Modulation of the Post-Prandial Plasma Glucose Profile in Juvenile European Sea Bass Dicentrarchus labrax Fed Diets Varying in Starch Complexity

The post-prandial glycemic response of juvenile European sea bass (initial mean weight 10 g) was determined following acclimation to experimental diets that contained a pea seed meal included at two levels. The ingredient provided a source of starch that replaced purified corn starcWyellow dextrin (2:1) in a reference diet based on fishmeal as the main protein component. The total carbohydrate levels amounted to 25% of the dry matter content and contributed (18%) to the overall gross dietary energy value of 20 MJ/kg. After a 12-wk feeding trial, in which the sea bass grew to over 40 g (mean weight), the plasma glucose profile was recorded over a 24-h period (0 h, 1 h, 3 h, 6 h, 9 h, 12 h, and 24 h) following a meal fed to satiation. Results showed that sea bass are able to regulate their glucostatic tolerance to dietary starch, and that the pea seed meal modulated this effect. Peak post-prandial responses varied between 1 to 9 h and were significantly delayed for the reference diet. Efficacy of glucose assimilation improved for the highest level of pea seed meal inclusion (40% by weight). The relevance of carbohydrate complexity with respect to feed formulation and nutritional requirements of sea bass are discussed.     

Structure and function of antimicrobial peptide penaeidin-5 from the black tiger shrimp Penaeus monodon

The gene for penaeidin-5, an antimicrobial peptide comprising 55 amino acids, was isolated from the hemocyte of black tiger shrimp (Penaeus monodon). RT-PCR expression tests revealed that penaeidin-5 was produced in hemocytes, gills, the intestine and muscle. Western blot analysis confirmed the panaeidin-5 was aboundantin hemocytes, the intestine and hemolymph. Immunohistochemistry revealedpenaeidin-5 in the cuticle and gills that are considered primary defense barriers. The deduced amino acid sequence of penaeidin-5 included a proline-rich N-terminal domain and a carboxyl-domain that contained six cysteine residues. Circular dichrosim analysis revealed an -helix in its secondary structure and the predicted 3D structure indicated two-disulfide bridges in the -helix. Based on the sequence of penaeidin-5 peptide cDNA, synthetic penaeidin-5 was prepared to carry out functional tests. The synthetic peptide had efficient bacteriostatic and bactericidal activity against Aerococcus viridans, and also inhibited the growth of two filamentous fungi, Fusarium pisi and Fusarium oxysporum. To measure penaeidin-5 in vivo, black tiger shrimp were challenged with Vibrio alginolyticus and A. viridans. At 3 h post-challenge, penaeidin-5 was induced and bacterial numbers decreased significantly by 12 h and 24 h.     

Comparison of early life history stages of the bay scallop, Argopecten irradians: Effects of microalgal diets on growth and biochemical composition

The culture of bay scallops, Argopecten irradians, is limited by a reliable and affordable supply of spat and the ability to ensure that animals attain market size within a single growing season. The main goals of our study were thus: (1) to develop growth-optimizing algal diets for implementation in hatcheries, and (2) to identify and compare bay scallop postlarval and juvenile dietary requirements, especially of lipids and fatty acids, which if met may enhance production. Nutritional needs of postlarval bay scallops (present study) are compared with those of sea scallops, Placopecten magellanicus, offered the same diets in a previous companion study. To this end, postlarval (initial shell height, SH = 240 μm) and juvenile (initial SH = 10 mm) bay scallops were offered 6–7 microalgal diet combinations at 20 °C, for 3 weeks. A similar growth ranking among diets was observed between the two developmental stages. A combination diet of Pavlova sp. (CCMP 459) and Chaetoceros muelleri was far superior to any other diet tested, yielding growth rates of 58 and 357 μm day^− 1 which were 65% and 25% higher than the next highest performing diet of Tetraselmis striata/C. muelleri in postlarvae and juveniles, respectively. The T. striata/C. muelleri diet, which is limited in the n-3 fatty acid docosahexaenoic acid (DHA), yielded very poor growth of sea scallop postlarvae in a prior study, indicating that bay scallops may have less stringent requirements for DHA than sea scallops. The Pav 459/C. muelleri diet, which also supported the highest growth of sea scallop postlarvae, is characterized by elevated levels of the n-6 fatty acids, arachidonic (AA) in C. muelleri and 4,7,10,13,16-docosapentaenoic (DPA) in Pav 459. The two diets deficient in AA and n-6 DPA, Pavlova lutheri/Thalassiosira weissflogii and P. lutheri/Fragilaria familica, yielded the lowest growth rates in both bay scallop postlarvae and juveniles. Tissue enrichment of these two fatty acids relative to the diet, as well as overall enrichment in ∑n-6 fatty acids was observed across developmental stages and dietary treatments. A similar pattern has previously been observed in sea scallop postlarvae, suggesting a dietary requirement for n-6 fatty acids in pectinids that has often been overlooked in the past.     

花鲈冷冻精液受精鱼苗与鲜精受精鱼苗的同工酶比较分析

采用水平凝胶电泳技术对花鲈(Lateolabrax maculatus)冷冻精液受精鱼苗与鲜精受精鱼苗进行了同工酶分析。本实验选取肌肉和肝脏两种组织,检测了IDHP、PGM、LDH、GPI、MDH、G3PDH、ME、HK、SOD、G6PD和SDH等11种酶,得到13个位点:IDHP*、PGM-1*、PGM-2*、LDH*、GPI-1*、GPI-2*、MDH*、G3PDH*、ME*、HK*、SOD*、G6PD*和SDH*。结果显示在0.99水平,SDH*和PGM-2*位点在两种受精鱼苗中均表现为多态,多态位点比例均为0.1538;平均预期杂合度分别为0.0460和0.0407,但未观测到杂合子,观测杂合度均为0,遗传偏离指数均为-1;有效等位基因数分别为1.0720和1.0850,花鲈冷冻精液受精鱼苗与鲜精受精鱼苗的遗传相似度为0.9991,遗传距离为0.0010。由此可见,花鲈的冷冻精液受精鱼苗和鲜精受精鱼苗间未出现同工酶水平上的遗传差异,冷冻保存精液完全适用于花鲈育苗生产。     

An improved enzyme preparation for rapid mass production of protoplasts as seed stock for aquaculture of macrophytic marine green algae

Preparation of protoplasts and their subsequent applications for both basic and applied research of marine macroalgae remains largely under developed due to lack of development of reliable methods with consistent yields of viable protoplasts. An improved enzyme preparation with a single commercial enzyme, e.g. 2% Cellulase Onozuka R-10 in 1% NaCl solution, was developed to produce protoplasts rapidly from different green algal genera of Ulva, Enteromorpha and Monostroma. The simple dissolution of enzyme powder in 1% NaCl resulted in exclusion of 2% Macerozyme R-10 from the mixture consisting of 2% Cellulase Onozuka R-10 with 3% NaCl earlier reported as superior for the same algae. Optimal conditions for the isolation of maximum yields of viable protoplasts were found to be with 2% Cellulase Onozuka R-10 incubated at 20 °C for 2 h in 1% NaCl solution with 0.8 M mannitol adjusted to pH 6.0. The protoplast yield with optimized enzyme mixture was as high as 102.8 × 10⁶ cells g^− 1 f. wt for M. oxyspermum while it was in the range of 74.4–88.6 × 10⁶ cells g^− 1 f. wt thallus for seven species of Ulva, and 82.5–95.4 × 10⁶ cells g^− 1 f. wt for three species of Enteromorpha. The regeneration rate of protoplasts isolated using this method ranged from 89 to 92% with normal morphogenesis. The seeding of nylon threads with isolated protoplasts of M. oxyspermum was successful and after 3–4 weeks the entire frame with nylon threads became thick green in color with tiny germlings in laboratory culture. Thus, the method described in the present study allow for rapid mass production of viable protoplasts that could be potentially used as a source for seed material for mariculture and for other applied phycological research.     

Furunculosis in Atlantic salmon (Salmo salar L.) is not readily controllable by bacteriophage therapy

The potential of bacteriophage therapy to control bacterial disease in farmed fish was tested using, as an example, furunculosis of Atlantic salmon, caused by Aeromonas salmonicida subsp. salmonicida.

In vivo testing with Atlantic salmon and rainbow trout (Oncorhynchus mykiss Walbaum) showed no adverse effects, with bacteriophage generally cleared within 96 h of administration by either intraperitoneal (i.p.) injection or oral in-feed.

Juvenile Atlantic salmon were administered a combination of bacteriophage O, R and B (1.9 × 10⁸ pfu fish^− 1) by i.p. injection, after they had been challenged with A. salmonicida subsp. salmonicida 78027, also by i.p. injection. The fish that were injected with bacteriophage immediately after challenge died at a significantly slower rate then those that were either not treated with bacteriophage, or treated 24 h post-challenge. However, the end result (100% mortality) was not affected.

In further experiments the effects of oral (1.88 × 10⁵ pfu g^− 1 fish^− 1 daily for 30 days), bath (1.04 × 10⁵ ml^− 1 daily for 30 days) and i.p. (6.25 × 10⁷ pfu fish^− 1) phage treatment to control furunculosis in experimentally infected Atlantic salmon were compared with antibiotherapy (treatment with 10 mg kg^− 1 bw^− 1 day^− 1 oxolinic acid for 10 days), using an indirect cohabitation challenge. No protection was offered by any of the bacteriophage treatments, compared to the positive challenge group, although significant protection was offered by the oxolinic acid treatment. Analysis of samples taken from the trials demonstrated that bacteriophage were correctly administered to the fish and, on occasion, were isolated from fish that had succumbed to furunculosis. It was also shown that bacteriophage resistant A. salmonicida subsp. salmonicida isolates could be recovered from mortalities in all the treatment groups.

The results suggest that, although there were no safety problems associated with the approach, furunculosis in Atlantic salmon is not readily controllable by application of bacteriophage.     

Dopamine modulates the physiological response of the tiger shrimp Penaeus monodon

Levels of glucose, lactate, pO₂, pCO₂, HCO₃⁻, TCO₂, Na⁺, K⁺, Cl⁻, protein, and oxyhemocyanin in the hemolymph and its osmolality and pH were measured when tiger shrimp, Penaeus monodon (13.5 ± 1.5 g body weight), were individually injected with saline or dopamine at 10^− 8, 10^− 7, or 10^− 6 mol shrimp^− 1. Results showed that hemolymph glucose, lactate, pCO₂, HCO₃⁻, and TCO₂ values increased from 2 to 4 h; hemolymph osmolality, Na⁺, and total protein had increased at 2 h; and hemolymph K⁺ decreased from 2 to 8 h after the dopamine injection. All physiological parameters returned to the control values 4–16 h after receiving dopamine. The dopamine injection also significantly decreased the oxyhemocyanin/protein ratio of P. monodon which occurred at 2 h, resulting from an elevation of hemolymph protein and a slight decrease of oxyhemocyanin. These results suggest that stress-inducing dopamine caused a transient period of modulation of energy metabolism, osmoregulation, respiration, and the acid–base balance in P. monodon in adapting to this environmental stress.     

Nutrients removed by Kappaphycus alvarezii (Rhodophyta, Solieriaceae) in integrated cultivation with fishes in re-circulating water

The potential of the red alga Kappaphycus alvarezii to remove nutrients was tested to treat effluents of Trachinotus carolinus fish cultivation, and the production of carrageenan in this condition was analyzed. Experiments were conducted in four tanks of 8000 L with approximately 1200 fishes of 30 g each integrated with three tanks of 100 L with 700 g of K. alvarezii, as initial biomass per tank. Seawater was re-circulated between tanks with seaweed and with fish. As a control, three tanks with seawater circulating in an open system were utilized. Seawater samples were collected daily for 10 days and concentrations of nitrate, nitrite, ammonium and phosphate were determined in the inflow and outflow water of the tanks. Significant differences between both collecting points were considered as nutrient removal by the seaweed. Growth rates and carrageenan yields were also analyzed in seaweed cultivated in seawater and in effluents. Growth rates of seaweed cultivated in tanks were lower than those obtained in open sea and in laboratory cultivation. Effluents had concentrations of nitrate and nitrite ca. 100 times higher than in the control. Maximum values of nutrient removal on effluents were: nitrate = 18.2%; nitrite = 50.8%; ammonium = 70.5% and phosphate = 26.8%. All plants survived throughout the experimental period, but some developed “ice–ice”, a disease associated with physiological stress. After the experimental period, some plants selected and cultivated in open sea presented higher growth rates in 40 days, indicating nutrient storage. No significant differences between carrageenan yields of K. alvarezii cultivated in seawater and in the effluents were observed. Our results show that K. alvarezii can be utilized as a biofilter for fish cultivation effluents, reducing the eutrophication process and can also be processed for carrageenan production, which provides an additional benefit to the fisheries.     

Evidences of the intertidal red alga Grateloupia turuturu in turning Vibrio parahaemolyticus into non-culturable state in the presence of light

Grateloupia turuturu, previously known as Grateloupia doryphora, has been widely reported to be an invasive algal species. There are no studies to relate the impact of its existence on its surrounding environment. In this paper, we present our results to show that about 70% of individuals collected from the field could turn Vibrio parahaemolyticus into non-culturable state on both selective (TCBS) and non-selective (2216E) culture medium in 24 h in the presence of light in live algal culture. Total bacteria counts on TCBS and 2216E plates dropped from the initial 565 (± 174) and 1192 (± 60) cfu ml^− 1 respectively to zero in 24 h. This effect disappeared when the alga was grown in darkness. The same effect was not found in two other intertidal macroalgae Laminaria japonica and Palmaria palmata. Further tests showed that the settlement ability of bacteria in seawater was impaired significantly in the presence of this alga in comparison with three other algal species.     

Pharmacokinetics and the active metabolite of enrofloxacin in Chinese mitten-handed crab (Eriocheir sinensis)

The pharmacokinetics and active metabolite of enrofloxacin were estimated after single intramuscular administration (10.0 or 20.0 mg/kg body weight) to the Chinese mitten-handed crab (Eriocheir sinensis) in fresh water at 25.0 ± 1.0 °C. Levels of enrofloxacin and its metabolite ciprofloxacin in the main tissues (hemolymph, hepatopancreas, muscle, ovary and spermary) were simultaneously detected by HPLC. Enrofloxacin concentration–time profiles for the hemolymph in both tests were described by a two-compartment open model with first-order absorption. Distribution half-time (T_1/2), elimination half-time (T_1/2β), body clearance (CL/F), mean residence time (MRT_0–∞), area under the concentration–time curve from 0 to ∞ h (AUC_0–∞) and apparent volume of distribution (Vd/F), which derived from the pharmacokinetic model, were 0.427 h, 21.3 h, 0.133 l/h/kg, 60.0 h, 96.9 μg/ml/h and 4.08 l/kg, respectively, at a dose of 10.0 mg/kg body weight, and 0.216 h, 12.3 h, 0.189 l/h/kg, 85.8 h, 187 μg/ml/h and 3.35 l/kg, respectively, at a dose of 20.0 mg/kg body weight. Similarities were found between the hemolymph concentration–time curves of the two tests; for example, instant absorption process followed by the distribution phrase, and a second absorption peak at 6 h post-treatment. After intramuscular administration of 10.0 mg/kg body weight, absorption of enrofloxacin was observed in the main edible tissues (hepatopancreas, muscle, ovary and spermary), and the drug residue was the highest in the hepatopancreas, where the ‘drug sink’ phenomenon occurred. Comparative pharmacokinetics showed fast absorption, broad distribution and fast elimination of enrofloxacin in E. sinensis after intramuscular dosing. Regarding ciprofloxacin, the main active metabolite of enrofloxacin, though relatively low levels were detected in all the main tissues of the crab, its kinetics in the hemolymph in the two tests were not described by a one- or two-compartment open model.     

Analysis of immune-relevant genes expressed in red sea bream (Chrysophrys major) spleen

Expressed sequence tag (EST) analysis is an efficient tool for gene discovery and for profiling gene expression. In order to isolate functional genes involved in immunity in fish, a cDNA library was constructed from red sea bream (Chrysophrys major) spleen by unidirectional cloning. A total of 2010 ESTs from the library was sequenced and compared with sequences in the GenBank database. Of the 2010 ESTs, 320 ESTs (15.9%) were identified as orthologs of known gene from other organisms by BLAST searches, whereas 1690 ESTs (84.1%) appeared to be unknown and are likely to represent newly described genes. These identified clones were derived from at least 81 genes, which were categorized into eight categories: 9 in cell structure/motility (11.1%), 14 in metabolism (17.3%), 8 in cell defense/immunity (10%), 5 in cell division (6.2%), 7 in cell signal transduction/communication (8.6%), 30 in gene/protein expression (37%), 5 hemoglobin (6.2%) and 3 genes lacking enough information to be classified (3.7%). Several important cDNAs involved in immune functions, such as immunoglobulin light chain (IgL), MHC class II, MHC class IIβ and RAP2c, were identified in red sea bream and compared for their structure with those from other organisms. Alignment showed that the red sea bream IgL precursor was closer to that of spotted wolfish than to that of yellowtail, Europe sea bass, orange spotted grouper, Atlantic salmon, channel catfish, fugu and sterlet. Phylogenetic analysis indicated that the red sea bream MHC II and MHC IIβ were more related to those from striped sea bass than to those from cichlid, flounder, salmonids, zebrafish and carp. High identity (over 92%) in deduced amino acid sequence of RAP2c between red sea bream and mammals implied that RAP2c gene was highly conserved during evolution.     

Effects of starvation and recovery on the survival, growth and RNA/DNA ratio in loliginid squid paralarvae

The ability of Loligo opalescens paralarvae to resist and recover from starvation was examined by measuring their survival, growth rate and RNA/DNA ratios during starvation and refeeding. Paralarvae were fed Artemia sp. nauplii, zooplankton and mysid shrimp. Fourteen days after hatching they were separated into five feeding treatments: a control treatment (food was always available) and treatments starved for 2, 3, 4 and 5 days, and then refed. Each day, 5–7 paralarvae from each treatment were anesthetized to measure mantle length and wet weight (WW), and then RNA and DNA were extracted using an ethidium bromide fluorometric technique. Paralarvae did not survive 4 and 5 days of starvation, showing that at 15 days of age and at 16 °C the limit to recovery was 3 days of starvation. Paralarvae starved for 2 and 3 days showed compensatory growth that mitigated the effects of starvation, in that at the end of the experiment (10 days), they attained mean final body weights similar to the control treatment. Differences in the RNA/DNA ratios between control and starved paralarvae were detected within 2 days of food deprivation. For paralarvae starved 2 and 3 days, it took 1 day after refeeding to attain RNA/DNA ratios not significantly different from the control treatment. Additionally, RNA/DNA ratios were highest during the day (0800, 1200, 1600 h) and lowest at night (0000, 0400 h), suggesting daytime feeding activity. Growth rates ranged from − 14% to 21% WW day^− 1 and the resulting equation between RNA/DNA ratio and growth rate (GR) of paralarvae was GR = 1.74 RNA/DNA − 11.79 (R² = 0.70). After starvation, there was a reduction in growth variability in all starved treatments, while growth variability remained high in the control treatment. Findings from the present study indicate that nucleic acids are a valid indicator of nutritional condition and growth in squid paralarvae.     

Diurnal variation of tryptic activity in larval stage and development of proteolytic enzyme activities of malabar grouper (Epinephelus malabaricus) after hatching

Diurnal variation in tryptic activity and developmental changes in proteolytic enzyme activities of malabar grouper larvae (Epinephelus malabaricus) were examined. Five different groups were prepared for the experiment of diurnal variation of tryptic activity in larvae: larvae were fed Thai-type rotifers Brachionus rotundiformis from the time of mouth opening, fed rotifers from 6 h after mouth opening, 12 h, 24 h and not fed rotifers (starved control). The experimental tanks were placed in temperature-controlled baths at 28 °C under 24 h light. Developmental changes in proteolytic activity of trypsin and pepsin-like enzyme were measured from hatching to 57 days after hatching (DAH).

The tryptic activity of all fed groups showed the same pattern, and the diurnal variation of tryptic activity was clearly observed from 3 to 6 DAH. The highest tryptic activities were found at 19:00, and the activities were lowest from 01:00 to 07:00. In contrast, that of non-fed larvae was low compared to the fed groups, however the diurnal variation of tryptic activity was shown same tendency to the fed groups. Interestingly, both groups (fed and non-fed) were exhibited a circadian rhythm under the 24 h light conditions and delaying of first-feeding. Tryptic activity of larvae notably increased from 40 to 45 DAH and markedly decreased at 52 DAH. In contrast to the tryptic activity, that of pepsin-like enzyme clearly increased from 47 to 51 DAH. The results suggest that a functional change of protein digestion occurs from 40 to 50 DAH related with metamorphosis in malabar grouper. These results could contribute to determining appropriate feeding schedules, such as feeding time, frequency and optimal time to change food items, in mass-scale production of the present species.     

Melanocyte-stimulating hormone facilitates hypermelanosis on the non-eyed side of the barfin flounder, a pleuronectiform fish

A pleuronectiform fish, the barfin flounder, Verasper moseri, is promising for aquaculture and resource enhancement in Northern Japan due to its high commercial value. Hypermelanosis of its non-eyed side, which frequently occurs under culture conditions, diminishes its commercial value. Two peptide hormones, melanin-concentrating hormone (MCH) and melanocyte-stimulating hormone (MSH), having opposing actions, are associated with the color changes of fish. We have previously reported the positive effect of MCH in preventing hypermelanosis. Here, we examined the effects of MSH on the occurrence of hypermelanosis. A single injection of Des-Ac--MSH [0.01 nmol/g–10 nmol/g (0.016 μg–16 μg/g)] did not change the eyed-side body color, while a single injection of MCH [0.1 nmol/g (0.21 μg/g)] made the eyed-side color paler. No difference was observed in eyed-side lightness between fish injected with MCH (0.1 nmol/g) and those receiving MCH (0.1 nmol/g) and an increased amount of Des-Ac--MSH (0.01 nmol/g–10 nmol/g) simultaneously. These results indicate that MSH does not suppress the in vivo body color-paling effects of MCH in barfin flounders. On the other hand, implantation of a cholesterol pellet containing Des-Ac--MSH (280 μg, twice at 29-day interval) increased hypermelanosis of the non-eyed side of barfin flounders compared to control fish. Eyed-side bodies of MSH-treated fish were darker than control fish; thus, MSH is involved in morphological color change including ectopic melanin synthesis in non-eyed-side skin.     

Effect of increasing salinity on physiological response in juvenile scallops Argopecten purpuratus at two rearing temperatures

Argopecten purpuratus can be cultivated using Recirculating Aquaculture Systems (RAS) as a method to increase production. In order to determine physiological response of A. purpuratus under different salinities and temperature conditions, two groups of juvenile scallops (small: h = 6.5 mm and large: h = 25.5 mm) were acclimated and close-cultured at salinities of 34, 38, and 42 g/l, at 16 and 22 °C and fed on Isochrysis galbana and Chaetoceros calcitrans. Survival, shell growth and scope for growth were determined at the end of the trials. Survival showed an inverse relationship with temperature and ammonia levels. In small scallops an increase in salinity at 16 °C increased survival. However, this relationship was not evident at 22 °C. On the other hand, salinity did not affect survival of large juveniles. Small juveniles had a lower survival (approximately 40%) than larger scallops (up to 85%) throughout the trials. Oxygen consumption was not affected by salinity. Small scallops showed similar oxygen consumption at 16 and 22 °C but in large juveniles higher values were registered at 22 °C. In large juveniles routine consumption at 16 °C was higher (up to 35%) than standard consumption. This pattern was not evident at 22 °C, suggesting that oxygen demand is higher regardless of feeding condition. NH₄⁺–N excretion rate is inversely related to salinity. Only small juveniles showed a higher NH₄⁺–N excretion rate at 22 °C. Scope for growth was positive in all treatments, although the upper limit of salinity should not be based only in this index. Higher scope for growth values at 38 and 42 g/l was related with a reduction in ammonia excretion and high absorption efficiency. In addition, an increase in salinity produced a reduction in NH₃–N proportion and under hypersaline conditions scallops tended to decrease excretion as a way of osmoconformation. This explains our findings of higher survival rates at higher salinities. Even though the scope for growth is positive at 42 g/l, the osmotic stress reduces the survival chances. The data obtained can be considered useful information for A. purpuratus culture under controlled conditions.     

海鲈游泳续航时间和运动代谢规律研究

为研究海鲈(Lateolabrax japonicus)的游泳续航能力和游泳过程中的物质代谢规律,以深水网箱养殖的3月龄海鲈为研究对象,采用游泳续航时间和生化指标作为评测海鲈游泳能力和代谢规律的方法进行了试验。结果显示,海鲈的游泳续航时长为200 min,其续航能力强于大黄鱼而弱于美国红鱼。在游泳过程中,海鲈血糖浓度升高,但差异性不显著(P>0.05),肌糖原随着时间的增加显著减少(P<0.05),且肌糖原浓度在海鲈疲劳时接近于0。当鱼接近100%疲劳时,其肝糖原浓度也接近于0;当海鲈以不同疲劳程度游泳时,其肌肉乳酸脱氢酶浓度会显著升高(P<0.05)。表明海鲈的续航游泳过程伴随着无氧呼吸和有氧呼吸。     

Effect of low water temperature on viral replication of white spot syndrome virus in Procambarus clarkii

This study evaluated the effect of low water temperature (10 ± 1 °C) on viral infection and replication of white spot syndrome virus (WSSV) in crayfish, Procambarus clarkii, under standardized conditions. Crayfish were (i) maintained at 24 ± 1 °C before challenge and 10 ± 1 °C afterwards, or (ii) maintained at 10 ± 1 °C before challenge and 24 ± 1 °C afterwards. No mortality was observed when crayfish were held at 10 ± 1 °C after challenge, but mortality reached 100% when they were transferred to 24 ± 1 °C. Competitive PCR showed that viral levels at 10 ± 1 °C rose from 10⁶ to 10⁸ copies/mg of gill tissues, while at 24 ± 1 °C levels increased from 10⁶ to 10¹⁰ copies/mg of gill tissues during the same time interval. These results showed that a low water temperature of 10 ± 1 °C could reduce viral replication when compared to 24 ± 1 °C but could not prevent it.     

Purification and partial characterization of Cu, Zn superoxide dismutase from haemolymph of Oriental river prawn Macrobrachium nipponense

The copper plus zinc superoxide dismutase (Cu, Zn-SOD) was purified from haemolymph of the Oriental river prawn, Macrobrachium nipponense and partially characterized. Partial protein precipitation in crude extract was affected by using heat treatment and (NH₄)₂SO₄ fractionated precipitation methods. Fractionation of superoxide dismutase was performed by DEAE-cellulose 32 ion-exchange chromatography and followed by CM-cellulose cation-exchange chromatography. The molecular weight of it was about 66.1 kDa, as judged by SDS polyacrylamide gel electrophoresis. The enzyme was sensitive to cyanide and H₂O₂, and contained 1.08 ± 0.14 atom of copper and 0.98 ± 0.11 zinc per subunit shown in atomic absorption spectroscopy, which revealed that purified SOD was Cu, Zn superoxide dismutase. The purified enzyme had an absorption peak of 269 nm in ultraviolet region and the enzyme remained stable at 25–45 °C within 60 min. But it was rapidly inactivated at higher temperature (50 °C). The activity of purified shrimp Cu, Zn-SOD was remained stable over the range pH 5.8–8.3. Treated with 10 mM mercaptoethanol, the enzyme activity significantly increased. However, the enzyme activity was obviously inhibited by 10 mM CaCl₂, ZnCl₂, SDS, EDTA–Na₂ and 1 mM and 10 mM K₂Cr₂O₇. The results showed that it might be a kind of EC-SOD. And it was the first report of some characterizations of this EC-SOD in M. nipponense.     

Effect of tremata closures on the oxygen consumption rhythm of ezo abalone Haliotis discus hannai

Cultured adults of ezo abalone, Haliotis discus hannai (shell length, 89.3 ± 6.4 mm and flesh dry weight, 13.0 ± 1.6 g; n = 12) were exposed to different temperatures (10 and 25 °C) to determine whether respiration effects were induced by artificial closure of the second and third tremata (respiratory pores). A respirometer was used to determine the oxygen consumption rates (OCRs) as measures of metabolic activity. The closed tremata were the second and third of the four open tremata anterior to the head of the abalone. The OCRs of starved abalone were measured under constant conditions (CC: constant dark and constant temperature) during a 240-h period, consisting of 120 h before and 120 h after the closure of tremata. The endogenous rhythm of the OCRs in cultured ezo abalone exhibited a dominant circadian rhythm (unimodal rhythm) in the latter half of the experimental period and occasionally showed a weaker but similar circatidal rhythm (bimodal rhythm) in the first half of the experimental period regardless of temperature. The results from the present study indicate that the rhythmicity of the OCRs in starved abalone is not affected by closure of the second and third tremata. This study offers essential physiological information for utilizing tremata in developing a tagging technique in abalone.     

Optimal conditions for egg storage, incubation and post-hatching growth for the freshwater turtle, Chelodina rugosa: Science in support of an indigenous enterprise

Incubation of northern snake-necked turtle (Chelodina rugosa) eggs and subsequent sale of hatchlings for the pet industry has the potential to provide culturally suitable employment for indigenous communities in northern Australia. Developmental arrest in response to egg inundation is unique to C. rugosa. Eggs can be stored under water for up to 10 weeks without appreciable impact on egg or embryo survival, allowing the transport and sale of eggs into niche markets without high levels of mortality, and permitting eggs to accumulate in diapause until there are sufficient numbers to incubate as batches. Eggs that are not inundated or inundated for short periods experience similar survival rates to eggs inundated for lengthy periods. Incubation temperature influences embryo survival and development period in C. rugosa. Embryonic survival is greatest at 26 °C, steadily declining as temperature increases to 32 °C. A similar increase in incubation temperature decreases incubation period by approximately 40 days, however almost half of this variation is attributed to the increase in incubation temperature from 26 to 28 °C. Hatchling growth in C. rugosa is characterized by two phases. There is an initial phase of relatively slow growth under the partial influence of initial egg size and incubation duration, followed by a second phase of relatively rapid growth under the partial influence of water temperature and mass at hatching. Post-hatching survival is negatively correlated with duration of egg inundation and water temperature. Evidence suggests that inundation of C. rugosa eggs for 6 weeks, incubation of embryos at 28 °C and raising hatchlings in 28 °C water will yield the best overall outcomes.