Short-range end resection requires ATAD5-mediated PCNA unloading for faithful homologous recombination.

Homologous recombination (HR) requires bidirectional end resection initiated by a nick formed close to a DNA double-strand break (DSB), dysregulation favoring error-prone DNA end-joining pathways. Here we investigate the role of the ATAD5, a PCNA unloading protein, in short-range end resection, long-range resection not being affected by ATAD5 deficiency. Rapid PCNA loading onto DNA at DSB sites depends on the RFC PCNA loader complex and MRE11-RAD50-NBS1 nuclease complexes bound to CtIP. Based on our cytological analyses and on an in vitro system for short-range end resection, we propose that PCNA unloading by ATAD5 is required for the completion of short-range resection. Hampering PCNA unloading also leads to failure to remove the KU70/80 complex from the termini of DSBs hindering DNA repair synthesis and the completion of HR. In line with this model, ATAD5-depleted cells are defective for HR, show increased sensitivity to camptothecin, a drug forming protein-DNA adducts, and an augmented dependency on end-joining pathways. Our study highlights the importance of PCNA regulation at DSB for proper end resection and HR.

Antiviral Potential of Fucoxanthin, an Edible Carotenoid Purified from Sargassum siliquastrum, against Zika Virus.

Considering the lack of antiviral drugs worldwide, we investigated the antiviral potential of fucoxanthin, an edible carotenoid purified from Sargassum siliquastrum, against zika virus (ZIKV) infection. The antiviral activity of fucoxanthin was assessed in ZIKV-infected Vero E6 cells, and the relevant structural characteristics were confirmed using molecular docking and molecular dynamics (MD) simulation. Fucoxanthin decreased the infectious viral particles and nonstructural protein (NS)1 mRNA expression levels at concentrations of 12.5, 25, and 50 µM in ZIKV-infected cells. Fucoxanthin also decreased the increased mRNA levels of interferon-induced proteins with tetratricopeptide repeat 1 and 2 in ZIKV-infected cells. Molecular docking simulations revealed that fucoxanthin binds to three main ZIKV proteins, including the envelope protein, NS3, and RNA-dependent RNA polymerase (RdRp), with binding energies of -151.449, -303.478, and -290.919 kcal/mol, respectively. The complex of fucoxanthin with RdRp was more stable than RdRp protein alone based on MD simulation. Further, fucoxanthin bonded to the three proteins via repeated formation and disappearance of hydrogen bonds. Overall, fucoxanthin exerts antiviral potential against ZIKV by affecting its three main proteins in a concentration-dependent manner. Thus, fucoxanthin isolated from S. siliquastrum is a potential candidate for treating zika virus infections.

Antioxidant, Antiviral, and Anti-Inflammatory Activities of Lutein-Enriched Extract of Tetraselmis Species.

Microalgae are proposed to have powerful applications for human health in the pharmaceutical and food industries. Tetraselmis species (sp.), which are green microalgae, were identified as a source of broad-spectrum health-promoting biological activities. However, the bioactivity of these species has not been elucidated. We aimed to confirm the antioxidant, antiviral, and anti-inflammatory effects of Tetraselmis sp. extract (TEE). TEE showed 2,2-diphenyl-1-picryl-hydrazyl-hydrate radical and hydrogen peroxide scavenging activities and reduced plaque formation in Vero E6 cells infected with vaccinia virus. TEE treatment also significantly inhibited nitric oxide (NO) production and improved cell viability in lipopolysaccharide (LPS)-induced RAW264.7 cells. These anti-inflammatory effects were further analyzed in LPS-induced RAW 264.7 cells and the zebrafish model. Further, TEE reduced induced NO synthase expression and proinflammatory cytokine release, including tumor necrosis factor-α, interleukin-6, and interleukin-1ß, through MAPKs and NF-κB-dependent mechanisms. Further analysis revealed that TEE increased the survival rate and reduced cell death and NO production in an LPS-stimulated zebrafish model. Further, high-performance liquid chromatography revealed a strong presence of the carotenoid lutein in TEE. Overall, the results suggest that lutein-enriched TEE may be a potent antioxidant, antiviral, and anti-inflammatory agent that could be sustainably utilized in industrial applications.

Purification and Molecular Docking Study on the Angiotensin I-Converting Enzyme (ACE)-Inhibitory Peptide Isolated from Hydrolysates of the Deep-Sea Mussel Gigantidas vrijenhoeki.

The objective of this study was to prepare an angiotensin I-converting enzyme (ACE)-inhibitory peptide from the hydrothermal vent mussel, Gigantidas vrijenhoeki. The G. vrijenhoeki protein was hydrolyzed by various hydrolytic enzymes. The peptic hydrolysate exhibited the highest ACE-inhibitory activity and was fractionated into four molecular weight ranges by ultrafiltration. The <1 kDa fraction exhibited the highest ACE inhibitory activity and was found to have 11 peptide sequences. Among the analyzed peptides, KLLWNGKM exhibited stronger ACE inhibitory activity and an IC50 value of 0.007 µM. To investigate the ACE-inhibitory activity of the analyzed peptides, a molecular docking study was performed. KLLWNGKM exhibited the highest binding energy (-1317.01 kcal/mol), which was mainly attributed to the formation of hydrogen bonds with the ACE active pockets, zinc-binding motif, and zinc ion. These results indicate that G. vrijenhoeki-derived peptides can serve as nutritional and pharmacological candidates for controlling blood pressure.

Structure-Based In Silico Screening of Marine Phlorotannins for Potential Walrus Calicivirus Inhibitor.

A new calicivirus isolated from a walrus was reported in 2004. Since unknown marine mammalian zoonotic viruses could pose great risks to human health, this study aimed to develop therapeutic countermeasures to quell any potential outbreak of a pandemic caused by this virus. We first generated a 3D model of the walrus calicivirus capsid protein and identified compounds from marine natural products, especially phlorotannins, as potential walrus calicivirus inhibitors. A 3D model of the target protein was generated using homology modeling based on two publicly available template sequences. The sequence of the capsid protein exhibited 31.3% identity and 42.7% similarity with the reference templates. The accuracy and reliability of the predicted residues were validated via Ramachandran plotting. Molecular docking simulations were performed between the capsid protein 3D model and 17 phlorotannins. Among them, five phlorotannins demonstrated markedly stable docking profiles; in particular, 2,7-phloroglucinol-6,6-bieckol showed favorable structural integrity and stability during molecular dynamics simulations. The results indicate that the phlorotannins are promising walrus calicivirus inhibitors. Overall, the study findings showcase the rapid turnaround of in silico-based drug discovery approaches, providing useful insights for developing potential therapies against novel pathogenic viruses, especially when the 3D structures of the viruses remain experimentally unknown.

Angiotensin I-Converting Enzyme (ACE) Inhibition and Molecular Docking Study of Meroterpenoids Isolated from Brown Alga, Sargassum macrocarpum.

Angiotensin I-converting enzyme (ACE) is an important blood pressure regulator. In this study, we aimed to investigate the ACE-inhibitory effects of meroterpenoids isolated from the brown alga, Sargassum macrocarpum, and the molecular mechanisms underlying ACE inhibition. Four fractions of S. macrocarpum were prepared using hexane, chloroform, ethyl acetate, and water as solvents and analyzed for their potential ACE-inhibitory effects. The chloroform fraction showed the strongest ACE-inhibitory effect, with an IC50 value of 0.18 mg/mL. Three meroterpenoids, sargachromenol, 7-methyl sargachromenol, and sargaquinoic acid, were isolated from the chloroform fraction. Meroterpenoids isolated from S. macrocarpum had IC50 values of 0.44, 0.37, and 0.14 mM. The molecular docking study revealed that the ACE-inhibitory effect of the isolated meroterpenoids was mainly attributed to Zn-ion, hydrogen bonds, pi-anion, and pi-alkyl interactions between the meroterpenoids and ACE. These results suggest that S. macrocarpum could be a potential raw material for manufacturing antihypertensive nutraceutical ingredients.

ATAD5 restricts R-loop formation through PCNA unloading and RNA helicase maintenance at the replication fork.

R-loops are formed when replicative forks collide with the transcriptional machinery and can cause genomic instability. However, it is unclear how R-loops are regulated at transcription-replication conflict (TRC) sites and how replisome proteins are regulated to prevent R-loop formation or mediate R-loop tolerance. Here, we report that ATAD5, a PCNA unloader, plays dual functions to reduce R-loops both under normal and replication stress conditions. ATAD5 interacts with RNA helicases such as DDX1, DDX5, DDX21 and DHX9 and increases the abundance of these helicases at replication forks to facilitate R-loop resolution. Depletion of ATAD5 or ATAD5-interacting RNA helicases consistently increases R-loops during the S phase and reduces the replication rate, both of which are enhanced by replication stress. In addition to R-loop resolution, ATAD5 prevents the generation of new R-loops behind the replication forks by unloading PCNA which, otherwise, accumulates and persists on DNA, causing a collision with the transcription machinery. Depletion of ATAD5 reduces transcription rates due to PCNA accumulation. Consistent with the role of ATAD5 and RNA helicases in maintaining genomic integrity by regulating R-loops, the corresponding genes were mutated or downregulated in several human tumors.

In Silico Virtual Screening of Marine Aldehyde Derivatives from Seaweeds against SARS-CoV-2.

Coronavirus disease 2019, caused by the outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is an ongoing global pandemic that poses an unprecedented threat to the global economy and human health. Several potent inhibitors targeting SARS-CoV-2 have been published; however, most of them have failed in clinical trials. This study aimed to assess the therapeutic compounds among aldehyde derivatives from seaweeds as potential SARS-CoV-2 inhibitors using a computer simulation protocol. The absorption, distribution, metabolism, excretion, and toxicity (ADME/Tox) properties of the compounds were analyzed using a machine learning algorithm, and the docking simulation of these compounds to the 3C-like protease (Protein Data Bank (PDB) ID: 6LU7) was analyzed using a molecular docking protocol based on the CHARMm algorithm. These compounds exhibited good drug-like properties following the Lipinski and Veber rules. Among the marine aldehyde derivatives, 4-hydroxybenzaldehyde, 3-hydroxybenzaldehyde, 3,4-dihydroxybenzaldehyde, and 5-bromoprotocatechualdehyde were predicted to have good absorption and solubility levels and non-hepatotoxicity in the ADME/Tox prediction. 3-hydroxybenzaldehyde and 3,4-dihydroxybenzaldehyde were predicted to be non-toxic in TOPKAT prediction. In addition, 3,4-dihydroxybenzaldehyde was predicted to exhibit interactions with the 3C-like protease, with binding energies of -71.9725 kcal/mol. The computational analyses indicated that 3,4-dihydroxybenzaldehyde could be regarded as potential a SARS-CoV-2 inhibitor.

Identifying Potential Antioxidant Properties from the Viscera of Sea Snails (Turbo cornutus).

Turbo cornutus, the horned turban sea snail, is found along the intertidal and basaltic shorelines of Jeju Island, Korea. T. cornutus feeds on seaweeds (e.g., Undaria sp., and Ecklonia sp.) composed of diverse antioxidants. This study identified potential antioxidant properties from T. cornutus viscera tissues. Diverse extracts were evaluated for their hydrogen peroxide (H2O2) scavenging activities. T. cornutus viscera protamex-assisted extracts (TVP) were purified by gel filtration chromatography (GFC), and potential antioxidant properties were analyzed for their amino acid sequences and its peroxidase inhibition effects by in silico molecular docking and in vitro analysis. According to the results, T. cornutus viscera tissues are composed of many protein contents with each over 50%. Among the extracts, TVP possessed the highest H2O2 scavenging activity. In addition, TVP-GFC-3 significantly decreased intracellular reactive oxygen species (ROS) levels and increased cell viability in H2O2-treated HepG2 cells without cytotoxicity. TVP-GFC-3 comprises nine low molecular bioactive peptides (ELR, VGPQ, TDY, ALPHA, PAH, VDY, WSDK, VFSP, and FAPQY). Notably, the peptides dock to the active site of the myeloperoxidase (MPO), especially TDY and FAPQY showed the MPO inhibition effects with IC50 values of 646.0 ± 45.0 µM and 57.1 ± 17.7 µM, respectively. Altogether, our findings demonstrated that T. cornutus viscera have potential antioxidant properties that can be used as high value-added ingredients.

The Effects of Marine Algal Polyphenols, Phlorotannins, on Skeletal Muscle Growth in C2C12 Muscle Cells via Smad and IGF-1 Signaling Pathways.

Skeletal muscle is an important tissue in energy metabolism and athletic performance. The use of effective synthetic supplements and drugs to promote muscle growth is limited by various side effects. Moreover, their use is prohibited by anti-doping agencies; hence, natural alternatives are needed. Therefore, we evaluated the muscle growth effect of substances that can act like synthetic supplements from edible marine algae. First, we isolated six marine algal polyphenols belonging to the phlorotannin class, namely dieckol (DK), 2,7″-phloroglucinol-6,6'-bieckol (PHB), phlorofucofuroeckol A (PFFA), 6,6'-bieckol (6,6-BK), pyrogallol-phloroglucinol-6,6'-bieckol (PPB), and phloroglucinol (PG) from an edible brown alga, Ecklonia cava and evaluated their effects on C2C12 myoblasts proliferation and differentiation. Of the six phlorotannin isolates evaluated, DK and PHB induced the highest degree of C2C12 myoblast proliferation. In addition, DK and PHB regulates myogenesis by down-regulating the Smad signaling, a negative regulator, and up-regulating the insulin-like growth factor-1 (IGF-1) signaling, a positive regulator. Interestingly, DK and PHB bind strongly to myostatin, which is an inhibitor of myoblast proliferation, while also binding to IGF-1 receptors. Moreover, they bind to IGF-1 receptor. These results suggest that DK and PHB are potential natural muscle building supplements and could be a safer alternative to synthetic drugs.

Protective Effects of An Enzymatic Hydrolysate from Octopus ocellatus Meat against Hydrogen Peroxide-Induced Oxidative Stress in Chang Liver Cells and Zebrafish Embryo.

Octopus ocellatus, a marine cephalopod distributed in the coast of South Korea, China, Japan and tropical sea, contains high amounts of taurine. In this study, an enzymatic hydrolysate obtained from O. ocellatus meat was evaluated for its antioxidant effects using a human liver cell line and zebrafish embryo model. Enzymatic hydrolysates of the O. ocellatus meat (OOM) were prepared using six different enzymes. Among the enzymatic hydrolysates, Alcalase hydrolysate of OOM (OOMAH) showed the highest scavenging effects against 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) radicals and hydrogen peroxide (H2O2). Moreover, it showed a high oxygen radical absorbance capacity (ORAC). OOMAH treatment effectively reduced the hydroxyl radical-induced DNA damage. OOMAH reduced the production of reactive oxygen species (ROS) in H2O2-treated hepatocytes without cytotoxicity. Furthermore, OOMAH improved the survival rate and reduced the intracellular ROS levels in H2O2-treated zebrafish embryos. Compositional analysis of amino acids indicated a high content of taurine in OOMAH. Current results suggest that OOMAH possesses antioxidant bioactivities and could provide protective effects against H2O2-induced oxidative stress. Therefore, OOMAH might be used as a potential resource of functional foods.

Radio-Protective Effects of Octopus ocellatus Meat Consisted of a Plentiful Taurine Against Damages Caused by Gamma Ray Irradiation.

Gamma ray irradiation causes immune suppressive responses by inducing oxidative stress such as reduction of cell viability and damages in immune cells. In this present study, we investigated whether Octopus ocellatus meet (OM) consisted of a plentiful taurine has protective effects against damages caused by oxidative stress in murine splenocytes. First of all, we prepared the aqueous extract from OM (OMA) and identified it contained a plentiful taurine content. The result also showed that OMA exhibited the antioxidant activity by scavenging DPPH and ABTS+ radicals and hydrogen peroxide. In addition, OMA improved the cell viability without cytotoxicity in gamma ray-irradiated murine splenocytes. Moreover, OMA significantly reduced the production of reactive oxygen species (ROS) in gamma ray-irradiated splenocytes. In further study, we identified that OMA protected zebrafish embryo via improving the reduced survival rate and decreasing the formation of deformity caused by the exposure of gamma ray irradiation. Also, OMA decreased the production of NO and ROS in gamma ray-irradiated zebrafish embryos as well as the induction of cell death. In these results, this study suggests that the consumption of taurine-rich foods, such as O. ocellatus, may be useful for the useful material for the protection against oxidative stress.

Protective Effects of An Water Extracts Prepared from Loliolus beka Gray Meat Against H2O2-Induced Oxidative Stress in Chang Liver Cells and Zebrafish Embryo Model.

In this study, we first evaluated protective effects of Loliolus beka in a human liver cell line and zebrafish embryo model with its anti-oxidant activity. First, we prepared the water extract from L. beka meat (LBMW) at room temperature for 24 h and revealed it consisted of a rich taurine. LBMW exhibited the scavenging effects against 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals and hydrogen peroxide (H2O2) as well as the high value of oxygen radical absorbance capacity (ORAC). Also, the hydroxyl radical-induced DNA damage was dose-dependently reduced by the treatment of LBMW. In addition, LBMW showed no cytotoxicity and reduced the production of reactive oxygen species (ROS) in H2O2-treated hepatocytes. Moreover, LBMW regulated the expression of an anti-apoptotic molecule, Bcl-2 and the expression of pro-apoptotic molecules, Bax and PARP in H2O2-treated hepatocytes as well as the increment of antioxidant mediated-HO-1 and Nrf2 protein expression. In further study, LBMW improved the survival rate and decreased the production of ROS in H2O2-treated zebrafish embryo model. Therefore, our results suggest that Loliolus beka has protective effects against H2O2-induced oxidative stress and may be used as a potential source for functional foods.

Anti-inflammatory Effects of Galactose-Taurine Sodium Salt: A Taurine Derivate in Zebrafish In Vivo Model.

Taurine, the plentiful amino acids in mammalian cells exerts various biological activities including antioxidant and anti-inflammatory effects. Inflammation can cause several diseases such as cancer, heart disease, rheumatoid arthritis and immune system reactions. Here, we investigated anti-inflammatory effects of Galactose-Taurine sodium salt (Gal-Tau), a newly synthesized taurine derivate in LPS-stimulated zebrafish embryos in vivo model. The result showed that Gal-Tau improved the survival rate and the edema in LPS-treated zebrafish embryos. Also, Gal-Tau effectively reduced the productions of nitric oxide (NO), reactive oxygen species (ROS) and cell death induced by LPS in zebrafish embryos. In addition, Gal-Tau regulated the expression levels of inflammatory mediators such as inducible NOS (iNOS) and cycloxygenase 2 (COX-2) as well as IL-6 and TNF-α, inflammatory cytokines known as important key mediators of inflammation. Taken together, this study first indicates that Gal-Tau could be considered as an effective anti-inflammatory material with its anti-inflammatory activity.

Xylose-Taurine Reduced Suppresses the Inflammatory Responses in Lipopolysaccharide-Stimulated Raw264.7 Macrophages.

Here, the anti-inflammatory effect of Xylose-Taurine reduced (X-T-R), a taurine derivate was investigated in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. X-T-R reduced the generations of nitric oxide (NO) and prostaglandin E2 (PGE2) induced by the stimulation of LPS in RAW 264.7 by suppressing the protein expression of iNOS and COX-2 known as inflammatory mediators. Also, X-R-T reduced the expression levels of the pro-inflammatory cytokines such as interleukin (IL)-1ß, IL-6 and tumor necrosis factor (TNF-α). Moreover, X-T-R inhibited the activation of nuclear factor-κB (NF-κB) and the phosphorylation of inhibitor κB (IκB)-α. In conclusion, these results first indicate that X-T-R inhibits LPS-induced inflammation by regulating the NF-κB signal pathway in macrophages.

Protective Effects of Xylose-Taurine Reduced against Damages Caused by Oxidative Stress in Zebrafish Embryos In Vivo Model.

The zebrafish (Danio rerio) is useful and convenient vertebrate models in various studies in human disease and drug discovery. In this present study, we first evaluated whether Xylose-Taurine reduced (X-T-R), a taurine derivate protects zebrafish embryos against oxidative stress caused by AAPH (2,2'-Azobis(2-amidinopropane) dihydrochloride). First of all, we selected the concentration of X-T-R showing no toxicity in zebrafish embryos. We identified that X-T-R significantly increased the survival of zebrafish embryo reduced by treatment of AAPH. Also, X-T-R effectively inhibited the productions of reactive oxygen species (ROS) and nitric oxide (NO) as well as the formation of cell death in zebrafish embryos. Moreover, X-T-R down-regulated the expression levels of Bax, caspase-3, caspase-9 and p53 known as pro-apoptotic molecules, whereas up-regulated those of Bcl-2, an anti-apoptotic molecule in AAPH-treated zebrafish embryos. From these results, this study reveals that X-T-R, a taurine derivate might be a potential protector against various damages caused by oxidative stress.

Anti-inflammatory Effects of Galactose-Taurine Sodium Salt in LPS-Activated RAW 264.7 Cells.

In this study, we synthesized Galactose-Taurine sodium salt (G-T) as a functional food ingredient to enhance biological activities of taurine. Also, anti-inflammatory effects of G-T were investigated in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. G-T found to reduce the generations of the LPS-stimulated nitric oxide (NO) and prostaglandin E2 (PGE2) via down-regulating the expression levels of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). Also, G-T reduced the secretion of inflammatory cytokines including interleukin (IL)-1ß, IL-6 and tumor necrosis factor (TNF-α) in LPS-treated RAW 264.7 cells. Finally, we identified that G-T inhibits the activation of nuclear factor-κB (NF-κB) and the phosphorylation of inhibitor κB (IκB)-α. From these results, this study first suggests that G-T could be considered as an effective anti-inflammatory agent.

Heterologous Expression of Der Homologs in an Escherichia coli der Mutant and Their Functional Complementation.

UNLABELLED: The unique Escherichia coli GTPase Der (double Era-like GTPase), which contains tandemly repeated GTP-binding domains, has been shown to play an essential role in 50S ribosomal subunit biogenesis. The depletion of Der results in the accumulation of precursors of 50S ribosomal subunits that are structurally unstable at low Mg(2+) concentrations. Der homologs are ubiquitously found in eubacteria. Conversely, very few are conserved in eukaryotes, and none is conserved in archaea. In the present study, to verify their conserved role in bacterial 50S ribosomal subunit biogenesis, we cloned Der homologs from two gammaproteobacteria, Klebsiella pneumoniae and Salmonella enterica serovar Typhimurium; two pathogenic bacteria, Staphylococcus aureus and Neisseria gonorrhoeae; and the extremophile Deinococcus radiodurans and then evaluated whether they could functionally complement the E. coli der-null phenotype. Only K. pneumoniae and S Typhimurium Der proteins enabled the E. coli der-null strain to grow under nonpermissive conditions. Sucrose density gradient experiments revealed that the expression of K. pneumoniae and S Typhimurium Der proteins rescued the structural instability of 50S ribosomal subunits, which was caused by E. coli Der depletion. To determine what allows their complementation, we constructed Der chimeras. We found that only Der chimeras harboring both the linker and long C-terminal regions could reverse the growth defects of the der-null strain. Our findings suggest that ubiquitously conserved essential GTPase Der is involved in 50S ribosomal subunit biosynthesis in various bacteria and that the linker and C-terminal regions may participate in species-specific recognition or interaction with the 50S ribosomal subunit. IMPORTANCE: In Escherichia coli, Der (double Era-like GTPase) is an essential GTPase that is important for the production of mature 50S ribosomal subunits. However, to date, its precise role in ribosome biogenesis has not been clarified. In this study, we used five Der homologs from gammaproteobacteria, pathogenic bacteria, and an extremophile to elucidate their conserved function in 50S ribosomal subunit biogenesis. Among them, Klebsiella pneumoniae and Salmonella enterica serovar Typhimurium Der homologs implicated the participation of Der in ribosome assembly in E. coli Our results show that the linker and C-terminal regions of Der homologs are correlated with its functional complementation in E. coli der mutants, suggesting that they are involved in species-specific recognition or interaction with 50S ribosomal subunits.

A prebiotic role of Ecklonia cava improves the mortality of Edwardsiella tarda-infected zebrafish models via regulating the growth of lactic acid bacteria and pathogen bacteria.

In this study, the beneficial prebiotic roles of Ecklonia cava (E. cava, EC) were evaluated on the growth of lactic acid bacteria (LAB) and pathogen bacteria and the mortality of pathogen-bacteria infected zebrafish model. The result showed that the original E. cava (EC) led to the highest growth effects on three LABs (Lactobacillus brevis, L. brevis; Lactobacillus pentosus, L. pentosus; Lactobacillus plantarum; L. plantarum) and it was dose-dependent manners. Also, EC, its Celluclast enzymatic (ECC) and 100% ethanol extracts (ECE) showed the anti-bacterial activities on the fish pathogenic bacteria such as (Edwardsiella tarda; E. tarda, Streptococcus iniae; S. iniae, and Vibrio harveyi; V. harveyi). Interestingly, EC induced the higher production of the secondary metabolites from L. plantarum in MRS medium. The secondary metabolites produced by EC significantly inhibited the growth of pathogen bacteria. In further in vivo study, the co-treatment of EC and L. plantarum improved the growth and mortality of E. tarda-infected zebrafish as regulating the expression of inflammatory molecules such as iNOS and COX2. Taken together, our present study suggests that the EC plays an important role as a potential prebiotic and has a protective effect against the infection caused by E. tarda injection in zebrafish. Also, our conclusion from this evidence is that EC can be used and applied as a useful prebiotic.

A prebiotic effect of Ecklonia cava on the growth and mortality of olive flounder infected with pathogenic bacteria.

Olive flounder (Paralichthys olivaceus), also known as the Japanese flounder in Japan, is one of the most important commercial marine finfish species cultured in Korea and Japan. The purpose of this study was to evaluate how a species of brown algae (Ecklonia cava, E. cava) affects the growth rate of olive flounder and its immune response to pathogenic bacteria. First, the experimental fish were divided into four groups: the control group was fed the diet containing only 1.0% Lactobacillus plantarum (L. plantarum), group I was fed 1.0% L. plantarum and 1.0% E. cava (EC), group II was fed 1.0% L. plantarum and 0.1% ethanol extract of EC (EE), and group III was fed 1.0% L. plantarum and 0.5% EE. The diets fed to the fish twice a day for 16 weeks. The results indicated that supplementation with 1.0% EC and 0.1% EE improved the growth and body weight of olive flounder, and decreased its mortality. This diet, however, did not significantly affect the biochemical profiles of the experimental flounder. The supplementation of 1.0% EC also enhanced the innate immune response of the fish, as evidenced by the high respiratory burst, and increased serum lysozyme and myeloperoxidase activity. The addition of 1.0% EC and either 0.1% or 0.5% EE also decreased the accumulative mortality of olive flounder infected by pathogenic bacteria (Edwardsiella tarda, Streptococcus iniae, and Vibrio harveyi). Overall, these results suggest that E. cava can act as a prebiotic by improving the innate immune response in fish infected with pathogenic bacteria as increased the growth of the probiotic.