Viscera Enzyme.

In the present study, a new degraded konjac glucomannan (DKGM) was prepared using a crude enzyme from abalone (Haliotis discus hannai) viscera, and its physicochemical properties were investigated. After enzymatic hydrolysis, the viscosity of KGM obviously decreased from 15,500 mPa·s to 398 mPa·s. The rheological properties analysis of KGM and DKGMs revealed that they were pseudoplastic fluids, and pseudoplasticity, viscoelasticity, melting temperature, and gelling temperature significantly decreased after enzymatic hydrolysis, especially for KGM-180 and KGM-240. In addition, the molecular weight of KGM decreased from 1.80 × 106 Da, to 0.45 × 106 Da and the polydispersity index increased from 1.17 to 1.83 after 240 min of degradation time. Compared with natural KGM, the smaller particle size distribution of DKGM further suggests enzyme hydrolysis reduces the aggregation of molecular chains with low molecular weight. FT-IR and FESEM analyses showed that the fragmented KMG chain did not affect the structural characteristics of molecular monomers; however, the dense three-dimensional network microstructure formed by intermolecular interaction changed to fragment microstructure after enzyme hydrolysis. These results revealed that the viscosity and rheological properties of KGM could be controlled and effectively changed using crude enzymes from abalone viscera. This work provides theoretical guidance for the promising application of DKGM in the food industry.

First insight on the genetic base of the heritable variation of Pacific abalone (Haliotis discus hannai) resistance to Polydora hoplura infestation

Polydora hoplura is an invasive species and a harmful pest in commercial shellfish aquaculture, particularly for farmed abalone species. Different lines of treatment for Polydora affecting abalone have been explored, but with limited success. Using selective breeding to increase abalone resistance to and reduce shell damage from Polydora infestation could be a promising alternative. However, in abalone there is currently no information on the genetic variability for resistance to Polydora infestation. In the present study, heritability (h2) of traits associated with Pacific abalone resistance to P. hoplura were estimated after challenging abalone juveniles with trojan abalones heavily infested, using a nested experimental design, with 1 male crossed with 2–4 females. The heritability values were low but significant for some of those traits: survival h2=0.09; Infestation Intensity at Q50h2 = 0.16 ± 0.11; and the proportion of the shell area occupied by blisters caused by the mud-worms h2 = 0.29 ± 0.14. These results suggest a significant potential for improving resistance or tolerance to P. hoplura infestation through selective breeding in Pacific abalone.

Morphologic, cytometric, quantitative transcriptomic and functional characterisation provide insights into the haemocyte immune responses of Pacific abalone (Haliotis discus hannai).

In recent years, the abalone aquaculture industry has been threatened by the bacterial pathogens. The immune responses mechanisms underlying the phagocytosis of haemocytes remain unclear in Haliotis discus hannai. It is necessary to investigate the immune mechanism in response to these bacterial pathogens challenges. In this study, the phagocytic activities of haemocytes in H. discus hannai were examined by flow cytometry combined with electron microscopy and transcriptomic analyses. The results of Vibrio parahaemolyticus, Vibrio alginolyticus and Staphylococcus aureu challenge using electron microscopy showed a process during phagosome formation in haemocytes. The phagocytic rate (PP) of S. aureus was higher than the other five foreign particles, which was about 63%. The PP of Vibrio harveyi was about 43%, the PP peak of V. alginolyticus in haemocyte was 63.7% at 1.5h. After V. parahaemolyticus and V. alginolyticus challenge, acid phosphatase, alkaline phosphatase, total superoxide dismutase, lysozyme, total antioxidant capacity, catalase, nitric oxide synthase and glutathione peroxidase activities in haemocytes were measured at different times, differentially expressed genes (DEGs) were identified by quantitative transcriptomic analysis. The identified DEGs after V. parahaemolyticus challenge included haemagglutinin/amebocyte aggregation factor-like, supervillin-like isoform X4, calmodulin-like and kyphoscoliosis peptidase-like; the identified DEGs after V. alginolyticus challenge included interleukin-6 receptor subunit beta-like, protein turtle homolog B-like, rho GTPase-activating protein 6-like isoform X2, leukocyte surface antigen CD53-like, calponin-1-like, calmodulin-like, troponin C, troponin I-like isoform X4, troponin T-like isoform X18, tumor necrosis factor ligand superfamily member 10-like, rho-related protein racA-like and haemagglutinin/amebocyte aggregation factor-like. Some immune-related KEGG pathways were significantly up-regulated or down-regulated after challenge, including thyroid hormone synthesis, Th17 cell differentiation signalling pathway, focal adhesion, melanogenesis, leukocyte transendothelial migration, inflammatory mediator regulation of TRP channels, ras signalling pathway, rap1 signalling pathway. This study is the first step towards understanding the H. discus hannai immune system by adapting several tools to gastropods and providing a first detailed morpho-functional study of their haemocytes.

Dietary Clostridium autoethanogenum protein has dose-dependent influence on the gut microbiota, immunity, inflammation and disease resistance of abalone Haliotis discus hannai

Clostridium autoethanogenum protein (CAP) is an eco-friendly protein source and has great application potential in aquafeeds. The present study aimed to investigate the effects of dietary CAP inclusion on the anti-oxidation, immunity, inflammation, disease resistance and gut microbiota of abalone Haliotis discus hannai after a 110-day feeding trial. Three isonitrogenous and isolipidic diets were formulated by adding 0 % (control), 4.10 % (CAP4.10) and 16.25 % (CAP16.25) of CAP, respectively. A total of 540 abalones with an initial mean body weight of 22.05 ± 0.19 g were randomly distributed in three groups with three replicates per group and 60 abalones per replicate. Results showed that the activities of superoxide dismutase and glutathione peroxidase in the cell-free hemolymph (CFH) were significantly decreased and the content of malondialdehyde in CFH was significantly increased in the CAP16.25 group. The diet with 4.1 % of CAP significantly increased the activities of lysozyme and acid phosphatase in CFH. The expressions of pro-inflammatory genes such as tumor necrosis factor-α (tnf-α), nuclear factor-κb (nf-κb) and toll-like receptor 4 (tlr4) in digestive gland were downregulated, and the expressions of anti-inflammatory genes such as β-defensin and mytimacin 6 in digestive gland were upregulated in the CAP4.10 group. Dietary CAP inclusion significantly decreased the cumulative mortality of abalone after the challenge test with Vibrio parahaemolyticus for 7 days. Dietary CAP inclusion changed the composition of gut microbiota of abalone. Besides, the balance of the ecological interaction network of bacterial genera in the intestine of abalone was enhanced by dietary CAP. The association analysis showed that two bacterial genera Ruegeria and Bacteroides were closely correlated with the inflammatory genes. In conclusion, the 4.10 % of dietary CAP enhanced the immunity and disease resistance as well as inhibited the inflammation of abalone. The 16.25 % of dietary CAP decreased the anti-oxidative capacity of abalone. The structure of the gut microbiota of abalone changed with dietary CAP levels.

Palliative effect of abalone insulin-related peptide 2 on emersion stress-induced hyperglycemia in the Pacific abalone (Haliotis discus hannai)

The Pacific abalone (Haliotis discus hannai), an economically important marine mollusk in East Asia, is often exposed to extreme environments, such as high water temperatures during summer and air exposure during handling or transportation. These stressful conditions lead to hyperglycemia and the generation of reactive oxygen species (ROS), resulting in oxidative stress and subsequent cell damage in the organism. To understand the role of insulin-related peptides in the regulation of elevated hemolymph glucose, hyperglycemia was confirmed in abalone under high water temperature and air (emersion) exposure (p < 0.05). To prove the existence of a glucose regulatory system in abalone, changes in hemolymph glucose were measured after bovine insulin injection, and hemolymph glucose was lowered by bovine insulin injection in hyperglycemia caused by both stressors (p < 0.05), supporting the existence of a glucose regulatory system functionally similar to that of vertebrates. Among the three abalone insulin-related peptides (AIPs), which were previously reported to be upregulated along with hemolymph glucose levels upon feeding, the relative mRNA expression of AIP2 in the cerebral ganglion showed a close correlation with high water temperature and emersion stress, which peaked 12 h after treatment (p < 0.05). Based on these results, we focused on the function of AIP2 in glucose homeostasis and characterized the localization of AIP2 mRNA-expressing cells in the cerebral ganglion using in situ hybridization. Positive signals from RNA probes were detected in neural cortex cells in the cerebral ganglion. Furthermore, suppressing AIP2 mRNA expression using RNA interference with double-stranded RNA (dsRNA) revealed that dsRNA-AIP2 injected abalone maintained high hemolymph glucose levels after 24 h of recovery following 1 h of emersion stress (p < 0.01). In contrast, abalones that received dsRNA-GFP and molluscan saline showed relatively rapid clearance of hemolymph glucose. This study demonstrates that AIP2 gene expression affects the recovery rate from emersion stress-induced hyperglycemia by regulating hemolymph glucose.

Structural and functional characterization of Hdh-HSBP1 and its involvement in heat stress and early development in Pacific abalone, Haliotis discus hannai

Heat shock factor binding protein 1 (HSBP1) is known to regulate the activity of heat shock factor 1 (HSF1) and the early development of organisms. To understand the involvement of HSBP1 in the heat shock response and embryonic and larval development of Pacific abalone (Haliotis discus hannai), the Hdh-HSBP1 gene was sequenced from the digestive gland (DG) tissue. The full-length sequence of Hdh-HSBP1 encompassed 738 nucleotides, encoding an 8.42 kDa protein consisting of 75 deduced amino acids. The protein contains an HSBP1 domain and a coiled-coil domain, which are conserved features in the HSBP1 protein family. Protein-protein molecular docking revealed that the coiled-coil region of Hdh-HSBP1 binds to the coiled-coil region of Hdh-HSF1. Tissue expression analysis demonstrated that the highest Hdh-HSBP1 expression occurred in the DG, whereas seasonal expression analysis revealed that this gene was most highly expressed in summer. In heat-stressed abalone, the highest expression of Hdh-HSBP1 occurred at 30 °C. Moreover, time-series analysis revealed that the expression of this gene began to increase significantly at 6 h post-heat stress, with higher expression observed at 12 h and 24 h post-heat stress. Furthermore, Hdh-HSBP1 mRNA expression showed a link to ROS production. Additionally, the expression of Hdh-HSBP1 showed significantly higher expression in the early stages of embryonic development in Pacific abalone. These results suggest that Hdh-HSBP1 plays a crucial role in the stress physiology of Pacific abalone by interacting with Hdh-HSF1, as well as its embryonic development.

Effectiveness of the garlic (Allium sativum) juice on removing the Pacific abalone postlarvae (Haliotis discus hannai) and subsequent growth evaluation

The Pacific abalone (Haliotis discus hannai) is widely cultivated in East Asian countries. Aquaculture of this species has become a thriving industry in China, with adequate seed production being a key contributing factor. However, maintaining sufficient diatom abundance during the nursery phase remains challenging. In this study, we assessed a transfer nursery culture method that involves prolonged cultivation with a sufficient diatom supply. This method involved treating postlarvae on diatom-depleted plates with a garlic juice (GJ) solution to facilitate detachment, followed by transfer to newly diatom-precoated plates for further growth in the nursery. We investigated the effects of garlic juice concentration (GJC) and animal size on detachment and mortality rates using a two-factor design. Our negative binomial regression analysis uncovered significant impacts of both factors and their interactions on the target traits. We noticed that the optimal GJC for smaller groups of abalone postlarvae (2.2 ± 1.0 mm) was 0.7 %, while for larger groups (3.8 ± 2.2 mm), it increased to 1.0 %. Notably, suitable GJC concentrations during post-treatment recovery resulted in no apparent mortality in Pacific abalone postlarvae. Our study demonstrated the effectiveness of the transfer nursery method, showing an improvement of over ∼1.1 mm in shell length and a 43.1 % increase in survival during a 30-day experiment. Our study presents a significant advancement, as it likely introduces the first reported method of large-scale postlarval removal of H. discus hannai for prolonged nursery periods with sufficient diatom provision. The integration of GJ treatment and the transfer nursery method exhibited promising results, highlighting the benefits of its application for enhancing husbandry procedures, nursery management, and the overall industrial productivity of Pacific abalone.

Dietary hydroxyproline supplementation improves the muscle texture and extends the shelf life of abalone Haliotis discus hannai

In the present study, an 84-day growth trial was conducted in abalone Haliotis discus hannai to evaluate the influences of dietary hydroxyproline (Hyp) levels (0.05, 0.53 and 1.24%) on the growth, collagen metabolism, muscle texture and its changes during storage at 4 °C. The results showed that the weight gain rate of abalones in the groups with 0.53% and 1.24% of dietary Hyp were significantly higher than that in the group with 0.05% dietary Hyp. The 0.53% and 1.24% of dietary Hyp significantly increased lysyl hydroxylase and prolyl 4-hydroxylase activity and pyridinium crosslink content, and up-regulated collagen biosynthesis-related gene expression in abalone muscle. Meanwhile, dietary Hyp supplementation significantly inhibited the mRNA expression of mmp-1 and significantly up-regulated the mRNA expression of timp. During the same storage time, the springiness, hardness and water-holding capacity of muscle in the dietary Hyp supplementation treatments were significantly higher. The muscle hardness in the dietary Hyp supplementation groups on day 6 was higher than that in the control group on day 3. Meanwhile, dietary Hyp significantly decreased the content of carbonyl and malonaldehyde, as well as the activity of cathepsin B and cathepsin L in the muscle of abalone during the storage. In conclusion, dietary Hyp supplementation increased collagen content, improved texture and extended the shelf life of abalone.

Total Iodine Quantification and In Vitro Bioavailability Study in Abalone (Haliotis discus hannai) Using Inductively Coupled Plasma Mass Spectrometry.

The aim of this study is to determine the total iodine content in Korean abalone (Haliotis discus hannai) and to investigate the bioavailability of iodine using an in vitro method. This research paper focuses on total iodine quantification in abalone (Haliotis discus hannai) and its components (viscera and muscle) using inductively coupled plasma mass spectrometry (ICP-MS). Additionally, an in vitro bioavailability study explored iodine absorption potential. Abalone pretreatment involved both the European standard method (ES) and microwave-assisted extraction method (MAE). The limits of detection (LOD) were 0.11 ng/g for both ES and MAE, with a limit of quantification (LOQ) of 5.4 ng/g for MAE. Accuracy, assessed using a reference material (fish muscle, ERM-BB422), showed values of 1.5 ± 0.010 mg/kg for ES and 1.6 ± 0.066 mg/kg for MAE, within an acceptable range of 1.4 ± 0.42 mg/kg. Precision, evaluated using the Horwitz ratio (HorRat) with a reference material, was determined to be 0.45 for ES and 0.27 for MAE. Therefore, total iodine contents were estimated as 74 ± 2.2 µg/g for abalone viscera and 17 ± 0.77 µg/g for abalone muscle with ES, and 76 ± 1.0 µg/g for abalone viscera and 17 ± 0.51 µg/g for abalone muscle with MAE. Recovery tests demonstrated an acceptable range of 90-110%. In the in vitro bioavailability assessment, digestion efficiency yielded ranges of 42-50.2% for viscera and 67-115% for muscle. Absorption efficiency variations were determined as 37-43% for viscera and 48-75% for muscle.

Ascorbic acid biosynthesis in Pacific abalone Haliotis discus hannai Ino and L-gulonolactone oxidase gene loss as an independent event

Up to now, it has been believed that invertebrates are unable to synthesize ascorbic acid (AA) in vivo. However, in the present study, the full-length CDs (Coding sequence) of L-gulonolactone oxidase (GLO) from Pacific abalone (Haliotis discus hannai Ino) were obtained through molecular cloning. The Pacific abalone GLO contained a FAD-binding domain in the N-termination, and ALO domain and conserved HWAK motif in the C-termination. The GLO gene possesses 12 exons and 11 introns. The Pacific abalone GLO was expressed in various tissues, including the kidney, digestive gland, gill, intestine, muscle and mantle. The GLO activity assay revealed that GLO activity was only detected in the kidney of Pacific abalone. After a 100-day feeding trial, dietary AA levels did not significantly affect the survival, weight gain, daily increment in shell length, and feed conversion ratio of Pacific abalone. The expression of GLO in the kidney was downregulated by dietary AA. These results implied that the ability to synthesize AA in abalone had not been lost. From the evolutionary perspective, the loss of GLO occurred independently as an independent event by matching with the genomes of various species. The positive selection analysis revealed that the GLO gene underwent purifying selective pressure during its evolution. In conclusion, the present study provided direct evidence to prove that the GLO activity and the ability to synthesize AA exist in abalone. The AA synthesis ability in vertebrates might have originated from invertebrates dating back 930.31 million years.

Characterization of myoinhibitory peptide signaling system and its implication in larval metamorphosis and spawning behavior in Pacific abalone

Myoinhibitory peptides (MIPs) affect various physiological functions, including juvenile hormone signaling, muscle contraction, larval development, and reproduction in invertebrates. Although MIPs are ligands for MIP and/or sex peptide receptors (MIP/SPRs) in diverse arthropods and model organisms belonging to Lophotrochozoa, the MIP signaling system has not yet been fully investigated in mollusks. In this study, we identified the MIP signaling system in the Pacific abalone Haliotis discus hannai (Hdh). Similar to the invertebrate MIPs, a total of eight paracopies of MIPs (named Hdh-MIP1 to Hdh-MIP8), harboring a WX5-7Wamide motif, except for Hdh-MIP2, were found in the Hdh-MIP precursor. Furthermore, we characterized a functional Hdh-MIPR, which responded to the Hdh-MIPs, except for Hdh-MIP2, possibly linked with the PKC/Ca2+ and PKA/cAMP signaling pathways. Hdh-MIPs delayed larval metamorphosis but increased the spawning behavior. These results suggest that the Hdh-MIP signaling system provides insights into the unique function of MIP in invertebrates.

Comparison of Growth, Gut Microbiota Composition, and Taste Sensory Profile of Abalone (Haliotis discus hannai) Fed Diets Formulated With Heat-Treated Or Fermented Seaweed (Sargassum horneri) Meal

Comparison of Growth, Gut Microbiota Composition, and Taste Sensory Profile of Abalone (Haliotis discus hannai) Fed Diets Formulated With Heat-Treated Or Fermented Seaweed (Sargassum horneri) Meal

Effects of reproductive condition on haemocyte parameters in the Pacific abalone Haliotis discus hannai during the spawning period

ABSTRACT The assessment of morphology and immunology of specimens at different maturity stages is essential for the determination of spawning strategy and reducing the risk of mortality. In this paper, female and male Pacific abalones (Haliotis discus hannai) at different statuses (pre-spawning, spawning and post-spawning) were collected to estimate the effects of spawning on the immune response. The haemocyte parameters, including total haemocyte counts (THC), haemocyte mortality, and reactive oxygen species production (ROS) were analysed using flow cytometry. THC decreased after spawning, which was lowest 3 days post-spawning. Haemocyte mortality was higher during spawning, and was highest 3 days post-spawning. Abalone also reduced their metabolic activity after spawning, reflected by an increase in ROS in both females and males. Moreover, our results show that female abalone reach the spawning period with a higher degree of activities of haemocytes and immune responses than male. The results confirmed that variability in abalone haemocytes during the spawning period might related to individual metabolic conditions.