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The endo-1,4-β-mannanases (Ef-Man) gene from Eisenia fetida was determined to consist of 1131 bp and encode a 377 amino acid protein. The amino acid sequence showed similarity with the endo-1,4-β-mannanases of Daphnia pulex (62%), Cryptopygus antarcticus (64%), Crassostrea gigas (61%), Mytilus edulis (60%), and Aplysia kurodai (58%). The gene encoding mature Ef-Man was expressed in Pichia pastoris (GS115 strain). Based on SDS-PAGE analysis, the molecular mass of the purified recombinant Ef-Man (rEf-Man) was estimated to be 39 kDa. All catalytically important residues of endo-1,4-β-mannanases in the glycoside hydrolase (GH) family 5 were conserved in Ef-Man. The optimal temperature for rEf-Man was identified as 60 °C. HPLC and HPAEC analyses suggest that Ef-Man requires at least six subsites for efficient hydrolysis and is capable of performing transglycosylation reactions. The overall structure of rEf-Man is similar to those of GH5 family proteins, and tertiary structures around the active site are conserved among endo-1,4-β-mannanase families. X-ray crystallographic analysis supports the hydrolysis and transglycosylation reaction mechanism determined by HPLC and HPAEC analyses. [ABSTRACT FROM AUTHOR]

Soil environment contaminated by Escherichia coli O157:H7 which come from the waste of infected animals. Earthworms can live in the pathogens-polluted soil by their innate immunity. How the proteins of earthworms E. fetida will response to E. coli O157:H7-contaminated-soil still unclear? To identify the defense proteins under E. coli O157:H7 stress, we performed a proteomic analysis of earthworm under E. coli O157:H7 exposure through an iTRAQ technology. In total, we found 283 non-redundant proteins, including fibrinolytic protease 1, lombricine kinase, lysozyme, gelsolin, coelomic cytolytic factor-1, antimicrobial peptide lumbricin-l, lysenin, and et al. The proteins participate in metabolic processes, transcription, defense response to bacterium, translation, response to stress, and transport. The study will contribute to understand why earthworm can live in the pathogens-polluted environment. [ABSTRACT FROM AUTHOR]

Uniformly pigmented Eisenia andrei (Ea) and striped E. fetida (Ef) lumbricid earthworms are hermaphrodites capable of self-fertilization, cross-fertilization, and asymmetrical hybridization. The latter was detected by genotyping of F1 and F2 progeny of the controlled Ea+Ef pairs by species-specific sequences of maternal mitochondrial COI genes and maternal/paternal nuclear S28 rRNA genes. Among F1offspring there were self-fertilized Ea (aAA), Ef (fFF), and cross-fertilized fertile Ea-derived hybrids (aAF); the latter mated with Ea and gave new generation of Ea and hybrids, while mated with Ef gave Ea, Ef, Ea-derived hybrids and sterile Ef-derived hybrids (fFA). Coelomic fluid of Ea exhibits unique fluorescence spectra called here the M-fluorescence considered as a molecular biomarker of this species. Since similar fluorescence was detected also in some Ef (hypothetical hybrids?), the aim of present investigations was to identify the M-positive earthworms among families genotyped previously. It was assumed that factor/s responsible for metabolic pathways leading to production of undefined yet M-fluorophore might be encoded/controlled by alleles of hypothetical nuclear gene of Eisenia sp. segregating independently from species-specific S28 rRNA nuclear genes, where ‘MM’ or ‘Mm’ alleles determine M-positivity while ‘mm’ alleles determine M-negative phenotypes. Spectra of M-fluorescence were detected in all 10 Ea (aAAMM) and 19 Ea-derived hybrids (aAFMm), three of four Ef-derived hybrids (fFAMm) and one ‘atypical’ Ef (fFFMm) among 13 Ef earthworms. Among progeny of ‘atypical’ M-positive Ef (fFFMm) reappeared ‘typical’ M-negative Ef (fFFmm), confirming such hypothesis. Alternatively, the M-fluorescence might be dependent on unknown gene products of vertically-transmitted Ea-specific symbiotic bacteria sexually transferred to the Ef partner. Hypotheses of intrinsic and external origin of M-fluorescence might complement each other. The presence/absence of M-fluorophore does not correspond with body pigmentation patterns; Ef-characteristic banding appeared in posterior parts of hybrids body. In conclusion, Ea/Ef hybridization may serve for further studies on bi-directional gene flow. [ABSTRACT FROM AUTHOR]

Polychlorinated dibenzo- p -dioxins/dibenzofurans (PCDD/Fs), together with polycyclic aromatic hydrocarbons (PAHs), represent highly toxic and persistent organic environmental pollutants, especially due to their capability for bioaccumulation in fatty tissues. To observe the environmentally relevant effect of these compounds on earthworms, two soils naturally contaminated with PCDD/Fs and PAHs were used in our experiments. We focused on the role of CuZn- and Mn-superoxide dismutases. We assembled a full-length sequences of these molecules from Eisenia andrei earthworm and confirmed their activity. We demonstrated the significant reduction of CuZn-SOD on both mRNA and enzyme activity levels and increased levels of reactive oxygen species in earthworms kept in PCDD/F-polluted soil, which corresponds to the observed histopathologies of the earthworm intestinal wall and adjacent chloragogenous tissue. The results show an important role of CuZn-SOD in earthworm tissue damage caused by PCDD/Fs present in soil. We did not detect any significant changes in the mRNA expression or activity of Mn-SOD in these earthworms. In earthworms maintained in PAH-polluted soil the activity of both CuZn-SOD and Mn-SOD significantly increased. No histopathological changes were detected in these worms, however significant decrease of coelomocyte viability was observed. This reduced viability was most likely independent of oxidative stress. [ABSTRACT FROM AUTHOR]

Contamination of soils by metals and metalloids is an important environmental problem in many residential and industrial sites around the world. Lead is a common contaminant, which enters the soil through mining, industrial activities and waste disposal. A range of technologies can be used to remediate soil lead, however most remediation technologies adversely affect the environment and particularly soil biota. We have assessed the efficacy of vermiremediation (the use of earthworms for remediation) to reduce water extractable lead concentrations in soil. Earthworms were introduced to a sandy soil spiked with the common lead minerals cotunnite (PbCl 2 ), cerussite (PbCO 3 ), massicot (PbO) or galena (PbS) at 1000 mg (Pb) kg −1 . Lead concentrations in pore water extracted during the experiment were not significantly different in contaminated soil with and without worms. However, concentrations of lead in water from a deionised water extraction (washing) of contaminated soil were significantly lower in soil with earthworms than in soil without. Earthworms accumulated on average (±1 standard deviation) 276 ± 118, 235 ± 66, 241 ± 58 and 40 ± 30 mg kg −1 (dry weight of earthworms) of lead in their bodies, in PbCl 2 , PbCO 3 , PbO and PbS-dosed soils, respectively. During the experiment, earthworms lost weight in all contaminated soils, except those containing PbS. [ABSTRACT FROM AUTHOR]

Many commercially available agro and household chemicals are used as pesticides, repellents, and growth inhibitors against insect pests. The repeated uses of these chemicals against insect pests have caused the development of resistance in them; they also cause ill effects on nontarget organisms. The present study was aimed to evaluate the antifeedant, larvicidal, pupicidal, and biochemical effects of the solvent extracts of Solanum xanthocarpum against third instar larvae of Helicoverpa armigera. Hexane, chloroform and ethyl acetate extracts were subjected to phytochemical analysis. The results revealed the presence of terpenoids, flavonoid, and quinone. Maximum antifeedant activity of 72.30% was recorded in chloroform extract followed by hexane (69.02%) and ethyl acetate (57.40%) extracts against H. armigera. Chloroform extracts of S. xanthocarpum showed more than 60% larvicidal and pupicidal activity against H. armigera. The effective chloroform extract was fractionated with increasing polarity of solvent system (hexane, chloroform, and ethyl acetate extracts). Based on the TLC profile, nine major fractions were isolated. The fourth fraction showed higher antifeedant, larvicidal, and pupicidal activity against H. armigera. The effective fraction reduced the hemolymph and gut protein concentration in a concentration-dependent manner ( r 0.99). The effective fraction 4 showed 100% larvicidal activity at 500 ppm concentration with LC value of 227.95 ppm. The fourth fraction did not show any toxic symptom or mortality of earthworm. Based on these results, this effective fraction could be used in the development of a pesticide formulation to control insect. [ABSTRACT FROM AUTHOR]

In this study, the acute (72 h and 14 d) and chronic (28 d and 8 weeks) effects of cyfluthrin on earthworms were evaluated across different endpoints, which are mortality, growth, reproduction and enzyme activities. Cyfluthrin was rated as moderately toxic in 72-h filter paper test and low toxic in 14-day soil test. The exposure of earthworms to cyfluthrin-polluted soil for 8 weeks showed that growth of earthworms was inhibited by cyfluthrin, cocoon production and hatching were inhibited by 20–60 mg/kg cyfluthrin. Moreover, 28-day soil test on the responses of enzymes associated with antioxidation and detoxification showed that the activities of catalase (CAT) and glutathione S- transferase (GST) were initially increased by cyfluthrin at 5–20 mg/kg, but reduced at 30–60 mg/kg, peroxidase (POD) was increased by 26–102% by cyfluthrin in the early period, except 5 mg/kg on day 7, and ethoxyresorufin-O-deethylase (EROD) was increased by 29–335% by cyfluthrin after 3 days. Cyfluthrin degraded with a half-life of 24.8–34.8 d, showing the inconsistency between the continuous toxic responses of earthworms and degradation of cyfluthrin in soil. The variable responses of these indexes indicated that different level endpoints should be jointly considered for better evaluation of the environmental risk of contaminants in soil. [ABSTRACT FROM AUTHOR]

The dose- and time- dependent responses of cytochrome P450 (CYP) sub-enzyme activities to heavy metals in soil, and the relationships between biomarker responses and metal bioaccumulation in Eisenia fetida were evaluated. Earthworms were exposed to soils spiked with increasing doses of Cd, Cu, Pb or Zn for 21 d. Results demonstrated that EROD and CYP3A4 activities responded significantly with increasing dose and exposure duration. EROD activity significantly (P < 0.05) correlated with CYP3A4 activity exposed to Pb and Cu. The earthworm metal burdens had significant correlation with the total metal concentrations in soil (P < 0.01). The bioaccumulation factor (BAF) decreased with the increasing metal concentration in soil. The order of metal bioavailability to E. fetida was Cd > Zn > Cu > Pb. CYP3A4 activity in Pb-exposed earthworms had a significant correlation with the accumulated metal (P < 0.05). Both EROD and CYP3A4 activities in Cu-exposed worms negatively correlated with BAF (P < 0.05). Based on Discriminant Analysis (DA), CYPs activities were sensitive biomarkers of heavy metals exposure, and we also concluded that different biomarkers with multiple durations could be conducted in the eco-toxicological diagnosis of soil pollution. [ABSTRACT FROM AUTHOR]

The enantioselective degradation of fenoxaprop-ethyl in ecological indicator earthworm was studied and the main metabolites (fenoxaprop, 6-chloro-2,3-dihydrobenzoxazol-2-one, ethyl-2-(4-hydroxyphenoxy)propanoate, 2-(4-hydroxyphenoxy)propanoic acid) were also monitored on an enantiomeric level. The individual enantiomers of fenoxaprop-ethyl and its three chiral metabolites were prepared to study the acute toxicity to earthworm. Chiral analysis methods were set up based on HPLC–MS/MS with chiralpak IC chiral column. Fenoxaprop-ethyl was not found in earthworms, while the primary metabolite fenoxaprop was in relatively high levels indicating a quick hydrolysis degradation. Fenoxaprop was accumulated almost exclusively with R-enantiomer in earthworms and the bio-concentration factors of R-fenoxaprop and S-fenoxaprop were 1.39 and 0.17 respectively with the enantiomer fraction (EF) values about 0.99. The degradation of R-fenoxaprop in earthworms followed first-order kinetics with half-life of 1.82 day. The other metabolites could not be detected in earthworms. The calculated LC 50 values showed ecological indicator earthworm was more sensitive to the four metabolites than fenoxaprop-ethyl. Furthermore, earthworm was more sensitive to the R-form of the chiral metabolites than the S-form and rac-form. The results suggested metabolites and enantioselectivity should be taken into consideration to better predict the exposure concentration and apply ecological indicators in toxicological studies. [ABSTRACT FROM AUTHOR]

To investigate whether eckol, a phlorotannin compound from edible brown seaweed Eisenia bicyclis, could be responsible for the suppression of inflammatory processes, we examined the anti-inflammatory effects of eckol on Propionibacterium acnes-induced human skin keratinocytes (HaCaT) cells. P. acnes-treated HaCaT cells increased the expression of pro-inflammatory cytokines and chomokines. We also examined in vitro regulatory function of eckol on production of nitric oxide (NO), matrix metalloproteinase (MMP)−2 and MMP-9 as well as expression of inducible NO synthase, cyclooxygenase-2, and tumor necrosis factor-α. Eckol inhibited the expression or formation production of these proinflammatory mediators and cytokines in HaCaT cells. Additionally, treatment of HaCaT cells with P. acnes significantly increased protein kinase B (Akt) and nuclear factor kappa B (NF-κB) activation. Eckol inhibited P. acnes-mediated phosphorylation of Akt and activation of NF-κB in a dose-dependent manner. These results suggest that eckol could be a potential therapeutic agent to effectively treat the inflammatory skin disease induced by P. acnes. Practical Applications Eisenia bicyclis is a common perennial phaeophyceae (brown alga) and generally inhabits the region of Ulleung Island in the East sea of Korea. This edible seaweed is widely consumed in various ways such as appetizers, casseroles, muffins, pilafs, and soups. E. bicyclis also has various physiological properties such as antioxidant and anti-inflammatory activities because of their substantial quantities of phenolic compounds, constituted mainly by marine-derived polyphenols (phlorotannins). Eckol, one of phlorotannins from E. bicyclis, shows therapeutic effectiveness against viable P. acnes-induced inflammation in HaCaT cells. Thus, we suggest that eckol from E. bicyclis is a potential treatment for anti-inflammatory therapy for acne vulgaris. [ABSTRACT FROM AUTHOR]

Sewage sludge contains valuable plant nutrients, especially phosphorus. But unfortunately, it also contains pollutants which are hazardous for the environment. Phosphorus recovery from sewage sludge and its agricultural valorisation in recycling fertilisers based or containing recovered phosphate provides opportunities to minimise negative environmental effects caused by direct sludge application or conventional fertilisation. For validation, crystallised (struvite) and thermally treated phosphate recyclates (PRs) were chemically analysed, ecotoxicologically assessed and compared with a conventional phosphate fertiliser (triple superphosphate (TSP)). Three test species covering the environmental compartments water, sediment and soil were applied to evaluate the acute toxic effects of the phosphate fertiliser samples in laboratory tests ( Lemna minor, Gammarus fossarum, Eisenia fetida). The assessment and comparison showed that TSP was more toxic than the PRs at the higher tested concentrations, probably due to a higher water solubility and not to chemical composition. Higher concentrations of the crystallised PRs caused mostly a slightly higher negative effect on tested parameters of the duckweed and the freshwater amphipod than the thermally treated PRs. Agronomical relevant application amounts of all PRs and TSP (worst-case scenario) might not have an acute toxic effect on the soil invertebrates. The PRs might have minor effects on the growth of L. minor, and TSP might negatively affect the survival of the freshwater amphipods. Recovered phosphate-containing materials (PRs), in particular struvite, proved to be of high quality and low hazard in a relative risk ranking; thus, it could be one of the future alternatives of phosphorus fertilisation in agriculture. [ABSTRACT FROM AUTHOR]

Sewage sludge derived biochar (SSDB) was used as a supplementary material for municipal sewage sludge (SS) and wood chips mixtures (WC) treated by combined composting and vermicomposting. SSDB added to the mixture before composting resulted in significantly higher reproduction rate: on week 4 the number of cocoons increased by 213% when compared to the mixture with no biochar. On week 6 the average number of juveniles increased 11-fold in the mixture with biochar added before composting and 5-fold in the mixtures with biochar added after composting when compared to the mixture with no biochar. Biochar added before composting reduced bioavailability of Cd and Zn to E. fetida. The biochar-added vermicomposts showed good fertilizing properties except for elevated concentrations of Cr. The pH of all vermicomposts was in the range of 5.27–5.61. The obtained vermicomposts can be used as a growing medium for horticultural purposes or as an amendment in calcareous soils. [ABSTRACT FROM AUTHOR]

This research was conducted to develop effective and safe marine-derived antiviral compounds against norovirus. The ethyl acetate (EtOAc)-extract from Eisenia bicyclis exhibited strong antiviral activity against murine norovirus (MNV) as a norovirus surrogate. Among the phlorotannins from E. bicyclis, dieckol (DE) and phlorofucofuroeckol-A (PFF) were known to possess the strongest antibacterial activity. In this study, DE and PFF were evaluated for antiviral activity against MNV. DE and PFF exhibited strong anti-MNV activity with 50% effective concentration (EC50) of 0.9 μM. However, PFF exhibited more effective antiviral activity against MNV with higher selective index (668.87) than that of DE (550.60), due to its lower cell toxicity against RAW 264.7. This is the first report on the anti-MNV activity of phlorotannins from seaweed. The results obtained in this study suggest that the phlorotannins could be used as a potential source of natural antiviral agents. [ABSTRACT FROM AUTHOR]

Objectives Fucosterol is the primary sterol found in brown algae. Recently, considerable interest has been generated regarding fucosterol due to its potential antioxidant, anti-inflammatory and antidiabetic effects. The aim of this study was to investigate the protective effects of fucosterol on tert-butyl hydroperoxide ( t- BHP)- and tacrine-induced oxidative stress in HepG2 cells. Methods Fucosterol by itself exhibited no cytotoxicity at concentrations below 100 μ m by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. The increased intracellular reactive oxygen species ( ROS) and decreased glutathione levels observed in t- BHP- and tacrine-treated HepG2 cells were ameliorated by fucosterol pretreatment, indicating that the protective effects of fucosterol are mediated by the induction of cellular defence mechanisms against oxidative stress. Moreover, elevated alanine aminotransferase ( ALT) and aspartate aminotransferase ( AST) levels in tacrine-treated mice were significantly reduced after oral administration of fucosterol. Key findings The hepatoprotective effects of fucosterol may occur via an increase in the hepatic level of glutathione and a decrease in ROS production, thereby preventing hepatic damage and the resultant increases in ALT and AST activity. Conclusion These results suggest that fucosterol may be an effective hepatoprotective agent that could be useful for preventive therapies against oxidative stress-related hepatotoxicity. [ABSTRACT FROM AUTHOR]

Kelps are conspicuous foundation species in marine ecosystems that alter the composition of understory algal assemblages. While this may be due to changes in the competitive interactions between algal species, how kelp canopies mediate propagule supply and establishment success of understory algae is not well known. In Southern California, USA, Eisenia arborea forms dense kelp canopies in shallow subtidal environments and is associated with an understory dominated by red algal species. In canopy-free areas, however, the algal assemblage is comprised of mostly brown algal species. We used a combination of mensurative and manipulative experiments to test whether Eisenia facilitates the understory assemblage by reducing competition between these different types of algae by changes in biotic interactions and/or recruitment. Our results show Eisenia facilitates a red algal assemblage via inhibition of brown algal settlement into the canopy zone, allowing recruitment to occur by vegetative means rather than establishment of new individuals. In the canopy-free zone, however, high settlement and recruitment rates suggest competitive interactions shape the community there. These results demonstrate that foundation species alter the distribution and abundance of associated organisms by affecting not only interspecific interactions but also propagule supply and recruitment limitation. [ABSTRACT FROM AUTHOR]

Eisenia bicyclis is edible brown algae recognized as a rich source of bioactive derivatives mainly phlorotannins reported for their anti-oxidant properties. Of all phlorotannins identified so far, dieckol has shown the most potent effect in anti-inflammatory, radical scavenging and neuroprotective functions. However, whether dieckol up-regulates hemeoxygenase 1 (HO-1) and this mediates its anti-inflammatory effect in murine macrophages remains poorly understood. Dieckol (12.5–50 μM) inhibited nitric oxide production and attenuated inducible nitric oxide synthase, phospho (p)-PI-3 K, p-Akt, p-IKK-α/β, p-IκB-α and nuclear p-NF-κBp65 protein expressions, and NF-κB transcriptional activity in LPS (0.1 μg/ml) stimulated murine macrophages. On the other hand, dieckol up-regulated HO-1which partly mediated its anti-inflammatory effect in murine macrophages. Thus, dieckol appeared to be a potential therapeutic agent against inflammation through HO-1 up-regulation. [ABSTRACT FROM AUTHOR]

Highlights: [•] Structure of HMW laminaran EbL from Eisenia bicyclis was established. [•] EbL was investigated by chemical methods, NMR spectroscopy and mass spectrometry. [•] Laminaran was depolymerised by endo- and exo-glucanases from marine sources. [•] EbL and its oligomers inhibited colony formation of melanoma and colon cancer cells. [ABSTRACT FROM AUTHOR]

Highlights: [•] Fucosterol exhibited mixed-type inhibition against RLAR and HRAR activities. [•] Fucosterol showed noncompetitive-type inhibition against PTP1B activity. [•] Due to the sterol structure, fucosterol can bind tightly to AR in the docking result. [•] Fucosterol showed weak or no activity against AGE formation and α-glucosidase. [Copyright &y& Elsevier]

In this study, we determined 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging activities as well as the reducing power for screening of antioxidant activity of bieckols in Eisenia bicyclis (Kjellamn) Setchell ( E. bicyclis). The compounds 6,6′-bieckol, 6,8′-bieckol and 8,8′-bieckol are representative members of the phlorotannins family. The isolated bieckols displayed markedly strong DPPH and ABTS radical scavenging effects when compared to those of positive controls (BHA and Trolox). These bieckols were also found to have significant reducing power. The isolated bieckols (6,6′-bieckol, 6,8′-bieckol and 8,8′-bieckol) were found to effectively suppress the detrimental effects of O on type-II photosensitization. The concentrations required to exert a 50 % quenching effect on O (QC) were found to be 30.7, 35.7 and 49.4 μM for 6,6′-bieckol, 6,8′-bieckol and 8,8′-bieckol, respectively. Interestingly, all these bieckols were found to be superior to histidine (5.9 mM), a well-known O quencher. These results suggested that brown algae phlorotannins, and bieckols in particular, may play an important role in protecting marine organisms against sunlight damage by eliminating O. This knowledge may contribute to the development of natural bioactive products with potential applications in reducing photo-produced oxidative damage involving reactive oxygen species in living organisms. [ABSTRACT FROM AUTHOR]

Highlights: [•] E. bicyclis inhibit NF-κB-regulated inflammatory gene expression via ROS inhibition. [•] The CH2Cl2 fraction harboring fucosterol, exhibited strong anti-inflammatory activity. [•] Fucosterol suppressed protein levels of iNOS and COX-2 in LPS-stimulated macrophages. [•] Phlorotannins inhibited LPS-induced NO and ROS production in RAW 264.7 cells. [ABSTRACT FROM AUTHOR]

Abstract: In this study, a hydrophilic interaction chromatography (HILIC) condition was developed for the simultaneous determination of five major phlorotannins from an extract of Eisenia bicyclis (Kjellman) Setchell with good linearity (r 2 >0.999). Based on this method, the seasonal variations and extraction characteristics, in terms of total extraction yield and the content of the phlorotannins, were investigated under various extraction conditions. In results, the yields and phlorotannins were increased two-to-four times in summer (June–October) and then, were decreased to normal levels in winter (November–March). In the extraction of E. bicyclis, ethanol percentage in water, extraction time and washing time significantly affected the yield of the extract and the phlorotannins, whereas the temperature and the sample/solvent ratio impacted the extraction to a lesser degree. These results will be useful information in the application of this macroalga in the commercial areas related to nutraceuticals, pharmaceuticals, and cosmeceuticals. [Copyright &y& Elsevier]

In this study, we aimed to elucidate the antioxidant capacity of Eisenia bicyclis and evaluated its antioxidant activity using various assay systems such as 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, 2,2-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging, reducing power ability, and content of total polyphenol. Among all the performed experiments, the ethyl acetate fraction of E. bicyclis exhibited higher antioxidant activities. From this finding, isolation and purification were performed on the ethyl acetate fraction and identified dieckol and phlorofucofureoeckol-A by spectroscopic analyses including FAB-mass in the negative mode, 1H NMR, 13C NMR, 1H-1H COSY, HMQC, and HMBC spectra. Interestingly, ABTS radical scavenging activities of dieckol and phlorofucofuroeckol showed strong effects of 65.36% and 70.38% at a concentration of 50 μg/mL, respectively. DPPH radical scavenging and reducing power abilities were increased in a dose-dependent manner at various concentrations. These results suggest that dieckol and phlorofucofuroeckol-A of E. bicyclis may play an important role in protection from oxidative stress involving reactive oxygen species and may contribute to the development of new bio products, for example, a useful preservative to improve food quality and a drug for various oxidative damage-associated diseases. Practical Application: The results suggest that dieckol and phlorofucofuroeckol-A can be utilized as a natural source for potential application of antioxidant in food industry and drug for oxidative damage-associated diseases. [ABSTRACT FROM AUTHOR]

Abstract: In this study, four bioactive phloroglucinol derivates including phloroglucinol (1), eckol (2), dioxinodehydroeckol (3), and dieckol (4) were isolated from Eisenia bicyclis and characterized by nuclear magnetic resonance (NMR) spectroscopic methods. Moreover, the anti-inflammatory activity of these compounds was investigated on human hepatoma cell line HepG2 cells stimulated by lipopolysaccharide (LPS). It was demonstrated that LPS can induce the production of pro-inflammatory cytokines such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) as well as the expression of inflammatory mediators as cyclooxygenase-2 (COX-2), and inducible nitric oxide synthases (iNOS) from HepG2 cells. Among isolated compounds, compound (1) exhibited significant inhibition on LPS-stimulated inflammatory responses in HepG2 cells without any cytotoxicity. Herein, compound (1) suppresses the production of pro-inflammatory cytokines such as IL-1β, IL-6, and TNF-α and the expression of COX-2 and iNOS. Thus, these results indicated that phlorotannins isolated from E. bicyclis, especially compound (1), can be used as a beneficial source for preventing and treating inflammation response. [Copyright &y& Elsevier]

Streptomyces sp Mo endo-β-1,3-glucanase was found to have hydrolyzing activity toward curdlan and released laminarioligosaccharides selectively. The molecular weight was estimated to be 36000 Da and its N-terminal amino acid sequence was VTPPDISVTN. The optimal pH was 6 and the enzyme was found to be stable from pH 5 to 8. The optimal temperature was 60 °C and the activity was stable below 50 °C. The enzyme hydrolyzed selectively curdlan containing only β-1,3 linkages. The enzyme had 89% relative activity toward Laminaria digitata laminarin, which contains a small amount of β-1,6 linkages compared with curdlan, while Eisenia bicyclis laminarin with a higher amount of β-1,6-linkages, was not hydrolyzed. Mo enzyme adsorbed completely on curdlan powder. The enzymatic hydrolysis of curdlan powder resulted in the accumulation of laminaribiose (yield 81.7%). Trisaccharide was inevitably released from the hydrolysis of laminarioligosaccharides with 5 to 7 degrees of polymerization (DP). Although the enzyme cleaved off disaccharide (DP 2) from tetrasaccharide (DP 4), the reaction rate was lower than those of DP 5 to 7. The results indicated that the active site of Mo endo-β-1,3-glucanase can efficiently recognize glucosyl residue chain of greater than DP 5 and hydrolyzes the β-1,3 linkage between the 3rd and 4th glucosyl residue. [ABSTRACT FROM AUTHOR]

Abstract: Water-soluble polysaccharides were isolated from the brown alga Eisenia bicyclis, which was collected near the coast of the Republic of Korea. The structures of laminaran and fucoidan were investigated. Laminaran from E. bicyclis was determined to be a glucan with β-(1→6) side chains linked to a β-(1→3) backbone with relatively few branch points. Based on nuclear magnetic resonance (NMR) data, the ratio of the β-(1→3) and β-(1→6) linkages was estimated as 2.6:1. Fucoidan from E. bicyclis was found to contain 1,3-linked fucose residues, some 1,6-, 1,2,6-, 1,4,6-linked galactose residues and traces of mannose and xylose. In addition, the amount of sulfate in fucoidan was 13.2%. Those polysaccharides were non-cytotoxic to human melanoma SK-MEL-28 and colon cancer DLD-1 cells. Laminaran and fucoidan from E. bicyclis inhibited the colony formation of those cells. Therefore, they may have potential as antitumor agents. [Copyright &y& Elsevier]

In the present study, we investigated the antidiabetic potential of fucoxanthin via the inhibition of rat lens aldose reductase (RLAR), human recombinant aldose reductase (HRAR), advanced glycation end-product (AGE) formation, protein tyrosine phosphatase 1B (PTP1B), and α-glucosidase. Fucoxanthin displayed potent inhibitory activity against AGE formation and HRAR and RLAR activity. In addition, fucoxanthin showed potent inhibitory activity against PTP1B. However, it did not show α-glucosidase inhibitory activity below 200 μM. In addition, our kinetic study revealed that fucoxanthin competitively inhibited RLAR, while it showed mixed-type inhibition against PTP1B. In order to confirm enzyme inhibition, we predicted the 3D structure of PTP1B using Autodock 4.0 to simulate the binding of fucoxanthin. Docking simulation results demonstrated that three residues of PTP1B (Phe30, Phe52, and Gly183) interact with the two hydroxyl groups of fucoxanthin. In addition, the binding energy was negative (−7.66 kcal/mol), indicating that the three hydrogen bonds may stabilize the open form of the enzyme and potentiate tighter binding to the active site of PTP1B, resulting in more effective PTP1B inhibition. The results of the present study therefore clearly demonstrate the promising potential of fucoxanthin as a therapeutic intervention for the management of diabetes as well as diabetes-associated complications, which could be explored further. [ABSTRACT FROM AUTHOR]

Abstract: The phlorotannins (phloroglucinol, eckol, and dieckol) are active compounds found in Eisenia bicyclis, and have been widely investigated for their antioxidant, anti-tumor, and anti-cancer activities. In this study, we investigated the protective effects of these phlorotannins against pro-inflammatory responses in human umbilical vein endothelial cells (HUVECs) and in mice treated by high mobility group box 1 protein (HMGB1), and the signaling pathways involved. The protective activities of the phlorotannins were determined by measuring permeability, leukocyte adhesion and migration, and the activations of pro-inflammatory proteins in HMGB1-activated HUVECs. We found that the phlorotannins inhibited; lipopolysaccharide (LPS)-induced HMGB1 release, HMGB1-mediated barrier disruption, the expressions of cell adhesion molecules (CAMs), and the adhesion/transendothelial migration of leukocytes to human endothelial cells. The phlorotannins also suppressed acetic acid induced-hyperpermeability and carboxymethylcellulose-induced leukocytes migration in vivo. Further studies revealed that the hydroxyl groups on dieckol positively regulated these vascular barrier protective effects. Collectively, these results suggest that phloroglucinol, eckol, and dieckol protect vascular barrier integrity by inhibiting hyperpermeability, the expressions of CAMs, and the adhesion and migration of leukocytes, which confirms their potential usefulnesses for the treatment of vascular inflammatory diseases. [Copyright &y& Elsevier]

The article presents a study which examined the α-glucosidase inhibitory and protein tyrosine phosphate 1B (PTP1B) phlorotannins of Eisenia bicyclis and Ecklonia stolonifera. The study carried out column chromatography. Results show that E. bicyclis and E. stolonifera, and also their isolated phlorotannins, had α-glucosidase inhibitory and marked PTP1B activities.

The dietary value of juvenile stages (gametophyte and juvenile sporophyte) of the brown macroalga Eisenia bicyclis for post-larval and juvenile abalone Haliotis diversicolor of 2.0–6.5 mm in shell length (SL) was examined and compared with that of a benthic diatom, Nitzschia sp., in laboratory experiments. Most abalone actively fed on these diets, but there were large variations in the growth rate among the diets and among the growth stages of abalone. Growth rates of abalone fed on Nitzschia sp. were highly variable within each growth stage, but showed no clear differences among growth stages. In contrast, in abalone fed gametophytes or juvenile sporophytes, growth rates linearly increased as abalone grew. Growth rates of >60 μm SL/day were observed in juveniles of >3 mm SL fed gametophytes, and juveniles of >5 mm SL fed juvenile sporophytes. These results indicate that the dietary value of the juvenile stages of E. bicyclis for the abalone changes as they grow, and with growth juvenile abalone begin to efficiently utilize gametophytes and juvenile sporophytes in that order. [ABSTRACT FROM AUTHOR]

Abstract: Two types of β-1,3-glucanases, AkLam36 and AkLam33 with the molecular masses of 36kDa and 33kDa, respectively, were isolated from the digestive fluid of the common sea hare Aplysia kurodai. AkLam36 was regarded as an endolytic enzyme (EC 3.2.1.6) degrading laminarin and laminarioligosaccharides to laminaritriose, laminaribiose, and glucose, while AkLam33 was regarded as an exolytic enzyme (EC 3.2.1.58) directly producing glucose from polymer laminarin. AkLam36 showed higher activity toward β-1,3-glucans with a few β-1,6-linked glucose branches such as Laminaria digitata laminarin (LLam) than highly branched β-1,3-glucans such as Eisenia bicyclis laminarin (ELam). AkLam33 showed moderate activity toward both ELam and LLam and high activity toward smaller substrates such as laminaritetraose and laminaritriose. Although both enzymes did not degrade laminaribiose as a sole substrate, they were capable of degrading it via transglycosylation reaction with laminaritriose. The N-terminal amino-acid sequences of AkLam36 and AkLam33 indicated that both enzymes belong to the glycosyl hydrolase family 16 like other molluscan β-1,3-glucanases. [Copyright &y& Elsevier]

Abstract: The anti-allergic effect of an edible brown alga, Eisenia arborea, was investigated by determining the anti-degranulation activity on RBL cells or KU812 cells and the inhibitory effects on enzymatic activities involved in eicosanoid (leukotoriene and prostaglandin) synthesis. When the stimulated cells were treated with a M/C (methanol:chloroform=1:2, v/v) extract of the alga, the histamine release of the cells was suppressed in a dose-dependent manner, and eicosanoid release from the cells was also strongly inhibited. Furthermore, the M/C extract and six phlorotannins (eckol, 6,6′-bieckol, 6,8′-bieckol, 8,8′-bieckol, phlorofucofuroeckol-A, phlorofucofuroeckol-B) derived from the extract inhibited enzymatic activities responsible for eicosanoid synthesis. This is the first report of the enzymatic activity inhibition by the phlorotannins 6,6′-bieckol, 6,8′-bieckol and PFF-B. Since histamine and eicosanoids are known to mediate the allergic reaction, inhibition of these mediators and enzymatic activities could be responsible for the anti-allergic effect of the alga. [Copyright &y& Elsevier]

The feasibility of obtaining a similar growth response from juvenile pink abalone Haliotis corrugata at a research laboratory and a hatchery, when using natural feeds was evaluated. Four macroalgae, Egregia menziesii, Eisenia arborea, Macrocystis pyrifera, Gracilaria sp., and the surfgrass Phyllospadix torreyi were used as feeds. Response patterns of abalone were very similar at both facilities in terms of final length, weight, and survival, varying from 13.0 to 15.50 mm, 0.31 to 0.52 g, and 60.0% to 78.6%, respectively. Better growth was obtained when E. menziesii, M. pyrifera, and Gracilaria sp. were offered. Poor results were obtained with surfgrass. The feed conversion ratio was determined at the laboratory and did not vary significantly, ranging from 42.3 to 199.0; although a significant inverse correlation was observed with growth rate. Growth in length and weight and survival rates varied within 1.6–20.2 μm day−1, 0.123–1.664 mgday−1, 0.4–0.64%day−1, respectively. Mean growth rate in length (14.7μm day−1) and weight (1.18 mg day−1) at the hatchery were significantly higher than that obtained at the laboratory (9.4 μm day−1 and 0.77 mg day−1), which is most likely a consequence of more suitable water temperature at the hatchery. Mean survival rate was significantly higher at the laboratory (53.1%day−1) than at the hatchery (46.1%day−1). [ABSTRACT FROM AUTHOR]

To investigate the anti-allergic effects of the brown alga Eisenia arborea. A strain of Brown Norway rats known to strongly respond to immunoglobulin E (IgE) were used as an allergy model animal. The rats were immunized with ovalbumin by oral administration. The levels of serum IgE and histamine were suppressed in the rats fed a diet supplemented with dried E. arborea powder. As for the cytokine pattern, the interferon-γ production in the spleens and mesenteric lymph nodes (MLN) was enhanced, and the interleukin-4 (IL-4) production in the spleens and/or IL-10 production in the spleens and MLN were suppressed. These results, together with the change in the Th1/Th2 balance, indicate that the rats fed with E. arborea became more anti-allergic, suggesting that E. arborea might possess anti-allergic effects. [ABSTRACT FROM AUTHOR]

Samples of Eisenia arborea from Baja California Sur, México collected in November 2002 and March 2003 were studied. The following parameters were analyzed separately from blades and stipes: moisture, ash, mannitol, fat and crude fiber. Alginate was isolated and the following quality parameters were obtained: yield (22.4-23.3%), color (> 4) and viscosity (793- 2,210 m·Pa·s in 1% solution). No significant differences were obtained between two months, except for viscosity and crude fiber. Significant differences were obtained when comparing blades and stipes in viscosity, mannitol, ash and fat, while for yield, moisture and crude fiber no significant differences were obtained. FTIR spectroscopy showed that alginate from E. arborea contained similar amounts of mannuroic and guluronic acids (M/G ≈ 1). [ABSTRACT FROM AUTHOR]

Classic marine ecological paradigms view kelp forests as inherently temperate-boreal phenomena replaced by coral reefs in tropical waters. These paradigms hinge on the notion that tropical surface waters are too warm and nutrient-depleted to support kelp productivity and survival. We present a synthetic oceanographic and ecophysiological model that accurately identifies all known kelp populations and, by using the same criteria, predicts the existence of >23,500 km2 unexplored submerged (30- to 200-m depth) tropical kelp habitats. Predicted tropical kelp habitats were most probable in regions where bathymetry and upwelling resulted in mixed-layer shoaling above the depth of minimum annual irradiance dose for kelp survival. Using model predictions, we discovered extensive new deep-water Eisenia galapagensis populations in the Galápagos that increased in abundance with increasing depth to >60 m, complete with cold-water flora and fauna of temperate affinities. The predictability of deep-water kelp habitat and the discovery of expansive deep-water Galápagos kelp forests validate the extent of deep-water tropical kelp refugia, with potential implications for regional productivity and biodiversity, tropical food web ecology, and understanding of the resilience of tropical marine systems to climate change. [ABSTRACT FROM AUTHOR]

We screened in vitro antiviral activity against a salmonid pathogenic virus, infectious hematopoietic necrosis virus (IHNV), from the extracts of a total of 342 species of marine algae collected from the Japanese coastline. The anti-IHNV activity was found primarily in MeOH extracts, and the extract from one marine brown alga in particular, Eisenia bicyclis, showed high anti-IHNV activity. The anti-IHNV compound was isolated and purified as MC15 from the E. bicyclis extract, and the chemical structure was determined by several spectrometric analyses. The antiviral compound was proved to be a chlorophyll c2 derivative lacking the metal ion Mg2+. MC15 showed similar antiviral activity against other salmonid enveloped viruses such as Paralichthys olivaceus virus and Oncorhynchus masou virus, and stability against any pH and temperatures up to 100 °C. No cytotoxicity was observed at up to 5 µg/ml. The antiviral mechanism of MC15 appears to be direct inactivation of the viral particles. A time course study showed that the inactivation of IHNV was completed within 40 min when 200 PFU of IHNV was reacted with MC15 at 800 ng/ml. [ABSTRACT FROM AUTHOR]

The article describes the isolation, structure elucidation, and inhibitory effect of phlorofucofurorckol-B, from an Eisenia arborea, on histamine release . Eisenia arborea is an edible brown alga occasionally used as a folk medicine in gynecopathy in Japan. A new phlorotannin was segregated from the alga during the search for naturally occurring anti-allergic compounds from edible algae guided by the inhibitory effect on histamine release from rat basophile leukemia-2H3 cells.

Morphological variation is common in kelps (Phaeophyceae, Laminariales) and often results from differences in environmental conditions. Stipe hollowing, in particular, occurs in several kelp species worldwide but to date has been investigated for only a few species and primarily at local scales. Here we describe the patterns of stipe hollowing both within and among populations of Eisenia arborea across 800 km of its distribution along the west coast of North America. Our results indicate that there are large latitudinal differences among populations in both the occurrence of stipe hollowing (i.e. frequency of hollow stipes within a population) and hollowing extent (i.e. length of the hollow cavity relative to stipe length) in E. arborea sporophytes. Stipe length varied among our study locations, with the most southern population exhibiting the tallest stipes, the most northern population exhibiting the shortest stipes, and the three central populations exhibiting intermediate stipe lengths. The occurrence of stipe hollowing and hollowing extent also varied along a latitudinal gradient, with the more southern populations exhibiting both increased frequencies of hollowing and greater hollowing extents. In fact, although hollowing occurred in almost all (97%) of the sporophytes examined at the most southern location, it was absent in the sporophytes examined at the most northern location. Finally, although stipe hollowing was more common in larger stipes across all locations because of the overwhelming effect of location on the occurrence of stipe hollowing, stipe length alone is not a suitable predictor when considered across this species range. [ABSTRACT FROM AUTHOR]

Describes the structure of β-glucan oligomer from laminarin and its effect on human monocytes to inhibit the proliferation of U937 cells. Analysis of the average degree of polymerization value of the Eisenia bicyclis; Nuclear magnetic resonance findings; Inhibitory activity exhibited by the monocyte-conditioned medium precultured in the presence of a fraction of β-glucan oligomer from laminarin against the proliferation of human myeloid leukemia U937 cells.

The effects caused by suspended particles in seawater and sediment cover of the substrate on the attachment to the substrate of zoospores, and on the subsequent growth and survival of gametophytes of the brown algaEisenia bicycliswere examined in the laboratory. The attachment rate to the substrate of the zoospores was remarkably reduced by the sediment cover on the substrate, and it decreased to 3.8% of the no-sediment control by a slight sediment cover of 3.0 mg/cm2 (0.048 mm in thickness). The growth of gametophytes was not inhibited at 10 mg/cm2 or less of sediment cover, though it was stopped at 30 mg/cm2. The survival rate of gametophytes became 39.4% at 5 mg/cm2 on the substrate, and gametophytes were all blighted at 30 mg/cm2. The ratio of male to female of the surviving gametophytes became 81.5:18.5, when the sediment cover was 10 mg/cm2. The rate of total loss (TL[%]) of zoospores and gametophytes ofEisenia bicycliswas obtained from the following equation:TL = 100 (1 − exp[−0.0339 C] exp[−1.24 Q]), where,C(mg/L) andQ(mg/cm2) indicate the concentration of suspended particles in seawater and the sediment on the substrate, respectively. [ABSTRACT FROM AUTHOR]

Compares the dynamic flux to models of two flow-related morphologies of the kelp Eisenia arborea. Correlation between heat flux and pitch frequency; Comparison between the flow around kelps and that around a flat, smooth, rigid plate; Complexities of flow-related nutrient transport.

The effects of nutrient salts eluted from artificial algal reef blocks made with sea-bottom sediment (hedoro) and cement to the growth and survival rate of the gametophytes and juveniles of Eisenia bicyclis were investigated. When each sample block was submerged in filtered seawater for 28 days, the nutrient salt content from the hedoro increased with time and, in particular, the amounts of NO3 -N and NO2 -N reached a constant level on day 10. We observed that the lower the cement ratio, the higher the concentrations of eluted NH4 + -N and PO4 3 -P. The growth of gametophytes as well as juveniles was highest in the elution of the block with the lowest cement ratio of 30%. In con-trast, the survival rate of gametophytes and juveniles decreased linearly with time and the survival rate was higher for the hedoro blocks than for the block made of mortar. [ABSTRACT FROM AUTHOR]

SUMMARY:To clarify the physical properties of the rock substratum on which algal communities of Eisenia bicyclis and Ecklonia cava are found, studies were conducted on: (i) strata on the coast of the Miura Peninsula; (ii) uniaxial compression strength and erosive potential rate of the substratum; and (iii) bond strength of Eisenia bicyclis classified by location. Sea beds and rock strata lying immediately off promontories and coves with lush algal growth were identified at Bokke-zaki (Misaki Formation: Ms), Jyogashima Island (Hatsuse Formation: Ht), Ara-saki (Ms), Kawajiribana (Zushi Formation: Zs), and Chojyaga-saki (Zs). These have all been verified as belonging to the late Miocene (Miura Group). The compression strengths (kgf/cm2) of rocks at Ms, Zs and Ht were 73.0, 86.2 and 148.9, respectively. Erosive potential rates (%/h) of Ms, Zs and Ht strata were 4.47 × 10-2, 3.64 × 10-2 and 2.09 × 10-2, respectively. Bond strengths of Eisenia bicyclis to the substratum (kg) on Ms, Zs and Ht strata were 25.3, 27.4 and 23.1, respectively. Rocks of the Quaternary (Miyata Formation: My) with no algal community were found to be much more brittle than those in the Tertiary period. Conversely, concrete blocks were very rigid, but bond strength of Eisenia bicyclis was as low as 50% compared to Tertiary period rock. It can be seen from the observations above that the geological properties and the nature of the late Miocene rocks are the most favourable substratum for algal communities of Eisenia bicyclis and Ecklonia cava. [ABSTRACT FROM AUTHOR]

Discusses the result of cultures of small pieces of tissue cut from young sporophytes in Eisenia bicyclis. Approach used in the study of tissue culture and the developmental condition of callus from young sporophytes; Formation of aposporous gametophyte from dedifferetiated young sporophyte cell; Similarity of differentiation of callus cells produced from the young sporophyte explant to that of callus cells produced from the explant of natural materials of Ecklonia cava.

Cites a study on the growth and maturation of two to four year-old plants of Eisenia bicyclis from Honshu, Japan. Observation of lateral blades with sori in three to four year-old plants; Annual production of lateral blades; Estimate on the yearly loss of organic matter of the plants.

This article presents information related to tissue culture from the explant stipe of Eisenia bicyclis. A sporophyte of Eisenia bicyclis was collected at Enoshima, Kanagawa Prefecture, on July 23, 1989. Tissues were excised from the stipe. The surface was cleaned up with paper towels. The sterilization procedures of the explants for tissue cultures were the same as described in a previous report. Solid and liquid culture media were prepared using artificial seawater "Jamarin S" (Jamarin Laboratory) enriched with PESI medium. For the solid medium was used 1.5% bacto-agar in 60X10 mm Petri dishes.

The article presents comparative studies on critical light conditions for young Eisenia bicyclis and Ecklonia cava. On the community floor of Ei. bicyclis and Ec. cava, the light intensity is critical for the survival and growth of young fronds. Photosynthetic characteristics of young fronds of seaweed species under dim light condition will provide a key to understand the difference in vertical distributions of the species. In the present study, the improved differential gas-volumeter with large reaction and compensation vessels was used to measure photosynthesis and respiration in intact young fronds of comparatively large area of Ei. bicyclis and Ec. cava.

Discusses the production structure of Eisenia bicyclis and Ecklonia cava communities. Application of stratified cripping method; Presence of characteristic types of production structure in both species; Comparison on intensity and density levels of the production structure types.

Part I. Presents a study which examined the effects of size, shape and stiffness in the determination of wave forces on the stipitate kelps Eisenia arborea and Pterygophora californica. Information on the experiments conducted; Biological importance of wave action; Methodology used in this study; Results of the study; Discussion on the results.

Eisenia bicyclis (Kjellman) Setchell. A kelp bed dominated by the alga in Shizugawa Bay, Miyagi Prefecture, northeastern Japan (December 26, 2014). Endo et al. in this issue investigated combined effects of abiotic factors on juvenile sporophytes of this kelp (Photo by H. Endo). Cover picture from: [ABSTRACT FROM AUTHOR]