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This study evaluated the effects of Gracilariopsis lemaneiformis hydrocolloids on Nile tilapia (Oreochromis niloticus) using an advanced multiomics approach (transcriptome and proteome) linked with genomic isoform structure to elucidate the biofunctions of G. lemaneiformis hydrocolloids. The results showed that G. lemaneiformis hydrocolloids did not affect growth, as indicated by the nonsignificant differences in growth and blood biochemical indicators.Regarding the response, both intestine and liver tissues were assessed. These findings indicate that 20 % G. lemaneiformis hydrocolloids enhanced cytokine expression, which may contribute to a biological function in the intestine and liver of O. niloticus.Genome and proteome profiles indicated that G. lemaneiformis hydrocolloids upregulated the intestine and liver peroxisome proliferator-activated receptor (PPAR) signaling pathway, nucleocytoplasmic transport, steroid biosynthesis, and histidine metabolism. In contrast, co-factor biosynthesis, nucleocytoplasmic transport, tryptophan metabolism, arginine and proline metabolism, arginine biosynthesis, and ribosome activity were downregulated. These findings indicate that G. lemaneiformis hydrocolloids significantly affect liver lipid and carbohydrate metabolism. Proteomics analysis revealed that G. lemaneiformis hydrocolloids upregulated the PPAR signaling pathway, playing a crucial role in lipid metabolism.In summary, 20 % G. lemaneiformis hydrocolloids are primarily involved in modulating the intestine and liver PPAR signaling pathway to regulate lipid metabolism.

ABSTRACT Understanding the functions of mitochondrial genomes is crucial for studies related to the evolution of genomes, phylogenomics, and species identification. For the first time, complete mitogenome of Potamalpheops sp. a non‐snapping shrimp has been successfully sequenced and characterized from Taiwan that belongs to the Decapoda order, Crustacea class, and Caridea infraorder. This study involved analysis of nucleotide composition, codon usage, gene ordering, evolutionary selection pressure, and comparative mitogenomics. The mitogenome of Potamalpheops sp. is 16,605 base pairs in length and consists of standard set of 37 genes found in metazoans. The gene rearrangements in the mitochondrial genome of this species shows extensive rearrangements comparing to the typical pattern found in pancrustaceans mitogenomes. Therefore, it could be concluded that gene rearrangements most likely happen only in the caridea infraorder. The current investigation discovered transposition of the tRNA and rRNA genes along with reversal in strands in the tRNAs. No other Alpheidae mitochondrial genome that has been investigated thus far has revealed this pattern. All 13 protein coding genes in the mitochondrial genomes of superfamily Alphoidea exhibited Ka/Ks values lower than 1, according to the ratios of nonsynonymous and synonymous substitutions rates. This suggests that a strong purifying selection had taken place. The maximum likelihood tree consisting of 46 mitogenomes of infraorder Caridea along with outgroups, revealed the existence of Potamalpheops sp. in the family Alpheidae and it formed a monophyletic group along with Palaemonoidea and Alpheoidea superfamily.

The largescale blackfish, Girella punctata (Gray, 1835), inhabits rocky substrates in the coastal waters of Japan and northeastern Taiwan. Despite its commercial significance, there is limited knowledge regarding its reproductive biology due to the sparse occurrence of broodstocks in its natural environment. This study investigated the reproductive performance and embryonic development of G. punctata, assesses its feeding preferences through controlled feeding trials, and evaluates the impact of various feed types on its growth performance. In total, 124 wild-caught G. punctata were reared for 3–4 years, and their reproductive performance was assessed in 2015 and 2016. Their egg production increased with a sudden water temperature change from 17 ℃ in 2015–22 ℃ and 2016. The total and daily average egg production weights were 3589.22 and 40.79 g, respectively, whereas the total and daily average fertilized egg production weights were 1630.72 and 18.59 g, respectively; moreover, the hatching rate was 42.61 %. During embryonic development, the eggs hatched 45 h post fertilization at 19 ℃. Newly hatched larvae were 2.26 ± 0.04 mm in length with a yolk sac of 1.09 ± 0.02 mm in length. The yolk sac was almost completely absorbed 3 days post hatch (dph), when the larvae grew to a length of 3.42 ± 0.06 mm. We found that fish meal with 40 % protein was the optimal G. punctata fish diet. The weight gain in juveniles fed Cladophora fascicularis ranged from 47 % to 52 %, which was significantly higher than that in those fed Hypnea charoides (p < 0.05). The present study provided a practical guideline on the conditions required for successful fertilization and early development of G. punctata embryos and other larval stages. This information should help ensure greater survival and a higher proportion of healthy G. punctata larvae for mariculture applications.

The small abalone (Haliotis diversicolor) is commercially valuable for fisheries and aquaculture. Once ranked first globally, Taiwan's abalone industry is now on the brink of collapse due to disease issues. Despite improvements in survival rates through crossbreeding between Japan and Taiwan populations, the subsequent lack of further breeding selection and genetic management has led to disorder after over a decade of introduction. With frequent commercial exchanges, stock enhancement, private introductions, and hybridization, genetic management of H. diversicolor poses both challenges and necessity. This study utilizes ISSRseq technology to aid in the genetic management of H. diversicolor. Various analyses show significant differences among the Japan wild population, Taiwan wild populations, the Taiwan-Indonesia hybrid strain, and other cultured populations. Selective strains bred by research institutions and private hatcheries exhibit uniqueness distinct from commercial H. diversicolor. The presence of multiple populations exhibiting admixture may be attributed to improper stock enhancement and management practices. Our work elucidates the current status of genetic resources of Taiwan H. diversicolor, laying the foundation for future breeding efforts.

This study investigated the effect of substances on nonspecific immune responses of head kidney leukocytes, the antimicrobial activity against Vibrio sp., as well as the time-kill of Vibrio sp. by combining the substances with lactic acid bacteria (LAB) and Pediococcus sp. The substances are B. pilosa hot water extract, B. pilosa powder extract, B. pilosa methanol extract, B. pilosa ethanol extract, B. alba hot water extract, B. alba powder extract, B. alba methanol extract, B. alba ethanol extract, and bioactive compounds, namely cytopiloyne, flavonoid, phenol, ethyl caffeate, luteolin, chlorogenic acid, butein, and linoleic acid. The results showed that some of them were nontoxic to the head kidney leukocytes, which can increase the phagocytic rate, phagocytic index, and respiratory burst. These substances were able to inhibit the growth of Vibrio sp.; they can even completely kill the pathogenic bacteria. The largest of the inhibition zone formed from the EC group at a concentration range of 5–50 µg/mL against V. parahaemolyticus, V. alginolyticus, and V. harveyi with a value of 19.7 ± 0.56, 19.3 ± 1.53, and 20.6 ± 1.53 mm. Furthermore, the time-kill studies showed that the LAB and P. acidilactici can completely kill the Vibrio sp. at 6 h incubation time, mainly in the group of combination with EC.

Beehorn snail (Clithon diadema), known for its vibrant colors, is threatened with extinction due to excessive field collection. Therefore, this study investigated the effects of different diets and salinity (0‰–35‰) on the gonadal development, capsule productivity, embryonic development, and larval survival and growth of C. diadema. Adult snails (shell length: 9.09 ± 0.87 mm) were fed diatoms, including pennate marine diatom (Nitzschia grossestriata), boat-shaped diatom algae (Navicula sp.) and Cylindrotheca sp., along with giant tiger prawn (Penaeus monodon) commercial feed, sea lettuce (Ulva lactuca), or Cladophora fascicularis, while snail larvae (shell length: 154.85 ± 6.03 μm) were fed Isochrysis galbana or Chaetoceros muelleri. The diatom group outperformed commercial feed and U. lactuca groups. The highest capsule productivity, yielding 79–82 capsules in 4 weeks, occurred at 15‰-20‰ salinity for diatom-fed adults. Embryos thrived at 0‰ but didn't hatch; hatching required salinities above 0‰, highlighting saline necessity. In addition, acute salinity changes (0‰ to 30‰) can deform embryo. Embryonic development at 15‰–20‰ salinity lasted 13.56–14.33 days with 100% larval hatching. However, larvae hatched at >20‰ salinity exhibit improved the percentage of survival without feeding (longer median lethal times). I. galbana-fed larvae survived 68 days with a 75% rate, outlasting C. muelleri-fed larvae which perished within 28 days. I. galbana-fed larvae, raised for 68 days at 25‰ salinity, showed 100% survival, 44.44% metamorphosis, and a 3.1 times growth increase. Metamorphosis rates were highest at 25‰ salinity with I. galbana, exceeding the 20‰ group by 3.2 times. Mucus of C. diadema boosts larval metamorphosis to 50.00%, 9 times higher than without inducers. Based on previous results, we developed an artificial propagation and breeding method for C. diadema. Artificially propagating C. diadema in large quantities could reduce the need for wild harvesting, thereby mitigating negative ecological impacts from excessive collection.

This study investigated the impact of Galla chinensis (GC) powder as a dietary supplement on the immunity, growth, and pathogen resistance of white shrimp, Penaeus vannamei. An in vitro analysis demonstrated that GC inhibited Vibrio with no discernible impact on Lactobacillus. Following this, GC was tested for toxicity on white shrimp hemocytes. GC showed harmlessness to white shrimp hemocytes since cell viability was over 80% at all dosages. The immune response of hemocytes in white shrimp to diets containing varying concentrations of GC was evaluated in an in vivo experiment. Five feeding regimens were trialed for 28 days, which were designated as GC0.5, GC1, GC2.5, and GC5 (0, 0.5, 1, 2.5, and 5 g/kg, respectively). The enhanced total hemocytes, phenoloxidase, phagocytic activity, and production of superoxide anion in white shrimp on day 7 of the feeding trial by administering a supplementary GC diet significantly stimulated the nonspecific immunity of white shrimp. Additionally, dietary GC substantially increased the expression of related genes in white shrimp hemocytes in vivo, including Toll, Imd, antimicrobial peptides, antioxidant defenses, clotting protein, and heat shock protein. The challenge experiment was conducted to assess the disease resistance of white shrimp fed with GC. The shrimps were fed dietary GC for 7 days while being challenged with Vibrio parahaemolyticus. The shrimp exhibited a substantial variation in survival rate during the bacteria challenge experiment due to the activation of related genes and the enhancement of non-specific immunity. The mortality of white shrimp was substantially reduced by the supplementary GC at a concentration of 1 g/kg during the bacteria challenge. A final feeding trial was conducted to assess the effects of dietary GC on the growth performance of white shrimp. The continuous administration of dietary GC at a dose of 1 g/kg to shrimp for 56 days resulted in the highest enhancement in shrimp growth among all treatments. This was evidenced by significantly increased final body weight, weight gain, specific growth rate, and decreased feed conversion ratios. Dietary GC at a dosage of 1 g/kg has been suggested for use to enhance the growth performance of white shrimp and improve immunological responses, particularly nonspecific immunity and related gene expression, as well as resistance to Vibrio parahaemolyticus.

Abstract Growth selection in fish is essential for aquaculture; however, long generation times and artificial breeding limitations present challenges for breeding endeavors. The tomato grouper (Cephalopholis sonnerati) is an emerging aquaculture species with a generation cycle of at least 5 to 10 years. Mixed culture of broodstocks and natural mating are commonly employed in hatcheries. To expedite the selection process for tomato grouper, we utilized ISSRseq technology for assisted future growth selection. After 6 months of rearing, we selected 48 individuals, including 24 fast‐growing and 24 slow‐growing individuals (representing the top and bottom 10% of samples). These samples produced 932 SNPs. Although specific broodstock pairing information is unavailable, our results suggest that these fry likely originated from multiple broodstock exhibiting considerable diversity. Further analysis of genetic relationships among individuals revealed three main clades. One clade contained a higher proportion of faster‐growing individuals (80%), while another featured a lower proportion of rapidly growing individuals (28.6%). Through the relatedness network and further exclusion of individuals, the faster‐growth clade comprised 92.3% of fast‐growing individuals. The faster‐growth clade is appropriate for further development into a high‐growth strain. Our findings demonstrate that molecular markers can advance population selective breeding to family selection in aquaculture breeding research.

The health of the host is significantly influenced by the gut microbiota. Penaeus vannamei (white shrimp) is one of the most profitable aquaculture species globally. Synbiotics are typically used as a beneficial diet supplement for raising aquaculture species’ growth capacities and enhancing immunity against pathogenicity. However, the effects of synbiotics on the white shrimp intestinal microbiota remain poorly understood. In the present study, we targeted the V3–V4 region of 16S rRNA genes to analyze the effects of synbiotics on white shrimp gut microbiota. Dietary synbiotics, having Lactobacillus acidophilus, and Moringa oleifera leaf extract were added to the white shrimps’ feed in various proportions in the present study. In total, 490 operational taxonomic units yielding 23 phyla, 41 classes, 94 orders, 151 families, and 250 genera of microorganisms were obtained. The diet containing L. acidophilus at 1 × 107 CFU/g and M. oleifera at 2.5 g/kg led to an increase in the relative abundance of beneficial microorganisms through a significant decrease in the α diversity. Moreover, it upregulated several physiological pathways such as carbohydrate metabolism, signal transduction, lipid metabolism, nucleotide metabolism, amino acid metabolism, and environmental adaptation, which led to the upregulation of the AMPK, MAPK, P13K-Akt, lysosome, peroxisome, and ferroptosis signaling pathways; this enhanced growth and immunity in white shrimp. Whether a single species or a combination of different microorganisms improves growth and immunity remains unclear till now. Nevertheless, our results will facilitate further in-depth investigation into beneficial microbial communities for upliftment of white shrimp aquaculture.

Extracellular vesicles (EVs) are functional substances secreted by microbes and host cells, and it has been discovered that they participate in the interactions between different microorganisms. Our recent findings indicate that Limosilactobacillus reuteri-derived EVs have the potential to improve the intestinal microbiota of Oplegnathus fasciatus fish and inhibit pathogenic bacteria. Previous research has reported that the host intestinal cells play a regulatory role in the intestinal microbiota. This suggested that to investigate the mechanisms through which L. reuteri-derived EVs regulate the intestinal microbiota, a system that excludes interference from host intestinal cells should be established. In this study, an in vitro cultured intestinal bacteria system, without host factors, was used to simulate the intestinal microbiota of O. fasciatus fish. After adding L. reuteri-derived EVs to the system, the changes in the microbiota were analyzed. The results showed that L. reuteri-derived EVs effectively reduced the abundance of Vibrio spp. In the results of the in vitro experiments, it was also observed that L. reuteri-derived EVs have the ability to inhibit Vibrio alginolyticus. We further sequenced the small RNA contained in L. reuteri-derived EVs and found that these small RNAs can interfere with genes (LysR, pirin, MIpA/OmpV, CatB, and aspartate-semialdehyde dehydrogenase) related to the growth of V. alginolyticus. Taken together, the results indicate that in the absence of host involvement, the small RNAs present in L. reuteri-derived EVs have the function of inhibiting pathogenic bacteria and exhibit the potential to regulate the intestinal microbiota.

Discoloration is a critical limitation in the storage of fish meat. In this study, the antioxidant and antimicrobial activities of phytochemicals extracted from mango leaf, guava leaf, and green tea residue and a mixture of these phytochemicals (1:1:1, w/w/w) were initially evaluated in vitro. Compared with the individual byproducts, the mixture exhibited higher antioxidant and antimicrobial activities and was selected for use as a feed or processing additive. The half-maximal inhibitory concentration of the phytochemicals mixture for 2,2-diphenyl-1-picrylhydrazyl scavenging activity was calculated as 10.46 μg/mL. In addition, the minimum inhibitory concentration and minimum bactericidal concentration of the mixture against Aeromonas hydrophila, Edwardsiella tarda, Escherichia coli, Lactococcus garvieae, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus iniae ranged from 0.20 to 12.50 mg/mL. Nile tilapia were fed with diets containing different concentrations of the phytochemicals mixture, namely 0.0% (control), 0.5%, 1.0%, and 2.0%, for a duration of 8 weeks before filleting. The results indicated that the growth performance of the fish was not affected by the phytochemicals mixture. In the immersion treatment, half of the fillets from the control group were immersed in a solution containing a 1 mg/mL phytochemicals mixture and air-dried. During the storage period, chromatic analysis revealed a stable increase in the CIE b* value, with both dietary and immersion treatments significantly inhibiting discoloration (p

Abstract Astaxanthin (AST), a super antioxidant with coloring and medical properties, renders it a beneficial feed additive for shrimp. This study conducted a white shrimp feeding trial of 3S, 3’S isoform AST, which was derived from metabolic-engineered Kluyveromyces marxianus fermented broth (TB) and its extract (TE) compared to sources from two chemically synthetic ASTs (Carophyll Pink [CP] and Lucantin Pink [LP]), which contain 3S, 3’S, 3R, 3’S (3S, 3’R) and 3R, 3’R isoforms ratio of 1:2:1. The effects on red coloration, immune parameters and resistance to Vibrio infection were evaluated. Four AST sources were incorporated into the diets at concentrations of 0 (control), 100 mg kg−1 (TB100, TE100, CP100, and LP100), and 200 mg kg−1 (TB200, TE200, CP200, and LP200). Results revealed that in week 4, shrimps that received AST-supplemented feeds, especially TB100, TB200, and TE200, significantly increased redness (a*) values. Immune responses including phagocytosis activity, superoxide-anion production, phenoloxidase activity, and immune-related genes were examined on days 0, 1, 2, 4, 7, 14, 21, and 28. Generally, shrimps that received AST-supplemented feeds exhibited higher immune responses on days 7 and 14 than the control feed. Gene expression levels of superoxide dismutase and glutathione peroxidase were significantly upregulated on days 7 and 14 in shrimps that received AST-supplemented feeds, while genes of penaeidins, antilipopolysaccharide factor, and lysozyme were upregulated on days 4, 7, and 14, especially received TB200 and TE200. Furthermore, shrimps that received TB100, TE100, CP100, and LP100 7 days were then challenged with Vibrio parahaemolyticus and the result demonstrated higher survival rates especially TB100 at 168 h than the control feed. In conclusion, incorporating AST into the diets enhanced shrimp red coloration, immune parameters, and resistance against V. parahaemolyticus infection. The K. marxianus-derived AST exhibited higher performance than did chemical AST to be a potential feed additive in shrimp aquaculture.

The habitats of giant clams are undergoing environmental changes, and giant clam populations are declining. The present study was conducted to facilitate clam conservation. We conducted three 18-week trials to investigate the effects of nutrient, temperature, and salinity on the growth performance and survival rates (SRs) of juvenile Tridacna noae, adult Tridacna crocea, and subadult Tridacna derasa, respectively. Regarding nutrient sources, no significant differences were observed in shell length gain, specific growth rate, or SR between clams fed with Chaetoceros muelleri or commercial feed (hw nanotip) and those in a control group (juvenile phototrophs). Regarding temperature, clams cultivated at 27 °C exhibited significantly better growth performance and SR than did those cultivated at 19 °C or 31 °C (p < 0.05). By week 6, all clams in the 19 °C and 31 °C groups had died, indicating that suboptimal growth temperatures have severe adverse effects. Regarding salinity, clams cultivated at 34‰ exhibited significantly higher length gains and specific growth rates than did those cultivated at 20‰ or 25‰ (p < 0.05). SR was not significantly affected by salinity. Understanding how environmental factors affect giant clam populations may help researchers devise effective clam conservation strategies.

In recent years, probiotics have received considerable attention for improving the health of aquaculture organisms, such as fish and shrimp, by stimulating immune activity and increasing growth rates. Oplegnathus fasciatus is a common and economically important cultured fish species in Asia. In this study, we aimed to investigate the potential of lactic acid bacteria (LAB; Limosilactobacillus reuteri)-fermented feed to promote growth and enhance immune function in O. fasciatus. The feed contained the highest proportion of LAB after L. reuteri fermentation for 3 days in anaerobic conditions. Oplegnathus fasciatus was fed LAB-fermented feed for 30 days. The administration of LAB-fermented feed (live bacteria > 109 CFU/g) significantly increased the growth rate (weight gain = 174.8%; FCR = 4.23) and intestinal probiotic levels of O. fasciatus. After LAB-fermented feeding, the immunity index was evaluated by superoxide anion production, the phagocytic activity of leukocytes, and bactericidal and lysozyme activities in the serum of O. fasciatus. We found that LAB-fermented feed treatment potentially elevated the proportions of intestinal Bifidobacterium, Blautia, and Dorea species and reduced pathogenic bacterial growth (Acinetobacter, Escherichia_Shigella, and Megasphaera) in O. fasciatus. This study demonstrated that LAB-fermented feed containing extracellular vesicles improves growth performance and the inhibition of pathogenic Acinetobacter baumannii.

Fish processing byproducts are conventionally used in live feed production for larviculture because of their low cost and high productivity. However, waste-generated bacteria may cause pathogen proliferation, and antibiotics are widely used to reduce the bacterial loads of live feeds. In this study, tilapia processing byproducts were hydrolyzed into fish protein hydrolysate (FPH). Subsequently, the FPH was used to culture Rhodopseudomonas palustris, a photosynthetic bacterium (PSB), as a nutritional source for live feed. The results of the in vitro pathogen inhibition assay indicated that R. palustris significantly inhibited the growth of Aeromonas hydrophila, Edwardsiella tarda, Lactococcus garviae, and Streptococcus iniae in the FPH supernatant (p

The water environment plays an important role in animal physiology. In this study, we sought to evaluate the effect of the acid environment on the Oreochromis niloticus (Nile tilapia) internal microenvironment immune response compare to the mouse macrophage model (J77A.1). The acid environment treated mouse macrophage J774A.1 model have shown that acidic treatment is able to polarize macrophages into M2-like macrophages via an increase in Ym1, Tgm2, Arg1, Fizz1, and IL-10 expression. Metabolic analysis of mouse macrophages (J774A.1) at pH 2 vs. pH 7 and pH 4 vs. pH 7 have been shown to promote the expression of intracellular acetylcholine, choline, prochlorperazine, L-leucine, and bisphenol A,2-amino-3-methylimidazo[4,5-f] quinolone metabolites in the M2-like macrophage. Immune gene expression of the O. niloticus spleen and liver treated at pH 2, 4, and 7 was shown to reduce TNF-α, IL-1 β, IL-8, and IL-12 expression compared to pH 7 treatment. Immune gene was induced in O. niloticus following culture at pH 5, 6, and 7 fresh water environment. Taken together, we found that the acid internal environment polarizes tissues into an M2 macrophage developmental microenvironment. However, if the external environment is acid, tissues are exposed to an M1 macrophage developmental microenvironment.

Malachite green (MG) residue in aquatic environments can adversely affect food chains. To solve this problem, many adsorbents have been developed to remove MG from aqueous solutions. These adsorbents exhibit benefits in different aspects such as low cost, nontoxicity, and environment friendliness. Zeolite, oyster shell powder (OSP), humic acid (HA), and Chloride dioxide (ClO2) are widely used agents for water purification, indicating their potential to remove MG from both freshwater and seawater. Liquid chromatography tandem mass spectrometry was performed as a valid analytical method for measuring MG and its metabolite leucomalachite green (LMG). The analysis examined linearity, specificity, accuracy, precision, and limit of quantification. We found that three agents—zeolite, OSP, and HA—have high affinity for MG and LMG in both freshwater and seawater because of their physicochemical properties; ClO2 exhibited even greater efficiency for MG and LMG degradation due to its oxidation capacity. The MG removal efficiency of zeolite, OSP and HA in freshwater was 100%, 87.52%, and 100%, respectively. In the MG-containing seawater, the removal efficiency of zeolite, OSP and HA was 95.24%, 6.57%, and 89.36%, respectively. The LMG removal efficiency of zeolite, OSP and HA in freshwater was 100%, 53.56%, and 100%, while in seawater, the LMG removal efficiency of zeolite, OSP and HA was 100%, 62.70%, and 84.65%, respectively. ClO2 treatments in both freshwater and seawater expressed 100% removal efficiency. The adsorption capacity of three water agents in MG-containing freshwater decreased in the order: zeolite = HA > OSP; in MG-containing seawater: zeolite > HA > OSP. Whereas the adsorption capacity of three water agents in LMG-containing freshwater decreased in the order: zeolite = HA > OSP; in LMG-containing seawater: zeolite > HA > OSP. In this study, four water treatment agents are reported to remove MG and LMG from freshwater and seawater, indicating their potential for reducing the amount of hazardous MG and LMG residue in aquatic environments.

An 8-week feeding trial was conducted to investigate the effects of replacing fishmeal with defatted black soldier fly larvae meal (DBSFLM) in the diets of Japanese eel on their growth performance, fillet texture, serum biochemical parameters, and intestinal histomorphology. Six isoproteic (520 g kg−1), isolipidic (80 g kg−1), and isoenergetic (15 MJ kg−1) diets were formulated with fishmeal replacement levels of 0% (R0), 15% (R15), 30% (R30), 45% (R45), 60% (R60), and 75% (R75). The growth performance, feed utilization efficiency, survival rate, serum liver function enzymes, antioxidant ability, and lysozyme activity of fish were not affected (P>0.05) by DBSFLM. However, the crude protein and cohesiveness of the fillet in groups R60 and R75 significantly decreased, and the fillet hardness significantly increased (P

Cobia (Rachycentron canadum) is a marine fish of high economic value that grows at a fast rate. However, intensive fish farming has led to disease outbreaks in cobia cultures, which is highly costly to the industry. The impact of infectious diseases on cobia production has led to the inappropriate and increased use of chemicals and antibiotics, which negatively affects the environment and human health and promotes the spread of drug-resistant pathogens. Hence, prophylactic measurements, such as the use of immunomodulators, are required to improve the health of cultured animals against pathogens. In this study, we examined the effects of Sarcodia suiae water extract (SSWE) in cobia in vitro and in vivo. We found that treatment with SSWE could significantly increase the expression of cytokines (e.g., IL-1β, IL-6, IL-10, IL-12, and TNF-α) and chemokines (e.g., IL-8) in primary cultured head kidney leukocytes. Intraperitoneal injection of SSWE (20 μg/g body weight) promoted higher expression of IL-6, IL-8, IL-10, IL-12, chemokines (e.g., CC1), and antibodies (e.g., IgT) in head kidney and spleen tissues of the fish compared with other dose levels. Additionally, we describe for the second time (only after India) of the isolation of Photobacterium damselae subsp. damselae (Phdd) from a deadly epizootic in cage-farmed cobia. An intraperitoneal inoculation of SSWE before Phdd challenge showed that SSWE treatment could delay the onset of mortality of cobia. Finally, fish that received SSWE intraperitoneally before infection with Phdd exhibited elevated expression of Th1-type cytokines, namely, IL-8, IL-12, TNF-α, and IFN-γ. At the same time, the expression of Th2-related factors (such as IL-10 in the head kidney, and IgM and IgT in the spleen) were lower for the fish that received SSWE instead of PBS before the Phdd challenge. The results indicate that SSWE treatment facilitates the induction of Th1-type cytokines in cobia to fight against Phdd infection and has the potential to be used as an immunostimulant and vaccine adjuvant for fish.

Fungi-derived β-glucan, a type of glucopolysaccharide, has been shown to possess immune-modulatory properties in clinical settings. Studies have indicated that β-glucan derived from Ganoderma lucidum (commonly known as Reishi) holds particular promise in this regard, both in laboratory and in vivo settings. To further investigate the efficacy and safety of Reishi β-glucan in human subjects, a randomized, double-blinded, placebo-controlled clinical trial was conducted among healthy adult volunteers aged 18 to 55. Participants were instructed to self-administer the interventions or placebos on a daily basis for 84 days, with bloodwork assessments conducted at the beginning and end of the study. The results of the trial showed that subjects in the intervention group, who received Reishi β-glucan, exhibited a significant enhancement in various immune cell populations, including CD3+, CD4+, CD8+ T-lymphocytes, as well as an improvement in the CD4/CD8 ratio and natural killer cell counts when compared to the placebo group. Additionally, a statistically significant difference was observed in serum immunoglobulin A levels and natural killer cell cytotoxicity between the intervention and placebo groups. Notably, the intervention was found to be safe and well tolerated, with no statistically significant changes observed in markers of kidney or liver function in either group. Overall, the study provides evidence for the ability of Reishi β-glucan to modulate immune responses in healthy adults, thereby potentially bolstering their defense against opportunistic infections.

Nostoc commune has abundant bioactive compounds, which are valuable in medical, cosmeceutical, and nutraceutical fields. Currently, the demand for native bioactive compounds in the cosmetic industry is greater than ever, especially with regard to algae extracts. In this study, we evaluated the biochemical compounds, safety, and effect on wound-healing and anti-allergic properties of Nostoc commune polysaccharide-rich extract (NCPE) in an in vitro study. The results showed that an extract with 221.69 ± 25.56 mg g−1 polysaccharide mainly consists of glucose (73.2%) with abundant bioactive compounds (total phenolic, total flavonoids, and sulfates) and some elements (K, Sr, Mn, Ba, and Ni). Cytotoxicity and heavy metal assays revealed that the extract was safe and available. Cell culture experiments showed that the polysaccharide effectively inhibits IL-6, and β-hexosaminidase, but promotes collagen I secretion. These findings suggest that the extract may be a useful resource in the development of wound-healing and anti-allergic cosmetics.

Bioactive compounds extracted from natural renewable sources have attracted an increased interest from both industry and academia. Recently, algae have been highlighted as promising sources of bioactive compounds, such as polyphenols, polysaccharides, fatty acids, proteins, and pigments, which can be used as functional ingredients in many industrial applications. Therefore, a simple green extraction and purification methodology capable of recovering biocompounds from algal biomass is of extreme importance in commercial production. In this study, we evaluated the application of three valuable algae (Colaconema formosanum, Sarcodia suae, and Nostoc commune) in combination with Pseudoalteromonas haloplanktis (type strain ATCC 14393) for the production of versatile compounds. The results illustrate that after 6 h of first-stage fermentation, the production of phycobiliproteins in C. formosanum was significantly increased by 156.2%, 188.9%, and 254.17% for PE, PC, and APC, respectively. This indicates that the production of phycobiliproteins from algae can be enhanced by P. haloplanktis. Furthermore, we discovered that after S. suae and N. commune were fermented with P. haloplanktis, mannose was produced. In this study, we describe a feasible biorefinery process for the production of phycobiliproteins and mannose by fermenting marine macroalgae with cyanobacteria. We believe it is worth establishing a scale-up technique by applying this fermentation method to the production of phycobiliproteins and mannose in the future.

Aquaculture plays a key role in food production globally and provides a valuable source of protein and nutrition, addressing a worldwide growing demand. Oplegnathus punctatus (spotted knifejaw) is an economically important fish species with a high market value and demand. Previous studies on O. punctatus focused mainly on gonadal development, chromosomal microstructure, selective breeding, characterization of immune genes, and viral diseases. There is no published scientific research regarding vibriosis in this fish species. In this study, two potential pathogenic bacteria, Vibrio harveyi and Enterococcus gallinarum, were isolated from moribund cultured O. punctatus. The sequence of the universal 16S rDNA gene was used to identify potential pathogenic bacteria isolated from the moribund O. punctatus, and morphological assessments and API20E tests of the bacterial isolates were conducted to verify the identity and biochemical characteristics of the isolates. Injection of E. gallinarum did not lead to mortality in O. punctatus during the 21 days of observation. In contrast, fish died overnight when challenged with V. harveyi at 1.25 × 105 CFU/g body weight, suggesting that the cause of death of the cultured O. punctatus was V. harveyi infection. Antimicrobial sensitivity analyses revealed that the V. harveyi strain NTOU is sensitive to flumequine, doxycycline, oxolinic acid, and amoxycillin. Importantly, we demonstrated for the first time that intraperitoneal administration of an inactivated V. harveyi whole-cell vaccine resulted in a high level of protection against V. harveyi infection in O. punctatus.

This study aimed to investigate the effects of dietary natural feed supplement on the growth performance, non-specific responses, and disease resistance in Lates calcarifer. Three commercial products (A, B, and C) containing a basal probiotic mixture were tested. Product A contained a basal mixture of Paenibacillus sp., Bacillus subtilis, Bacillus amyloliquefaciens, and Lactobacillus rhamnosus (107 cfu/g for each probiotic); product B contained additional Lactobacillus plantarum (1010 cfu/g); and product C contained additional soybean peptides (500 g/kg) and garlic powder (1 g/kg). Each product was supplemented into subject diets at dosages of 1 or 2 g/kg (designated as the A1, A2, B1, B2, C1, and C2 groups, respectively). Following an eight-week trial, growth parameters (specific growth rate and feed conversion ratio), non-specific immune responses (O2− production, phagocytic rate, and phagocytic index), and the results of a challenge test against Vibrio alginolyticus were evaluated. The results show that all probiotic supplement groups exhibited an improvement in growth performance compared to the control group (non-probiotic diet). In terms of non-specific immunity parameters, a significant improvement in O2− production was found in the C2 group, whereas significant improvements in phagocytic activity were found in all the B and C groups. The C2 group displayed optimal O2− production, phagocytic rate, and phagocytic index results. For the challenge test, the C groups showed higher Vibrio resistance than the other experimental groups and the control group. These results suggest that product C, given at dosages of 2 g/kg, may serve as a growth-promoting and immunostimulatory additive for the cultivation of Asian seabass.

The ocean resources have been rapidly depleted in the recent decade, and the complementary role of aquaculture to food security has become more critical than ever before. Water quality is one of the key factors in determining the success of aquaculture and real-time water quality monitoring is an important process for aquaculture. This paper proposes a low-cost and easy-to-build artificial intelligence (AI) buoy system that autonomously measures the related water quality data and instantly forwards them via wireless channels to the shore server. Furthermore, the data provide aquaculture staff with real-time water quality information and also assists server-side AI programs in implementing machine learning techniques to further provide short-term water quality predictions. In particular, we aim to provide a low-cost design by combining simple electronic devices and server-side AI programs for the proposed buoy system to measure water velocity. As a result, the cost for the practical implementation is approximately USD 2015 only to facilitate the proposed AI buoy system to measure the real-time data of dissolved oxygen, salinity, water temperature, and velocity. In addition, the AI buoy system also offers short-term estimations of water temperature and velocity, with mean square errors of 0.021 °C and 0.92 cm/s, respectively. Furthermore, we replaced the use of expensive current meters with a flow sensor tube of only USD 100 to measure water velocity.

Ultraviolet (UV-C) irradiation is the most important part of water filtration, which has no side effects on the environment and has been used in water purification systems in the aquaculture and transistor industries. In this research, the effect of UV-C on Chlorella sp. was investigated. Chlorella sp. was irradiated 0, 1, 2 or 3 times at a fixed flow rate of 6.5 L min−1 and the effects of UV-C LED on the apoptosis rate and death rate of Chlorella sp. were analyzed by flow cytometry after staining cells with the nucleic acid dye SYTOX Green and the membrane-associated protein stain Annexin V-PE Reagent. As a result of UV-C irradiation, the Chlorella sp. cells underwent phosphatidylserine (PS) ectropion and plasma membrane damage, which resulted in death. The effect of UV-C was proportional to the number of times of irradiation. Three doses of UV-C LED irradiation resulted in a 91.76 ± 3.33% death rate, as observed through SYTOX Green staining, with no rebound within 72 h. This research is the first report to observe that delayed cellular apoptosis occurred in Chlorella sp., and we expect that our study can be used as a standard reference for future industrial applications.

Mercury (Hg) is a dangerous heavy metal that can accumulate in fish and is harmful when consumed by humans. This study investigated the bioaccumulation of mercury in the form of methylmercury (MeHg) and evaluated nonspecific immune responses such as phagocytic activity and superoxide anion (O2−) production in hybrid grouper (Epinephelus fuscoguttatus × E. lanceolatus). The hybrid grouper leukocytes were incubated with methylmercury chloride (CH3HgCl) at concentrations of 10–10,000 µg/L to determine cell viability, phagocytic activity, and O2− production in vitro. Subsequently, the grouper were exposed daily to CH3HgCl mixed in the experimental diets at concentrations of 0, 1, 5, and 10 mg/kg for 28 days. The bioaccumulation of MeHg in the liver, head kidney, and muscle tissue was measured, and the phagocytic activity and O2− production were evaluated. In vitro results indicated that cell viability was significantly lower than that of the control group at concentrations > 500 µg/L. The phagocytic rate and O2− production at concentrations ˃ 500 and ˃ 200 µg/L, respectively, were significantly lower than those of the control group. The dietary exposure demonstrated that MeHg accumulated more substantially in the liver and head kidney compared with the muscle tissue in the treatment groups. Moreover, the cumulative concentration significantly increased with higher concentrations and more days of exposure. The phagocytic rate and O2− production in the treatment groups were significantly lower than those in the control group from days 2 and 1, respectively. In conclusion, hybrid grouper accumulated significant MeHg in the liver and head kidney compared with the muscle tissue, and higher concentrations and more exposure days resulted in decreased cell viability, phagocytic activity, and O2− production.

Moringa is widely known as a plant with high medicinal properties. Therefore, moringa has a high potential for use as an immunostimulant in shrimp. This study investigated the effect of a moringa water extract on the immune response, resistance against V. alginolyticus, and growth performance of whiteleg shrimp. To perform the in vitro assay, hemocytes were incubated with different concentrations of the moringa extract. Furthermore, the moringa extract was incorporated at 0 (control), 1.25 g (ME1.25), 2.5 g (ME2.5), and 5.0 g (ME5.0) per kg of diet for the in vivo assay. During the rearing period, immune responses, namely the total hemocyte count (THC), phenoloxidase (PO) activity, phagocytosis activity, superoxide anion production, and immune-related gene expression were examined on days 0, 1, 2, 4, 7, 14, 21, and 28. Growth performance was measured 60 days after the feeding period. Furthermore, the shrimp were challenged with V. alginolyticus after being fed for different feeding durations. The results of the in vitro assay revealed that 100–250 ppm of the moringa extract enhanced the PO activity, phagocytic rate (PR), and superoxide anion production. The findings of the in vivo assay demonstrated that the THC, PO activity, PR, and immune-related gene expression, including alpha-2-macroglobulin, prophenoloxidase II, penaeidin2, penaeidin3, anti-lipopolysaccharide factor, crustin, lysozyme, superoxide dismutase, and clotting protein, were higher in the group of ME.25 and ME5.0 than in the control and ME1.25 at several time points. Growth performance was significantly increased (p < 0.05) in the ME2.5 group compared to the control group. Furthermore, the dietary ME2.5 resulted in a higher survival rate compared to that of the control group after challenging with V. alginolyticus, especially at ME2.5 administered for 4 and 7 days. This study indicated that the incorporation of the moringa extract at 2.5 g per kg of diet enhanced the immune response, the growth performance of the whiteleg shrimp, and the resistance against V. alginolyticus infection.

R-phycoerythrin (R-PE), a pigment complex found in red algae, was extracted and purified from a newly identified red alga, Colaconema formosanum, and its bioactivities were examined. It was revealed that R-PE treatment resulted in high cell viability (>70%) to the mammalian cell lines NIH-3T3, RBL-2H3, RAW264.7, and Hs68, and had no effect on cell morphology in NIH-3T3 cells. Its suppression effect was insignificant on the production of IL-6 and TNF-α in lipopolysaccharides-stimulated RAW264.7 cells. However, calcium ionophore A23187-induced β-hexosaminidase release was effectively inhibited in a dose-dependent manner in RBL-2H3 cells. Additionally, it was revealed to be non-irritating to bionic epidermal tissues. Notably, procollagen production was promoted in Hs68 cells. Overall, the data revealed that R-PE purified from C. formosanum exhibits anti-allergic and anti-aging bioactivities with no observed consequential toxicity on multiple mammalian cell lines as well as epidermal tissues, suggesting that this macromolecule is a novel material for potential cosmetic use.

We investigated whether phycoerythrin (PE), a pigment sourced from marine algae, could act as an immunomodulatory agent in whiteleg shrimp (Litopenaeus vannamei). To this end, PE was extracted and purified from a PE-rich macroalgae, Colaconema sp. Our in vitro analysis demonstrated that PE enhanced prophenoloxidase and phagocytosis activity but inhibited the production of reactive oxygen species in hemocytes. Additionally, the PE signal could be detected using an in vivo imaging system after its injection into the ventral sinus of the cephalothorax of whiteleg shrimp. The expression profiles of fourteen immune-related genes were monitored in hemocytes from whiteleg shrimp injected with 0.30 μg of PE per gram of body weight, and crustin, lysozyme, penaiedin 4, and anti-lipopolysaccharide factor showed up-regulated post-stimulation. The induction of immune genes and enhancement of innate immune parameters by PE may explain the higher survival rates for shrimp that received different doses of PE prior to being challenged with Vibrio parahaemolyticus or white spot syndrome virus compared to controls. Combined, these results show that PE from Colaconema sp. can differentially stimulate the immune response of whiteleg shrimp in vitro and in vivo and could potentially be used as an immunomodulator in shrimp culture.

Plant-derived materials are believed as potential nutrient sources to be applied in aquaculture. Although many studies to assess the benefits of plant extracts have been conducted, however effects of dietary katuk (Sauropus androgynus L. Merr.) on growth performances, feeding behavior and water quality of grouper (Epinephelus coioides) are not well known. In this study, 25 grouper juveniles weighing 11.27 ± 2.53 g were reared into 100-L tank (60 x 50 x 35 cm) and cultivated for 70 days. The fish were divided into four groups in triplicate, and were offered diet without katuk extract (control), diet supplemented with 1% katuk extract (SAA); diet supplemented with 2.5% katuk extract (SAB) and diet supplemented with 5.0% katuk extract (SAC) twice daily.Statistical analyses showed that dietary katuk extract caused a significant (P < 0.05) increase in growth rate and feed intake. The efficiency of feed was also significant when fish offered dietssupplemented with 1% of katuk extract which indicated by a lower feed conversion ratio. However, no statistical differences were observed on the survival rate, condition factor, viscerosomatic indexand hepatosomatic index. Observation on feeding behavior found that all treated fish consumed compounded diet just after provided into their tank and there were no abnormal behavior or any healthy problems during experimental periods. It demonstrated that application of katuk extract in fish diets is acceptable and can stimulate the fish appetites. In conclusion, our studies indicated that dietary katuk (Sauropus androgynus) extract can be applied in aquaculture to stimulate the growth and improve feed utilization.

The health of the host is significantly influenced by the gut microbiota. Penaeus vannamei (white shrimp) is one of the most profitable aquaculture species globally. Synbiotics are typically used as a beneficial diet supplement for raising aquaculture species' growth capacities and enhancing immunity against pathogenicity. However, the effects of synbiotics on the white shrimp intestinal microbiota remain poorly understood. In the present study, we targeted the V3-V4 region of 16S rRNA genes to analyze the effects of synbiotics on white shrimp gut microbiota. Dietary synbiotics, having Lactobacillus acidophilus, and Moringa oleifera leaf extract were added to the white shrimps' feed in various proportions in the present study. In total, 490 operational taxonomic units yielding 23 phyla, 41 classes, 94 orders, 151 families, and 250 genera of microorganisms were obtained. The diet containing L. acidophilus at 1 × 107 CFU/g and M. oleifera at 2.5 g/kg led to an increase in the relative abundance of beneficial microorganisms through a significant decrease in the α diversity. Moreover, it upregulated several physiological pathways such as carbohydrate metabolism, signal transduction, lipid metabolism, nucleotide metabolism, amino acid metabolism, and environmental adaptation, which led to the upregulation of the AMPK, MAPK, P13K-Akt, lysosome, peroxisome, and ferroptosis signaling pathways; this enhanced growth and immunity in white shrimp. Whether a single species or a combination of different microorganisms improves growth and immunity remains unclear till now. Nevertheless, our results will facilitate further indepth investigation into beneficial microbial communities for upliftment of white shrimp aquaculture. [ABSTRACT FROM AUTHOR]