Optimization of Antioxidant Activity and Phenolic Compound Extraction Conditions from Red Seaweed (Laurencia obtuse)

ABSTRACT

Response surface methodology was used to investigate the effect of ultrasound-assisted extraction conditions including solvent:seaweed ratio (10:1–30:1), extraction temperature (30–50°C), and extraction time (30–60 min) on the total phenolic compounds and antioxidant activity of red seaweed (Laurencia obtuse) extracts. The optimum extraction parameters for maximum phenolic content were as follows: solvent:seaweed ratio, 30:1; extraction temperature, 50°C; and extraction time, 42.8 min. The experiment results showed that the solvent:seaweed ratio was the most significant parameter for the extraction. The optimum extraction conditions for maximum antioxidant activity were as follows: solvent:seaweed ratio, 24.3:1; extraction temperature, 45.3°C; and extraction time, 58 min. Under the above-mentioned conditions, the experimental total phenolic content and antioxidant activity value were 26.23 ± 0.75 mg GAE/g seaweed and 120.89 ± 0.81 TEAC, respectively, of seaweed tested, which are well compatible with the predicted contents.     

镉在海藻中的化学形态

利用化学试剂逐步提取法研究了紫菜、海带、裙带菜和羊栖菜中镉的存在化学形态。研究结果表明:镉以多种化学形态存在于4种海藻中,1 mol/L氯化钠提取态镉和 2% 醋酸提取态的镉所占比例较大,两者约占总镉含量的76.3%～92.9%。0.6 mol/L盐酸提取态、去离子水提取态和80%乙醇提取态镉所占比例较小,其中乙醇提取态(离子态镉)约仅占总镉含量的0.4%～9.2%,且对于4种海藻,紫菜中的离子态镉所占比例最低(均小于1%)。实验进一步证实以海藻中总镉含量作为检测标准不能准确反映海藻及其制品的食用安全性,因此亟需建立海藻中针对毒性较强的离子态镉的检测标准限量和快速有效的检测方法,以保障我国海藻产业和出口创汇经济的顺利发展。     

Biochemical and Nutritional Composition of Industrial Red Seaweed Used in Carrageenan Production

The main four seaweed species for carrageenan production used in industry are Eucheuma denticulatum, Kappaphycus alvarezii, Chondrus crispus, and Sarcothalia crispata. In addition, a fifth red seaweed, Furcellaria lumbricalis, is a source of furcellaran, which is very similar to carrageenan. In this study, the chemical composition including total lipid, fatty acid profile, tocopherol content, total protein, amino acid profile, and total phenolic content of the five semi-dried industrial red seaweeds were analyzed. The obtained results indicated that when considering the total commercial production of carrageenan, there is potential to develop a method to extract protein before the main process for carrageenan extraction. Protein was the most considerable compound in the selected seaweed, and it varied from 4% for E. denticulatum up to 28% for F. lumbricalis. In the future, this may lead to a multi-product extraction (biorefinery) approach to result in more than one product instead of the traditional single-product procedure.     

Effects of cellulase addition to canola meal in tilapia (Oreochromis niloticus L.) diets

The effects of commercial cellulase enzyme products on the nutritive value of canola meal (CM) were determined in tilapia fry. Nine diets were prepared by adding cellulase enzyme (1–5 g kg^?1) to control diet (containing no canola), low canola (41.7 g kg^?1 CM) and high canola (208.3 g kg^?1 CM). Tilapia fry was fed for 90 days, and growth, feed conversion ratio (FCR), body composition and nutrient digestibility were determined. Fish fed the diets containing the lowest levels of canola meal (41.7 g kg^?1) had similar final weight and FCR relative to the group fed the control diet, and high canola diet showed significantly lower growth than other groups (P < 0.05), irrespective of cellulase enzyme. The addition of canola meal to tilapia diets resulted in lower protein digestibility and cellulose digestibility. There were no significant differences in body compositions in all treatments. Addition of enzymes to canola meal diets had no significant effects on growth performance, body compositions and nutrient digestibility. The results of this study indicate that use in higher level of canola meal in tilapia diet was adversely affected growth parameters and FCR. Adding dietary cellulase enzyme in different rations to diets showed no significant differences in growth parameter.     

Characterization of β-N-Acetylhexosaminidase from rhizostomous jellyfish, Rhopilema asamushi, Mesogloea

β-N-Acetylhexosaminidase (EC 3.2.1.52) was purified from rhizostomous jellyfish mesogloea and characterized. Using two purification steps, this enzyme was purified up to 27.4-fold with a recovery rate of 46% compared with crude extract. The molecular weight of the enzyme was estimated to be about 136 kDa, composed of subunit molecular weights of 68 kDa. The enzyme activity was inhibited by SH-reagents, indicating that it contains a SH-group in its active site. The enzyme has a high affinity for pNPGlcNAc with Km value of 0.021 mM. The rate of hydrolysis of N-acetylchito-oligosaccharides tended to decrease with increasing degree of polymerization of the substrate. The parameters of k _cat were 92.0 s⁻¹ for pNPGlcNAc, 38.2 s⁻¹ for GlcNAc₂, 14.0 s⁻¹ for GlcNAc₃, 4.1 s⁻¹ for GlcNAc₄, 1.6 s⁻¹ for GlcNAc₅, 0.9 s⁻¹ for GlcNAc₆, respectively. These results suggest that this β-N-acetylhexosaminidase is an exo-fashion hydrolytic enzyme involved in chitin degradation. This revised version was published online in August 2006 with corrections to the Cover Date.     

Antioxidant Properties of Seven Cultivated and Natural Edible Seaweed Extracts from Taiwan

Seven species of cultivated and edible seaweeds found in Southern Taiwan—including Enteromorpha intestinalis, Monostorma nitidum, Ulva lactuca, Dictyopteris undulata, Sargassum hemiphyllum var. chinense, Laurencia intermedia, and Porphyra dentate—were used to determine their antioxidant activity by different extraction methods. All seaweeds were extracted using water at 90°C, sterilization at 121°C, homogenization, or enzymatic hydrolysis. Extracts were evaluated for their free radical-scavenging capacity using 1,1-diphenyl-2-pricrylhydrazyl (DPPH), their reducing power, and their superoxide anion scavenging activity. The DPPH-scavenging capacity of S. hemiphyllum var. chinense extracts produced by sterilization (98.59%), homogenization (94.72%), and carbohydrate-degrading enzyme treatment was higher than that of protease extracts, except for U. lactuca. The reducing power of S. hemiphyllum var. chinense water extracts and M. nitidum Viscozyme extracts was better than that of a commercial antioxidant. S. hemiphyllum var. chinense exhibited higher superoxide anion scavenging activity than the other species. The results demonstrate that different extracts of some of the seaweeds showed remarkable antioxidant activity in different assays. The results also provide important information for the marine food industry and suggest that seaweed might be a valuable source of natural antioxidants.     

Preparation of nori Pyropia yezoensis enriched with free amino acids by aging the culture with nori koji

Koji is commonly used in manufacturing Japanese fermented products and promotes enzymatic degradation. In recent years, a seaweed koji has been developed by culturing Aspergillus oryzae on nori Pyropia yezoensis. In the present study, we prepared nori koji by culturing A. oryzae on high- and low-quality noris and then mixing this preparation with additional dried nori. Suitable fermentation conditions for increasing the taste-active components of the nori mixed with nori koji were investigated. The mixture of nori and koji prepared from high-quality nori, with added water, cultured for 120 days at 10 °C, provided the greatest increase (7 times) in free amino acids. The changes in taste after culturing the nori and koji mixture were evaluated using a taste-sensing system. The sourness score of the nori and koji mixture increased significantly, but the scores for other attributes such as bitterness and umami richness did not increase after culture. The present study has demonstrated a clear increase in the free amino acid content of nori and a modification in the taste score by aging the culture with nori koji. These results will encourage the development of ‘aged seaweed,’ a novel value-added product with nutritional and taste elements modified using seaweed koji.

     

Nutrient removal efficiency of Hydropuntia cornea in an integrated closed recirculation system with pink shrimp Farfantepenaeus brasiliensis

In this study, we have tested the effect of seaweed stocking density in an experimental seaweed biofilter using the economically important red seaweed Hydropuntia cornea integrated with the cultivation of the pink shrimp Farfantepenaeus brasiliensis. Nutrient removal efficiency was evaluated in relation to seaweed stocking density (2.5, 4, 6 and 8 g fw L^?1). Total ammonia nitrogen (TAN) was the main nitrogen source excreted by F. brasiliensis, with concentrations ranging from 41.6 to 65 μM of NH₄⁺‐N. H. cornea specific growth rates ranged from 0.8 ± 0.2 to 1.4 ± 0.5% day^?1 with lowest growth rates at higher seaweed stocking density (8 g fw L^?1). Nutrient removal was positively correlated with the cultivation densities in the system. TAN removal efficiency increased from 61 to 88.5% with increasing seaweed stocking density. Changes in the chemical composition of the seaweed were analysed and correlated with nutrient enrichment from shrimp effluent. The red seaweed H. cornea can be cultured and used to remove nutrients from shrimp effluents in an integrated multi‐trophic aquaculture system applied to a closed recirculation system. Recirculation through seaweed biofilters in land‐based intensive aquaculture farms can also be a tool to increase recirculation practices and establish full recirculation aquaculture systems (RAS) with all their known associated benefits.     

Experimental evaluation of a composted seaweed extract as microalgal culture media

An organic culture media based on a composted liquid seaweed extract was developed and tested for growth of Chaetoceros muelleri. The extract was evaluated at two concentrations and then compared with two commercial media: Walne and agricultural fertilizer (AF). The concentrations of nitrogen and phosphorous differed between the two commercial media and the seaweed extract; however, growth of Chaetoceros muelleri was similar. The successful culture of Chaetoceros muelleri in composted liquid seaweed extract suggests the feasibility of and potential use for this natural, organic fertilizer in aquaculture. The seaweed extract is simple to obtain and inexpensive.     

Stimulating effects of pond water on digestive enzyme activity in the Pacific white shrimp, Litopenaeus vannamei (Boone)

Crude enzyme extracts were obtained from the digestive glands of Pacific white shrimp, Litopenaeus vannamei (Boone), reared in oligotrophic well water and eutrophic shrimp pond water to compare digestive enzyme activity between the two groups. Specific activities of serine protease, collagenase, amylase, cellulase, lipase and acid phosphatase were significantly higher (P < 0.01) in pond water‐reared shrimp (PW shrimp) than in well water reared‐shrimp (WW shrimp). For most enzymes assayed, specific activity was more than two times higher in PW shrimp, and cellulase activity was over six times higher. In contrast, chitinase activity was significantly higher (P < 0.001) in WW shrimp. Higher specific activity of most digestive enzymes in PW shrimp was probably due to natural productivity in the pond water that served as a source of organic substrates, and this increased activity may contribute to the growth‐enhancing effect of shrimp pond water.     

Use of probiotics Bacillus coagulans,Rhodopseudomonas palustris and Lactobacillus acidophilus as growth promoters in grass carp (Ctenopharyngodon idella) fingerlings

The present research evaluated the effects of three probiotics on the growth performance and intestinal digestive enzyme activity in fingerlings of grass carp, Ctenopharyngodon idella. Three treatments (T‐1, T‐2 and T‐3) were fed with diets containing different viable bacteria with a final concentration 10⁶ CFU g^?1 feed (Bacillus coagulans, Rhodopseudomonas palustris and Lactobacillus acidophilus, respectively) and the control was fed with basal diet without probiotics. All the diets supplemented with probiotics resulted in better (P < 0.05) final weight, daily weight gain and relative gain rate than control but no significant differences were observed among the treatment groups. The highest protease activity was observed in T‐1. However, T‐2 and T‐3 showed no difference (P > 0.05) in protease and cellulase activities compared with the control. The protease activity was higher in the foreintestine than in the hindintestine (P < 0.05), and the reverse was observed in cellulase activity. As for amylase activity, there was no difference between foreintestine and hindintestine. In conclusion, the three selected probiotics increased the growth performance of grass carp fingerlings. Furthermore, different digestive enzyme activity was observed in different intestine segment.     

Growth parameters of whiteleg shrimp Litopenaeus vannamei and red seaweed Gracilaria corticata in integrated culturing method under zero water exchange system

Growth parameters of whiteleg shrimp Litopenaeus vannamei and red seaweed Gracilaria corticata were measured using integrated culturing method under zero‐water exchange system in a 45‐day period. A 2 × 3 factorial design was used with two levels of shrimp stocking densities and three levels of seaweed weight densities. G. corticata was cultured on a net tied to a round polyethylene frame. Culture tanks were filled with 750‐L filtered seawater. A 40‐W compact fluorescent lamp was hung over each tank to provide adequate and sufficient light for seaweed growth. Growth parameters of shrimp and seaweed such as specific growth rate (SGR), weight gained (WG) and average daily growth (ADG) were computed based on the initial and final weight of shrimp and seaweed. The maximum and minimum SGR of L. vannamei (1.97 and 1.69%/day) were observed in treatment S₁A₃ (25 shrimp/m² and 400 g seaweed/m²) and S₂A₁ (50 shrimp/m² without seaweed) respectively. The best survival rate (94.67 ± 1.33%), WG (129.9 ± 2.9%) and feed conversion ratio (1.67 ± 0.04) were also observed in treatment S₁A₃. The SGR of G.corticata in the treatment S₁A₃ (1.97 ± 0.00%/day) was significantly higher, compared to others. Strong positive correlations were obtained between the density of G. corticata and the growth parameters of L. vannamei. The red seaweed G. corticata could boost the growth parameters, survival rate and total production of L. vannamei under zero‐water exchange system.     

Protein Hydrolysates from Shrimp (Metapenaeus dobsoni) Head Waste: Optimization of Extraction Conditions by Response Surface Methodology

Utilization of shrimp head waste from the processing industry offers a solution to environmental pollution and also produces valuable bioactive peptides. The present study was aimed to optimize the alcalase assisted extraction conditions by response surface methodology (RSM) to obtain protein hydrolysates from shrimp (Metapenaeus dobsoni) head waste with maximum antioxidant activity. Based on the models derived by RSM, the optimized conditions for maximum degree of hydrolysis (DH) were pH 8.2, temperature of 45.4°C, and enzyme/substrate (E/S) of 1.8%. Meanwhile, the optimized conditions for maximum antioxidant activity of protein hydrolysates as measured by 2, 2-diphenyl-1-picrylhydrozyl (DPPH) and ferric reducing antioxidant power (FRAP) assays were pH 8.2 and 8.1, temperatures of 49.7 and 59.4°C, and E/S of 2.1 and 2.7%, respectively. The maximum predicted values for DH, DPPH, and FRAP were 42.44%, 39.64% inhibition, and 8.3 μM Fe (II)/g wet wt. of sample, respectively. The extraction of protein hydrolysates at optimal conditions derived by these models resulted in DH, DPPH, and FRAPs of 40.31%, 38.93%, and 8.21 μM Fe (II)/g of sample, respectively, demonstrating the fitness of the models. The present study demonstrates the potential of Metapenaeus dobsoni head waste as a source of protein hydrolysates with prospective antioxidant activity.     

Carbon dioxide fixation by the seaweed Gracilaria lemaneiformis in integrated multi-trophic aquaculture with the scallop Chlamys farreri in Sanggou Bay, China

The red alga Gracilaria lemaneiformis was cultivated with the scallop Chlamys farreri in an integrated multi-trophic aquaculture (IMTA) system for 42 h at Sanggou Bay, located in north China. Variation in inorganic carbon in the IMTA system was determined. The experiment included three treatments each with three replicates and three scallop monoculture systems as controls. Scallop density (399.1 ± 7.85 g per microcosm) remained the same in all treatments while seaweed density differed. The seaweed density was set at three levels (treatments 1, 2, 3) with thallus wet weights of 125.3 ± 4.72 g, 252.3 ± 7.50 g, and 378.7 ± 6.51 g per microcosm, respectively. This produced bivalve to seaweed wet weight ratios of 1:0.31, 1:0.63, and 1:0.96 for treatments 1, 2, and 3, respectively. In control groups, continuous dissolution of carbon dioxide (CO₂) produced by scallops into the seawater not only caused an ongoing increase in partial pressure of CO₂ (pCO₂), 5.5 times higher than that of natural seawater, but also acidified seawater by 0.8 units after 42 h of culture. However, in all seaweed-scallop groups, the higher the algal density, the more CO₂ was absorbed; pCO₂ was lowest in treatment 3. The results suggest that a ratio of bivalve to seaweed less than 1:0.96 may produce an even stronger CO₂ sink. Overall, the integrated culture of seaweed and scallop could provide an efficient and environmentally friendly means to reduce CO₂ emissions from bivalve mariculture.     

海藻硫酸多糖抗氧化活性研究进展

本文就海藻硫酸多糖（SPS）的结构特征、制备,抗氧化机制及影响因素进行了较为详细的阐述。众多研究表明,SPS是从海藻中提取的一类生物活性多糖,是多糖分子链中单糖分子的部分羟基被硫酸根取代而形成的;它能有效地清除自由基,还能与金属离子鳌合,抑制脂质过氧化作用,具有显著抗氧化活性;制备粗多糖主要采用水提醇沉法,用离子交换、凝胶渗透和亲和层析法进行纯化;影响SPS抗氧化活性的因素主要有硫酸多糖的来源、多酚含量、相对分子质量大小以及硫酸基的含量与位置等。     

The potential use of Gracilaria sp. meal in diets for juvenile Penaeus monodon Fabricius

Five diets were formulated to investigate the effects of substituting wheat flour and soybean meal with various inclusion levels of seaweed (Gracilaria sp.) meal on the diet stability, performance and carcass composition of juvenile shrimp Penaeus monodon Fabricius. Gracilaria meal inclusion levels from 5% to 30% were tested against a control (0% seaweed) in isonitrogenous, isoenergetic diets fed for 60 days to quadruplicate groups of shrimp held in laboratory brackishwater recirculation system. Diets containing Gracilaria meal at up to 10% inclusion had no significant effects on diet water stability (after 12 h), shrimp performance or carcass composition compared with the control diet lacking seaweed. Diets containing 0-15%Gracilaria meal remained >88% water stable after 12 h, shrimp specific growth rates (SGRs) were 7.9-8.0% wet body weight day^-1. survival was 48-56%, food conversion ratios were 3.1-3.5, and final carcass lipid levels were 6.8-7.5%. The 30% inclusion levels of Gracilaria meal resulted in a significant deterioration in diet water stability (86% after 12 h), shrimp growth (SGR 7.3% wet body weight day^-1) and carcass lipid level (4.1%). It is suggested that with the low cost and increasing availability of Gracilaria, it may be a suitable ingredient for low-level inclusion into formulated diets for shrimp culture. The possibility of using seaweed meals for this purpose is discussed with regard to diversifying the market for cultured seaweeds and enhancing the integration of seaweed culture into coastal aquaculture systems.     

Recovery of Aliphatic Fatty Acids from Red Seaweed Champia parvula (C. Agardh) and Its Antifungal Action

The aim of the present study was to evaluate the antifungal activity of seaweed Champia parvula and identification of active compounds for this action. In this regard, C. parvula crude extracts were obtained from n-hexane, ethyl acetate, chloroform, methanol, and water used to study the antifungal action. Results showed that methanolic extract had potent action against C. albicans, C. tropicalis, T. mentagrophytes, and A.flavus, with the highest inhibition of 16.7 mm found against C. tropicalis. Minimum inhibition concentration at 12.5 and minimum fungal concentration at 25 μg/mL were found in the methanolic extract. Therefore, to identify the bioactive compound for this antifungal action in the methanolic extract, thin-layer chromatography (TLC) and reverse-phase high-performance liquid chromatography (RP-HPLC) were used. The RP-HPLC had three peaks with retention times at 3.30, 3.86, and 4.73 min, showing the presence of ester fatty acid compound. Further, these compounds were characterized by nuclear magnetic resonance (NMR), gas chromatography (GC), and liquid chromatography-mass spectrometry (LC-MS), which showed the presence of long-chain aliphatic fatty acid like eicosanoic acid with formation of oct-1-en-3-ol compounds attached.     

Influence of dietary protein on digestive enzyme activity, growth and tail muscle composition in redclaw crayfish, Cherax quadricarinatus (von Martens)

This study was conducted to evaluate the effects of dietary protein on digestive enzyme profiles, growth and tail muscle composition in the freshwater redclaw crayfish, Cherax quadricarinatus. Crayfish were fed five diets that consisted of a commercial crayfish pellet and experimental diets containing 13%, 18%, 25% or 32% crude protein (CP), for a period of 12 weeks. Analysis of digestive enzyme profiles from the midgut gland (MG) revealed a positive correlation between protease, amylase and cellulase activities and dietary protein level. For all treatments, carbohydrase activity levels (cellulase and amylase) were significantly higher than those detected for protease. As dietary protein was elevated, there was a general increase in specific growth rate (SGR), with the highest SGR (0.58 ± 0.06) values observed in crayfish fed the diet containing 25% CP. Feed conversion ratio (FCR) ranged between 5.84 and 6.97 and did not differ significantly among the treatment groups including the reference diet, with the exception of the low‐protein diet (13% CP) which showed an FCR of 9.31. Finally, regression analysis revealed a strong positive correlation between the level of dietary protein and CP content in the tail muscle (P=0.004; r²=0.99).     

Identification of gut‐associated amylase,cellulase and protease‐producing bacteria in three species of Indian major carps

Isolation and enumeration of amylase, cellulase and protease‐producing autochthonous bacteria in the proximal intestine (PI) and distal intestine (DI) of three species of Indian major carps, catla (Catla catla), mrigal (Cirrhinus mrigala) and rohu (Labeo rohita), were investigated using the conventional culture‐based technique. Population levels of amylolytic strains were the highest in the PI of catla and the lowest in the DI of rohu. The highest viable count of cellulase and protease‐producing bacteria was recorded in the DI and PI of mrigal respectively. Among the bacteria isolated, 10 strains (five from PI and five from DI) were selected as potent enzyme producers according to a quantitative enzyme assay. The chosen strains were further identified by 16S rRNA gene sequence analysis. The five strains isolated from catla showed high similarity to Citrobacter sp. clone W2, Enterobacter sp. JA24, Bacillus coagulans strain TR, uncultured bacterial clone Hel3bc04 and Bacillus cereus strain UST2006‐BC004. The four strains isolated from mrigal were most closely related to Bacillus sp. KCd2, uncultured bacterial clone Hel3bd09, B. cereus strain BU040901‐020 and Citrobacter freundii strain YRL11, while the strain isolated from rohu probably belonged to Bacillus sp. GV.     

高效液相色谱氢化物发生原子荧光（HPLC-HG-AFS)联用技术检测海藻食品中无机砷

海藻食品中无机砷的含量是水产品质量检验中主要的安全卫生指标。实验采用高效液相色谱氢化物发生原子荧光联用技术（HPLC-HG-AFS）对海藻食品中无机砷的检测进行了研究。通过实验优化了提取剂、提取时间及其仪器条件,建立了HPLC-HG-AFS联用技术检测海藻食品中无机砷含量的方法。用1.2 mol/L HCl在70 ℃水浴下提取1 h,用过氧化氢（H₂O₂）氧化;pH 6.0的15 mmol/L (NH₄)₂HPO₄溶液为流动相,HPLC-HG-AFS上机分析。样品加标量在0.10 mg/kg和1.00 mg/kg时的平均回收率均在92%以上,相对标准偏差均低于4%,方法精密度较高。研究结果表明,该方法测定海藻中的无机砷较为准确、可靠,为制定农业行业标准海藻食品中无机砷的测定,提供技术支持。