Metabolism of polychaete Neanthes japonica Izuka: relations to temperature, salinity and body weight

Polychaete Neanthes japonica is a species geographically specific in China and Japan with important scientific implication and commercial value. In this study, the relations of body weight, salinity and temperature to oxygen consumption and ammonia excretion of N. japonica were determined. Three different groups in body weight (large: 2.34 +/- 0.36 g, middle: 1.50 +/- 0.21 g and small: 0.62 +/- 0.12 g) were set for all experiments. Results show that the body weight is negatively related to the rates of oxygen consumption and ammonia excretion; and the relationship is significant. The oxygen consumption and ammonia excretion at 24A degrees C decreased at salinity from 5 to 30 and increased above 30, indicating that both lower and higher salinity are adverse and certain degree of salinity stress is necessary for enhancing the energy demand. At salinity 30, rising temperature from 18A degrees C to 30A degrees C, the oxygen consumption increased before 27A degrees C and then decreased. However, the relation of ammonia excretion and temperature seems more complex. Two-way ANOVA shows that salinity, temperature and body weight all have a significant effect on the oxygen consumption and ammonia excretion of the worm. Moreover, interaction between salinity/temperature and body weight is also significant. O:N (oxygen/nitrogen) ratio varies greatly in this case from 5.97 to 463.22, indicating that N. japonica can regulate the type of metabolic substrate against environment changes.

Observations on the nocturnal activity and feeding behavior of Anguilla japonica glass eels under laboratory conditions

Glass eels of the temperate anguillid species, Anguilla japonica, clearly showed a nocturnal activity rhythm under laboratory conditions. Light-dark cycle was a determinant factor affecting their photonegative behavior, nocturnal locomotor activity, and feeding behavior. Under natural light conditions, glass eels remained in shelters with little daytime feeding, but came out to forage during darkness. They moved and foraged actively in the following dark, and then their activity gradually declined possibly because of food satiation. They finally buried in the sand or stayed in tubes immediately after the lights came on. Under constant light, glass eels often came out of the shelters to forage in the lights but spent little time moving outside the shelters (e.g. swimming or crawling on the sand). Glass eels took shelter to avoid light and preferred tubes to sand for shelter possibly because tubes were much easier for them to take refuge in than sand. Feeding and locomotor activities of the glass eels were nocturnal and well synchronized. They appeared to depend on olfaction rather than vision to detect and capture prey in darkness. Feeding was the driving force for glass eels to come out of sand under constant light. However, in the dark, some glass eels swam or crept actively on sand even when they were fully fed. The lunar cycles of activity rhythms of glass eels that have been observed in some estuarine areas were not detected under these laboratory conditions.

Locomotor activity rhythm in the Japanese eel Anguilla japonica elvers

Under artificial LD cycles (6, 12, 18 L), the elvers of Japanese eel, Anguilla japonica, showed a 24 h cycle of locomotor activity rhythm being most active at light transitions: the eels' activity rose to a primary peak after lights-off, followed by a quiescent period during which they buried into the shelters or lying motionlessly on sand for most of the time, and then reached a secondary peak before lights-on. Elvers could resynchronize their activity rhythm with a new photo cycle within 4 d. Moreover, their activity level at dark phase significantly increased as the light period was prolonged: higher activity levels during shorter dark period. However, the elvers did not display clearly the existence of a circadian rhythm under constant light or dark conditions. The timing of daily activity rhythm evidenced in the Japanese eels may occur through the action of the LD cycles with a weak participation of an endogenous circadian system. In all the LD cycles, over 99% of the activity occurred in the dark phase, indicating that the eels were always nocturnally active no matter what time of day it might be. Under 12 L conditions, the eels' activity level and the time outside sand were significantly elevated both at light and dark phases as temperature increased from 10 similar to 15 to 20 similar to 25 degrees C. The activity rhythm pattern (i.e., two peaks occurring around light transitions) did not apparently change among temperatures. However, in contrast with the primary activity peaks immediately after lights-off at 20 and 25 degrees C, the timing of the primary peaks at 10 and 15 degrees C showed a latency of a few hours following lights-off, indicating the inhibiting effect of low temperature on the eels' activity.

Observations on the spawning behavior of artificially matured Japanese eels Anguilla japonica in captivity

Spawning behavior of artificially matured Japanese eels Anguillo japonica in captivity was investigated using a DVD Video image system. Following a routine hormone treatment technique for this fish, female eels were artificially matured by weekly intramuscular injections of salmon pituitary extracts (SPE) at a dosage of 40 mg kg(-1) BW for a total of 7-11 doses to induce ovarian maturation, while male eels received weekly intramuscular injections of human chorionic gonadotropin (HCG) at a dosage of 1000 IU kg(-1) BW for a total of 6-11 doses at 18 degrees C to induce testicular maturation in a separate aquarium. In this experiment, three pairs of such hormone-treated matured eels were acclimatized in seawater in 1.5 m(3) experimental aquaria with or without shelters at 20 degrees C for 24 h. Twenty four hours after the acclimatization terminated, the females received SPE injections to boost maturation and ovulation. Twenty four hours following these injections, the females received injections of HCG (1000 IU per fish, HCG injection) and 17 alpha-hydroxyprogesterone (2 mg per fish) to induce ovulation, while males were given HCG injections (1000 IU per fish, HCG injection) to induce spermiation. Video taping started after the 24 h acclimatization terminated and last for a total of 96 h. Before the HCG injections, both sexes were inactive, staying on the bottom or in shelters if available. Following these HCG injections, they became active and frequently left the bottom swimming in the water column. During the 24 h following HCG injections, activity accounted for 67% and 45% of the total activity in no shelter treatment for females and males, respectively, in comparison with 77% and 78% in shelter treatment. Activity was significantly more pronounced during this phase than during other phases for each sex in either shelter treatment. Egg release and sperm ejection occurred in the water column around the time eels' activity reached peaks. Eels either returned into the shelters or stayed motionlessly on the bottom of the aquaria after egg release and sperm ejection. Eight out of nine (89%) females in no shelter treatment spontaneously released eggs with a total of 11 batches 14-18 h following HCG injections, in contrast with four out of nine (44%) females releasing eggs for 4 batches 16-20 h in shelter treatment. Males arrived at activity peaks 11-13 h following HCG injections in no shelter treatment, 2-4 h ahead of the females (14-16 h), in comparison with 8-11 h in shelter treatment with 5-6 h ahead of the females (14-17 h). Courtship behavior indicative of spawning such as pairing, chasing and touching bodies was not observed in the eels in this study. However, on many occasions, eels of both sexes (male-female or female-female) were found to "cruise together" in water column for a short time period or frequently come together prior to releasing eggs and ejecting sperm, suggesting the possibility of group mating in artificially matured Japanese eels. (c) 2007 Elsevier B.V. All rights reserved.

Temperature influence on the spawning performance of artificially-matured Japanese eel, Anguilla japonica, in captivity

We studied the influence of temperature on the spawning performance of artificially matured Japanese eels, Anguilla japonica, in captivity. We used routine hormone injections to bring females and males to maturity in separate aquaria. We recorded the behavior of three pairs of such hormone-treated matured eels in an aquarium (2 replicates) at four temperatures: 14, 18, 22, and 27 degrees C, respectively. They became active and frequently left the bottom swimming in the water column, and spawning events occurred. Females released eggs in the water column around the activity peaks. Males preceded females in reaching activity peaks (presumably the timing of sperm ejection and egg release), possibly resulting in the low fertilization we observed in this experiment. Males and females returned back to the aquarium bottoms and became quiet after spawning. On several occasions, male-female or female-female pairs were observed to 'cruise together' in the water column for several to tens of seconds prior to egg releasing, but no courtship behavior indicative of spawning such as pairing and chasing was observed in the eels in our study. Our results suggest that 18-22 degrees C might be the thermal preference for spawning for Japanese eels, which approximates the temperature range of the 500 m deep water layer around the Mariana Islands seamount area, the presumed spawning site for the Japanese eel.

Effect of alginic acid decomposing bacterium on the growth of Laminaria japonica (Phaeophyceae)

We collected the diseased blades of Laminaria japonica from Yantai Sea Farm from October to December 2002, and the alginic acid decomposing bacterium on the diseased blade was isolated and purified, and was identified as Alterornonas espejiana. This bacterium was applied as the causative pathogen to infect the blades of L. japonica under laboratory conditions. The aim of the present study was to identify the effects of the bacterium on the growth of L. japonica, and to find the possibly effective mechanism. Results showed that: (1) The blades of L. japonica exhibited symptoms of lesion, bleaching and deterioration when infected by the bacterium, and their growth and photosynthesis were dramatically suppressed. At the same time, the reactive oxygen species (ROS) generation enhanced obviously, and the relative membrane permeability increased significantly. The contents of malonaldehyde (MDA) and free fatty acid in the microsomol membrane greatly elevated, but the phospholipid content decreased. Result suggested an obvious peroxidation and deesterrification in the blades of L. japonica when infected by the bacterium. (2) The simultaneous assay on the antioxidant enzyme activities demonstrated that superoxide dismutase (SOD) and catalase (CAT) increased greatly when infected by the bacterium, but glutathione peroxidase (Gpx) and ascorbate peroxidase (APX) did not exhibit active responses to the bacterium throughout the experiment. (3) The histomorphological observations gave a distinctive evidence of the severity of the lesions as well as the relative abundance in the bacterial population on the blades after infection. The bacterium firstly invaded into the endodermis of L. japonica and gathered around there, and then resulted in the membrane damage, cells corruption and ultimately, the death of L. japonica.

Synthesized different derivatives of low molecular fucoidan extracted from Laminaria japonica and their potential antioxidant activity in vitro

Six low molecular fucoidan (DFPS) derivatives were synthesized successfully, and their potential antioxidant activities were investigated employing various established in vitro systems. All DFPS derivatives possessed considerable antioxidant activity, and had stronger antioxidant ability than DFPS in certain tests. The benzoylated DFPS (PHDF) showed strongest scavenging activity on superoxide, hydroxyl and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, however, DFPS exhibited greatest reducing power. Available data suggested that substituted groups of DFPS played an important role on antioxidant activity, and the mechanism on influence the antioxidant activity of samples of substituted group was indicated. (C) 2009 Elsevier B.V. All rights reserved.

Synthesized oversulphated, acetylated and benzoylated derivatives of fucoidan extracted from Laminaria japonica and their potential antioxidant activity in vitro

Three sulphated polysaccharide derivatives (oversulphated, acetylated and benzoylated fucoidan) were successfully synthesized, and their antioxidant activities were investigated employing various established in vitro systems. A novel catalyst N-bromosuccinimide (NBS) was used in acetylation and benzoylation reaction, and the degree of acetylation was evaluated using IR and NMR spectra. Both fucoidan derivatives possessed considerable antioxidant activity, and had stronger antioxidant ability than fucoidan in certain tests. The benzoylated fucoidan showed strongest superoxide and hydroxyl radical scavenging activity, however, the acetylated fucoidan exhibited strongest activity on scavenging DPPH radical and reducing power. Available data obtained with in vitro models suggested that substituted groups of fucoidan played an important role on antioxidant activity. The mechanism on the antioxidant activity of sulphonyl, acetyl and benzoyl group is different. (C) 2008 Elsevier Ltd. All rights reserved.

Toxicological evaluation of fucoidan extracted from Laminaria japonica in Wistar rats

Fucoidans, the sulfated polysacchrides extracted from brown algae, have been extensively studied for their diverse biological activities. However, there is no detailed report investigating the toxicity of fucoidan. In this study, the acute and subchronic (6 months) toxicity of varying levels of fucoidan extracted from Laminaria japonica was investigated in Wistar rats after oral administration. The results showed that no significant toxicological changes were observed when 300 mg/kg body weight per day fucoidan was administered to rats. But when the dose was increased to 900 and 2500 mg/kg body weight per day, the clotting time was significantly prolonged. Besides this, no other signs of toxicity were observed. Based on these results, it can be concluded that the no adverse effect level of fucoidan from L. japonica is 300 mg/kg body weight per day. (C) 2004 Elsevier Ltd. All rights reserved.

Antioxidant activity of sulfated polysaccharide fractions extracted from Laminaria japonica

Fucoidan, a group of sulfated heteropolysaccharide, was extracted from Laminaria japonica, an important economic alga species in China. Three sulfated polysaccharide fractions (F1, F2, and F3) were successfully isolated through anion-exchange column chromatography and had their antioxidant activities investigated employing various established in vitro systems, including superoxide and hydroxyl radical scavenging activity, chelating ability, and reducing power. Chemical analysis suggested that F1 and F3 were heteropolysaccharide in which galactose was the major component, while F2 was a typical fucoidan. All fractions possessed considerable antioxidant activity, and F1, F2 and F3 had stronger antioxidant ability than fucoidan in certain tests. The correlation between the sulfate content and scavenging superoxide radical ability was positive. Available data obtained with in vitro models suggested that the ratio of sulfate content/fucose was an effective indicator to antioxidant activity of the samples. (C) 2007 Elsevier B.V. All rights reserved.

The extraction of pigments from fresh Laminaria japonica

The pigments in Laminaria japonica was extracted with six organic solvents and analyzed in spectroscopy analysis. The extractions conditions were screened by an orthogonal test and the quantity of extracted pigments was determined spectroscopically. The results show that: (1) among the six organic solvents, acetone was the most effective one for the extraction; (2) the optimum extraction conditions were as follows: the ratio of S/M (solvent volume/ material weight) was 30 ml/g; fresh seaweed was extracted 2 times in 2 h; (3) the average total content of pigments was 1.85 mg/g (calculated with dry L. japonica).

Structural studies on a novel fucogalactan sulfate extracted from the brown seaweed Laminaria japonica

A low molecular weight fucogalactan, obtained from the brown seaweed Laminaria japonica, was separated into three fractions (LF1, LF2 and LF3) by DEAE-Sepharose FF column chromatography. All three fractions contained predominantly fucose, sulfate group and galactose. The results showed that the main fraction LF2 consisted of L-fucose, D-galactose and sulfate at a molar ratio 6:1:9. Structural study on the LF2 was carried out by NMR spectroscopy. The backbone of LF2 was primarily (1 -> 3)-linked alpha-L-fucopyranose residues (75%) and a few (1 -> 4)-alpha-L-fucopyranose linkages (25%). The branch points were at C-4 of 3-linked alpha-L-fucopyranose residues by beta-D-galactopyranose unites (35%, molar ratio) or at C-2 of 3-linked alpha-L-fucopyranose residues by non-reducing terminal fucose unites (65%, molar ratio). Sulfate groups occupied at position C-4 or C-2, sometimes C-2, 4 to fucose residues, and C-3 and/or C-4 to galactose residues. The structure of LF2 was supposed as following: [GRAPHICS] (C) 2010 Elsevier B.V. All rights reserved.

Optimal light regime for the cultivation of transgenic Laminaria japonica gametophytes in a bubble-column bioreactor

Fluctuating light intensity had a more significant impact on growth of gametophytes of transgenic Laminaria japonica in a 2500 ml bubble-column bioreactor than constant light intensity. A fluctuating light intensity between 10 and 110 mu E m(-2) s(-1), with a photoperiod of 14 h:10 h light:dark, was the best regime for growth giving 1430 mg biomass l(-1).

百合科黄精族的系统学研究

黄精族隶属于广义的百合科，通过对其研究历史的回顾，指出了它目前存在的问题，即(1)族的概念和范围不清、(2)属间的界限模糊、(3)族内各属的系统排列不明确．针对这些问题，本文从形态、解剖、胚胎、孢粉和细胞方面对族内各类群进行了野外调查和一些实验工作，本此基础上，并结合前人的工作，对黄精族的系统发育进行了探讨． 一, 形态本文通过黄精族各属大量标本，尤其是国产类群标本的认真研究，并结合6个多月的野外调查资料，分析了本族植物21个形态性状及其状态在属间和属内的分布，指出了一些性状的稳定性及其系统学价值， 二，解剖本文对黄精族5属17个种的茎、根状茎及子房进行了解剖学观察，同时对7属41种植物的叶表皮进行光镜观察，并对5属12种的叶表皮进行扫描电镜的观察，结果表明：(1)七筋姑属花葶的维管柱缺乏纤维鞘，这和本族其它类群不同．(2)扭柄花属、黄精属、竹根七属和鹿药属植物子房中具草酸盐针晶，而七筋姑属和万寿草属未发现这种晶体．(3)本族植物的气孔器均为无规则型，但邻近细胞及其周壁的形状在不同类群中是不同的，据此我们将本族叶表皮划分为七个类型，并认为叶表皮邻近细胞呈多边形、周壁直是黄精族叶表皮最基本的形式． 三，胚胎本文对黄精族3属4种植物以及铃兰(Convallaria majalis)进行了胚胎学研究，其中小玉竹(Polygonatum humile)和铃兰得到了完整的胚胎学资料，实验结果显示(1)黄精族植物小孢子发生过程中胞质分裂均为连续型，胚乳发育为核型;(2)小玉竹的胚囊发育方式为葱型，鹿药(Smilacina japonica)为英地百合型．万寿竹(Disporum cantoniense)和长蕊万寿竹(D. bodinicri)为蓼型，铃兰为葱型或英地百合型;(3)周缘细胞的有或无在小玉竹，鹿药和铃兰中是不稳定的． 四，孢粉本文对6属42种植物花粉进行了扫描电镜观察，结果表明(1)萌发孔的类型和数目在本族中不稳定，系统价值不大;(2)花粉外壁纹饰比较稳定，是一个十分重要的分类学性状．我们将黄精族植物的花粉外壁纹饰划分为9种类型．并认为孔穴型是一种原始类型，其它类型是由它演化而成， 五，细胞在前人工作的基础上，通过与其它性状的综合分析，本文认为黄精族植物有五种原始染色体基数：七筋姑属x=7;万寿竹属和扭柄花属x=8;黄精属x＝9：鹿药属和舞鹤草属x＝ 18;竹根七属x=20． 在上述实验结果的基础上，结合前人的工作，对黄精族的各种性状进行了分析，本文得出如下结论： 1．包括万寿竹属、扭柄花属、七筋姑属、黄精属、竹根七属、鹿药属和舞鹤草属的黄精族是一个单系类群． 2．本文赞同Dahlgren et al．(1985)，Tahktajan (1987)对黄精族系统位置的处理，即它与铃兰族，蜘蛛抱蛋族和沿阶草族关系较近，且共同组成铃兰科．相比之下，黄精族是该科中比较原始的一个族. 3．本文在性状分析的基础上，总结出20个比较稳定、系统价值较大的性状，并指出了它们的演化趋势;同时总结出黄精族原始类群应具有如下特征：(1).具细线形根状茎;(2).地上茎具分枝;(3).叶茎生;(4).具腋生的圆锥或总状花序;(5).花为三基数，花被片离生;(6).花被片具多条开放型脉;(7) .胚珠多数;(8).胚囊发育为单孢子型;(9).厚珠心胚珠;(10) .花粉外壁具覆盖层． 4．本文认为黄精族植物可分为三支：一是以染色体基数x＝9及其衍生的类群，包括黄精属，竹根七属、鹿药属和舞鹤草属;二是以x=8为基数的一支，它包括万寿竹属和扭柄花属;三是七筋姑属单独形成的一支，它的染色体基数x＝7．同时认为万寿竹属是本族比较原始的一个类群． 5．本文认为东亚北方是黄精族植物起源地． 6．本文赞同汤彦承(1978．见汪发缵、唐进1978)对卵叶扭柄花(Streptopus ovalis (Ohwi) Wang et Y.C．Tang)的分类学处理，而不赞同汪发缵、唐进(1983)把金佛山鹿药(Smilacina jinfoshanica Wang et Tang)从鹿药属转隶至黄精属的观点． 7．本文认为鹿药属和舞鹤草属亲缘关系较近，但区别特征明显，从而不赞同LaFrankie (1986),李恒(1990)将二者合并的观点。