Toxicity of five phenolic compounds to brine shrimp Artemia sinica (Crustacea: Artemiidae)

The acute toxicity of five phenolic compounds each to 15 d old Artemia sinica was determined in this study. The brine shrimp A. sinica was hatched from the encysted dry eggs(Bohai Bay Brand) produced by Dongying Ocean Artemia Co., Ltd., China at 27℃± 1℃ in pre-filtered(through pores of 0.45 μm in diameter) and autoclaved seawater(salinity 31, pH 7.5–8.0) in a cilindroconical glass beaker(2000 mL in volume) under continuous illumination(provided by a side set 20 W fluorescent lamp) with slight aeration. Ten Artemia individuals from the same batch of the hatched were cultured in 10 mL toxicant solution prepared with seawater(salinity 31, pH 7.5–8.0) at room temperature(about 20℃) to determine 24 h, 48 h and 72 h medium lethal concentration(LC50) of 5 phenolic compounds each. It was found that the toxicity of n-heptylphenol was the highest followed by nonylphenol, t-butylphenol, 2,4-dichlorophenol and bisphenol A in order. The LC50 values of the 5 compounds were calculated with regression analysis. The real concentration(in μg- L1) of 5 phenolic compounds each in toxicant solutions was measured with GC/MS analysis. Significant loss of phenolic compounds caused by either adsorption or desorption was not found. The significant difference of LC50 values was found among the five compounds 3 exposure times each. The range between the highest no-observed-effect concentration(NOEC) and 100% death causing concentration of five phenolic compounds each was determined. The toxicity in term of 24 h LC50 value of n-HP was 9.10 times higher than that of BPA, 1.71 times higher than t-BP, 1.53 times higher than 2,4-DCP and 1.36 times higher than NP, respectively.     

Optimal salinity for dominant copepods in the East China Sea, determined using a yield density model

From 1997 to 2000, four field surveys were conducted in the East China Sea (ECS) (23°30′–33°00′N, 118°30′–128°00′E). A field data yield density model was used to determine the optimal salinities for 19 dominant copepod species to establish the relationship between surface salinities and abundance of those species. In addition, ecological groups of the copepods were classified based on optimal salinity and geographical distribution. The results indicate that the yield density model is suitable for determining the relationship between salinity and abundance. Cosmocalanus darwini, Euchaeta rimana, Pleuromamma gracilis, Rhincalanus cornutus, Scolecithrix danae and Pareucalanus attenuatus were determined as oceanic species, with optimal salinities of >34.0. They were stenohaline and mainly distributed in waters influenced by the Kuroshio or Taiwan warm current. Temora discaudata, T. stylifera and Canthocalanus pauper were nearshore species with optimal salinities of <33.0 and most abundant in coastal waters. The remaining 10 species, including Undinula vulgaris and Subeucalanus subcrassus, were offshore species, with optimal salinity ranging from 33.0–34.0. They were widely distributed in nearshore, offshore and oceanic waters but mainly in the mixed water of the ECS.     

Anti-oxidant and anti-inflammatory activities of ultrasonic-assistant extracted polyphenol-rich compounds from Sargassum muticum

Polypehnol is an important, potentially bioactive component of Sargassum muticum. In this study, ultrasonic assisted extraction of polyphenol-rich substances was performed using a 38% ethanol solution at a solid:liquid ratio of 1:30 at 68℃ for 32 min, determined by single-factor and response surface methodology(RSM) optimization. The content of polyphenol was 5.66 mg/g in the crude extract. Further extraction showed that the polyphenol mainly distributed in ethyl acetate(SKEE) and water phases(SKEW).The anti-oxidation test by electron spin resonance(ESR) spectrum showed that the SKEE had the strongest scavenging activity on DPPH(1,1-diphenyl-2-picrylhydrazyl) and alkyl radicals. SKEE was shown noncytotoxic but could inhibit the generation of cellular ROS, showing protective effects in H_2O_2 and AAPHinduced Vero cells and UV-B irradiated HaCaT cells. SKEE also significantly inhibited the release of NO of LPS-induced RAW 264.7 cells. Therefore, the polyphenol-rich extracts in ethanol and ethyl acetate showed excellent anti-oxidant and anti-inflammatory activities, which is beneficial to the development of high-value bio-substances.     

Antioxidant and antitumor potential of wild and IMTA-cultivated Osmundea pinnatifida

Osmundea pinnatifida is a red edible seaweed known as pepper dulse. O. pinnatifida was cultivated in the farm of ALGAplus(Ilhavo, Portugal). This farm is integrated with a seabream and seabass commercial aquaculture and uses the nutrient-enriched water resultant from the fish production as its cultivation medium in the integrated multi-trophic aquaculture(IMTA) manner. Wild and IMTA-cultivated samples of O. pinnatifida were screened for antioxidant and antitumor activities. The antioxidant capacity of solvent extracts from wild and IMTA cultivated samples was assessed in two methods(2,2-diphenyl-1-picrylhydrazyl(DPPH) and oxygen radical absorbance capacity(ORAC)), and their total phenolic contents(TPC) were estimated. Antitumor activity was evaluated in three different tumor cell lines(HepG-2, MCF-7, and SH-SY5Y) through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay. Among the solvents used for extraction, dichloromethane was the most effective to extract phenolic compounds and presented higher ORAC. A significant correlation was found between TPC and ORAC,which was also sustained by the principal components analysis(PCA). Dichloromethane extracts induced a cytostatic effect on MCF-7 cells and showed weak cytotoxicity to SH-SY5Y cells and weak impact on cell proliferation. Overall, there were no statistically significant differences in the biological activities shown by the wild and IMTA-cultivated samples. Hence, O. pinnatifida can be obtained in an economical and environmentally sustainable way through IMTA, maintaining bioactive properties in a high potential for further nutraceutical purposes.     

Characterization of Pythium chondricola associated with red rot disease of Pyropia yezoensis (Ueda) (Bangiales,Rhodophyta) from Lianyungang,China

Pyropia yezoensis(formerly Porphyra yezoensis) is an economically important red alga that is cultured extensively in China. The red rot disease occurs commonly during Pyropia cultivation,causing serious economic losses. An incidence of red rot disease was found in a P. yezoensis farm from mid-November to mid-December 2015 at Lianyungang, Jiangsu Province, China. Histopathological examination revealed that the naturally infected thalli were infected apparently by a pathogen, leading to red rot symptoms. The causative agent was isolated and identified as the oomycete Pythium chondricola by morphological analysis and sequence analysis of the internal transcribed spacer and cytochrome oxidase subunit 1(cox1). In artificial infection experiments on the P. yezoensis blades, the P. chondricola isolate was able to cause the same characteristic histopathology seen in natural infections. P. chondricola grew well at a wide range of temperatures in the range 8-31℃, salinities at 0-45 and pH 5-9. In an orthogonal test used to determine the effects of environmental factors(temperature, salinity, and zoospore concentration)on infection, the data revealed that temperature was the most important factor to affect red rot disease development, with the optimal conditions for disease expansion being 20℃, 35 salinity, and a zoospore concentration of 10~6 zoospores/mL. The results obtained from the present study prompted us to set up a comprehensive epidemiological study on Pyropia, which will provide support to maintain the healthy development of the Pyropia industry in China.     

Influence of temperature and salinity on germination of eelgrass (Zostera marina L.) seeds

Seagrass restoration as part of ocean ecosystem protection has been launched for many years all over the world, but intensive research on this subject in China has just begun in recent years. Seed broadcasting has been widely accepted as the most potentially useful method for seagrass restoration over large areas. We examined the influence of key environmental factors on seed germination to help promote eelgrass bed restoration. Under anoxic conditions, the influence of temperature and salinity on the germination rate of eelgrass (Zostera marina L.) seeds was examined at different combinations of four temperatures (4, 9, 14, and 24°C) and nine salinities (5 to 45, increment of 5). The effect of significant interaction of temperature and salinity on germination rate was observed (ANOVA) (P<0.001). The highest germination rate (83.3 ± 3.5)% was reached in 8 weeks at 14°C and salinity 5. Higher temperature significantly increased the germination rate at salinity 5 (P<0.001) during the whole observation period except for 24°C, while lower salinity significantly increased the germination rate at 14°C (P<0.001). Although significant interaction was found between temperature and salinity (P<0.001), the influence of salinity was stronger than that of temperature for the germination of eelgrass seeds. These results provide useful information for the propagation of artificial seedlings for seagrass restoration in China.     

Effects of salinity on embryonic development,survival, and growth of Crassostrea hongkongensis

This study examined the effects of salinity on embryonic development, survival, and growth of the Hong Kong oyster Crassostrea hongkongensis. The embryos, larvae, and juveniles of C. hongkongensis were held in artificial seawater at three different salinities (low, 15; medium, 23; and high, 30) to determine the optimum hatchery and nursery conditions for mass production of the seeds. Results showed that the percentage production of straight-hinged larvae from fertilized eggs was significantly lower at the high salinity than at the low- and medium-salinities (P < 0.05). The survival rates of larvae and juveniles differed significantly among the three salinity trials, with the highest survival rate observed at the low salinity (P < 0.05). The shell height of larvae was significantly larger at the low salinity than at the high and medium salinities from days 9 to 15 (P < 0.05), whereas that of juveniles was significantly larger at the low salinity than at the high and medium salinities on day 70 (P < 0.05). These results indicate that the larvae and juveniles of C. hongkongensis are tolerant to a wide range of salinities (15 to 30), but show better growth and survival at relatively low salinities. Thus, it is recommended to use relatively low salinities in hatchery and nursery systems for improved yields of C. hongkongensis.     

Competition Among Dinoflagellate Alexandrium Tam—arense, Raphidophyte Heterosigma Carterae and Diatom Skeletonema Costatum under Combinations ofTwo Temperatures and Five Salinities

Competition among HAB (Harmful Algal Bloom) species Dinoflagellate Alexandrium tamarense, Raphidophyte Heterosigma carterae, and Diatom Skeletonema costatum was studied in the laboratory. Experiments with these three major HAB species under combinations of different salinities (10, 18, 25, 30, 35) and temperatures (19℃, 25℃) were carried out. The results showed that S. costatum successfully competed with the other two species at salinities of 18, 25,30, and 35 at temperatures of 19℃ and 25℃. However, H. carterae showed its advantage at low salinity of 10 and became the single dominant species at salinity 10 and 25℃. A. tamarense could not compete successfully with the other two species especially at low salinities. However, it could remain at low density in the presence of higher densities of other algae.     

Effect of salinity on growth and first sexual maturity of Exopalaemon carinicauda (Holthuis, 1950)

To determine the optimal salinity for growth and first sexual maturity of ExopaIaemon carinicauda, the effects of salinity on growth and reproductive performance of early juvenile prawns of E. carinicauda were evaluated under laboratory conditions. Postlarvae from the same female broodstock were reared at six salinity levels （5, 10, 15, 20, 25, and 30） for 12 weeks. The specific growth rate （SGR） and survival rate （SR） under different salinity levels in the first 6 weeks were calculated and compared. SGR was significantly influenced by salinity. Prawns reared in salinity of 10 grew significantly faster （P〈0.05） than those reared in salinities of 5, 20, 25, and 30. However, ANOVA confirmed that there was no significant effect among the six salinity levels on SR. For the next 6 weeks, the body length （BL）, body weight （BW） at the first sexual maturity, and the age at median sexual maturity （As0） of females were measured and compared. Female prawns reared at salinity of 10 presented significantly shorter A50, but no significant differences among the six salinity levels for BL and BW at the first sexual maturity were observed. Based on the above information, the optimal salinity for growth and first sexual maturity of juvenile E. carinicauda is approximately 10.     

Glutamate dehydrogenase and Na+-K+ ATPase expression and growth response of Litopenaeus vannamei to different salinities and dietary protein levels

Improvement in the osmoregulation capacity via nutritional supplies is vitally important in shrimp aquaculture.The effects of dietary protein levels on the osmoregulation capacity of the Pacific white shrimp(L.vannamei) were investigated.This involved an examination of growth performance,glutamate dehydrogenase(GDH) and Na+-K+ ATPase mRNA expression,,and GDH activity in muscles and gills.Three experimental diets were formulated,containing 25%,40%,and 50% dietary protein,and fed to the shrimp at a salinity of 25.After 20 days,no significant difference was observed in weight gain,though GDH and Na+-K+ ATPase gene expression and GDH activity increased with higher dietary protein levels.Subsequently,shrimp fed diets with 25% and 50% dietary protein were transferred into tanks with salinities of 38 and 5,respectively,and sampled at weeks 1 and 2.Shrimp fed with 40% protein at 25 in salinity(optimal conditions) were used as a control.Regardless of the salinities,shrimp fed with 50% dietary protein had significantly higher growth performance than other diets;no significant differences were found in comparison with the control.Shrimp fed with 25% dietary protein and maintained at salinities of 38 and 5 had significantly lower weight gain values after 2 weeks.Ambient salinity change also stimulated the hepatosomatic index,which increased in the first week and then recovered to a relatively normal level,as in the control,after 2 weeks.These findings indicate that in white shrimp,the specific protein nutrient and energy demands related to ambient salinity change are associated with protein metabolism.Increased dietary protein level could improve the osmoregulation capacity of L.vannamei with more energy resources allocated to GDH activity and expression.     

Extracts of the seaweed Bifurcaria bifurcata display antifungal activity against human dermatophyte fungi

Bifurcaria bifurcata is a seaweed of the Order Fucales(Ochrophyta, Phaeophyceae) that can be found all year round along the Portuguese Atlantic shore. Although it is considered edible in some countries,its biotechnological potential has not yet been assessed in detail and here we report its antimicrobial potential against human dermatophyte fungi. Three samples were harvested from Baleal Island(Peniche, Portugal)harvested in winter(BbPe); from Aguda Beach(Arcozelo, Vila Nova de Gaia, Portugal), harvested in autumn(BbAg) and a sample obtained by culturing the Aguda sample under laboratory conditions(BbLC).A broth macrodilution assay was applied to determine the MIC(minimum inhibitory concentration) and MLC(minimum lethal concentration) of the B. bifurcata extracts. Results show that the BbPe MeOH extracts were the most effective and had higher antifungal activity against all the tested dermatophyte strains than the BbLC and BaAg extracts [Epidermophyton floccosum FF9(100-200 μg/mL MIC; 200 μg/mL MLC), Microsporum canis FF1(400 μg/mL MIC; 400 μg/mL MLC), Trichophyton mentagrophytes FF7(100 μg/mL MIC; 800 μg/mL MLC), M. gypseum CECT 2908(800 μg/mL MIC; 800 μg/mL MLC), T.mentagrophytes var. interdigitale CECT 2958(800 μg/mL MIC; 800 μg/mL MLC), T. rubrum CECT 2794(200 μg/mL MIC; 400 μg/mL MLC)]. In fact, only E. floccosum FF9 was sensitive to BbAg and BbLC MeOH extracts. To our knowledge, this is the first report of antifungal activity of B. bifurcata against human dermatophyte fungi.     

Screening of extraction methods for glycoproteins from jellyfish (Rhopilema esculentum) oral-arms by high performance liquid chromatography

In order to select an optimum extraction method for the target glycoprotein (TGP) from jellyfish (Rhopilema esculentum) oral-arms, a high performance liquid chromatography (HPLC)-assay for the determination of the TGP was developed. Purified target glycoprotein was taken as a standard glycoprotein. The results showed that the calibration curves for peak area plotted against concentration for TGP were linear (r = 0.9984, y = 4.5895x+47.601) over concentrations ranging from 50 to 400 mgL-1. The mean extraction recovery was 97.84% (CV2.60%). The fractions containing TGP were isolated from jellyfish (R. esculentum) oral-arms by four extraction methods: 1) water extraction (WE), 2) phosphate buffer solution (PBS) extraction (PE), 3) ultrasound-assisted water extraction (UA-WE), 4) ultrasound-assisted PBS extraction (UA-PE). The lyophilized extract was dissolved in Milli-Q water and analyzed directly on a short TSK-GEL G4000PWXL (7.8 mm×300 mm) column. Our results indicated that the UA-PE method was the optimum extraction method selected by HPLC.     

Salinity tolerance and growth response of juvenile <Emphasis Type="Italic">Oreochromis mossambicus</Emphasis> at different salinity levels

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Oxygen consumption and ammonia-N excretion of Meretrix meretrix in different temperature and salinity

Effects of temperatures and salinities on oxygen consumption and ammonia-N excretion rate of clamMeretrix meretrix were studied in laboratory from Oct. 2003 to Jan. 2004. Two schemes were designed in incremented temperature at 10, 15, 20, 25°C at 31.5 salinity and in incremented salinity at 16.0, 21.0, 26.0, 31.5, 36.0, and 41.0 at 20°C, all for 8–10 days. From 10 to 25°C, both respiration and excretion rate were increased. One-way ANOVA analysis demonstrated significant difference (P<0.01) in physiological parameters in this temperature range except between 15 and 20°C. The highestQ ₁₀ thermal coefficient value (12.27) was acquired between 10 and 15°C, and about 1 between 15 and 20°C, indicatingM. meretrix could well acclimate to temperature changes in this range. Salinity also had significant effects on respiration and excretion rate (P<0.05). The highest values of respiration and excretion rate ofM. meretrix were recorded at 16.0 salinity (20°C). These two physiological parameters decreased as salinity increased until reached the minimumQ ₁₀ value at 31.5 (20°C), then again, these parameters increased with increasing salinity from 31.5 to 41.0.M. meretrix can catabolize body protein to cope with osmotic pressure stress when environmental salinity is away from its optimal range. No significant difference was observed between 26.0 and 36.0 in salinity (P>0.05), suggesting that a best metabolic salinity range for this species is between 26.0 and 36.0. This work is supported by National High-Tech R & D Program of China. (863 Program) (2002AA603014).     

华贵栉孔扇贝稚贝盐度适应性的研究

通过室内实验研究了水温28.5～29.4℃、pH值7.6～8.1条件下,华贵栉孔扇贝Chlamys nobilis(Reeve)稚贝(壳高9.77±0.19mm)对不同盐度的适应性。结果表明:华贵栉孔扇贝稚贝适宜生存盐度为24.27~37.18,最适生存盐度为26.91~32.13,适宜生长盐度为24.56~37.09,最适生长盐度为26.91~29.52。在适宜生存盐度范围内,当盐度变化超出最适范围时,低盐端变化对相对成活率、壳高日增长率、体重日增长率影响比高盐端明显。     

Component Analysis and Antifungal Activity of the Compounds Extracted from Four Brown Seaweeds with Different Solvents at Different Seasons

The polar and nonpolar extracts of Colpomenia sinuosa, Padina pavonia, Cystoseira barbata and Sargassum vulgare collected during spring, summer and autumn were evaluated for their antifungal activities. The phytochemicals of unexplored seaweeds were analyzed by gas chromatography and mass spectrometric (GC/MS). The algal extracts were tested for their antifungal activities against Aspergillus niger, A. flavus, Penicillium parasiticus, Candida utilis and Fusarium solani. Phytochemicals were extracted from the four seaweeds with various solvents including methanol, ethanol, acetone, chloroform and dimethyl ether. Among the various extracts, methanolic extract showed the highest inhibition activity on all fungal species. Seasonal variation in antifungal activity was studied, while methanol extracts showed the best antifungal activity in spring. Cystoseira barbata was the most effective seaweed, having antifungal activity throughout the whole year. The UV-Vis phytochemical procedure and GC-MS analysis of the methanol extracts from tested species indicated the existence of different constituents. In conclusions, the compounds with antifungal activity were identified as indoles, terpenes, acetogenins, phenols, and volatile halogenated hydrocarbons.     

Production of DMS and DMSP in different physiological stages and salinity conditions in two marine algae

Dimethylsulfide(DMS) and dimethylsulfoniopropionate(DMSP) production by Scrippsiella trochoidea and Prorocentrum minimum was investigated to characterize the effects of physiological stage and salinity on DMS and DMSP pools of these two marine phytoplankton species.Axenic laboratory cultures of the two marine algae were tested for DMSP production and its conversion into DMS.The results demonstrated that both algal species could produce DMS,but the average concentration of DMS per cell in S.trochoidea(12.63 fmol/L) was about six times that in P.minimum(2.01 fmol/L).DMS and DMSP concentrations in algal cultures varied significantly at different growth stages,with high release during the late stationary growth phase and the senescent phase.DMS production induced by three salinities(22,28,34) showed that the DMS concentrations per cell in the two algal cultures increased with increasing salinity,which might result from intra-cellular DMSP up-regulation with the change of osmotic stress.Our study specifies the distinctive contributions of different physiological stages of marine phytoplankton on DMSP and DMS production,and clarifies the influence of salinity conditions on the release of DMS and DMSP.As S.trochoidea and P.minimum are harmful algal bloom species with high DMS production,they might play an additional significant role in the sulfur cycle when a red tide occurs.     

Influence of salinity and nitrogen content on production of dimethylsulfoniopropionate (DMSP) and dimethylsulfide (DMS) by Skeletonema costatum

The effects of changing salinity and nitrogen limitation on dimethylsulfoniopropionate(DMSP) and dimethylsulfide(DMS) concentrations were investigated in batch cultures of coastal diatom Skeletonema costatum,an ecologically important species.Changes in salinity from 20-32 caused no measurable variation in cell growth or culture yield,but increased intracellular DMSP per cell by 30%.Nitrogen limitation caused up to a two-fold increase in total DMSP per cell and up to a three-fold increase in DMS per cell.These changes in DMSP and DMS per cell in the Skeletonema costatum cultures with nitrogen limitation and changing salinity were primarily attributed to the physiological functions of DMSP as an osmolyte and an antioxidant.The data obtained in this study indicated that nitrogen limitation and salinity may play an important role in climate feedback mechanisms involving biologically derived DMS.     

COMBINED EFFECTS OF TEMPERATURE, IRRADIANCE AND SALINITY ON GROWTH OF DIATOM SKELETONEMA COSTATUM

3-factor experiment was used to study the combined effects of temperature, irradiance and salinity on the growth of an HAB species diatomSkeletonema costatum (Grev.) Cleve. The results showed that temperature (12, 19, 25, 32 °C), irradiance (0.02, 0.08, 0.3, 1.6)×10¹⁶ quanta/(s·cm²)) and salinity (10, 18, 25, 30, 35) significantly influenced the growth of this species. There were interactive effects between any two of and among all three physical factors on the growth. In the experiment, the most optimal growth condition forS. costatum was temperature of 25°C, salinity of 18–35 and irradiance of 1.6×10¹⁶ quanta/(s·cm²). The results indicatedS. costatum could divide at higher rate and were more likely to bloom under high temperature and high illumination from spring to fall. It was able to distribute widely in ocean and estuary due to its adaptation to a wide range of salinities. This study was supported by the PREPP and National Key Basic Research Project (2001CB4097) NNSFC No. 39950001, 20177023, 49576301, 49906007 and KZCX2-206.     

Antialgal and antilarval activities of bioactive compounds extracted from the marine dinoflagellate <Emphasis Type="Italic">Amphidinium carterae</Emphasis>

With the global ban on the application of organotin-based marine coatings by the International Maritime Organization, the development of environmentally friendly, low-toxic and nontoxic antifouling compounds for marine industries has become an urgent need. Marine microorganisms have been considered as a potential source of natural antifoulants. In this study, the antifouling potential of marine dinoflagellate Amphidinium carterae, the toxic and red-tide microalgae, was investigated. We performed a series of operations to extract the bioactive substances from Amphidinium carterae and tested their antialgal and antilarval activities. The crude extract of Amphidinium carterae showed significant antialgal activity and the EC₅₀ value against Skeletonema costatum was 55.4 μg mL^?1. After purification, the isolated bioactive substances (the organic extract C) exhibited much higher antialgal and antilarval activities with EC₅₀ of 12.9 μg mL^?1 against Skeletonema costatum and LC₅₀ of 15.1 μg mL^?1 against Amphibalanus amphitrite larvae. Subsequently, IR, Q-TOFMS, and GC-MS were utilized for the structural elucidation of the bioactive compounds, and a series of unsaturated and saturated 16- to 22-carbon fatty acids were detected. The data suggested the bioactive compounds isolated from Amphidinium carterae exhibited a significant inhibiting effect against the diatom Skeletonema costatum and Amphibalanus amphitrite larvae, and could be substitutes for persistent, toxic antifouling compounds.