Application of Lactic Acid Bacteria (LAB) in Freshness Keeping of Tilapia Fillets as Sashimi

Aquatic products are extremely perishable food commodities. Developing methods to keep the freshness of fish represents a major task of the fishery processing industry. Application of Lactic Acid Bacteria(LAB) as food preservative is a novel approach. In the present study, the possibility of using lactic acid bacteria in freshness keeping of tilapia fillets as sashimi was examined. Fish fillets were dipped in Lactobacillus plantarum 1.19(obtained from China General Microbiological Culture Collection Center) suspension as LAB-treated group. Changes in K-value, APC, sensory properties and microbial flora were analyzed. Results showed that LAB treatment slowed the increase of K-value and APC in the earlier storage, and caused a smooth decrease in sensory score. Gram-negative bacteria dominated during refrigerated storage, with Pseudomonas and Aeromonas being relatively abundant. Lactobacillus plantarum 1.19 had no obvious inhibitory effect against these Gram-negatives. However, Lactobacillus plantarum 1.19 changed the composition of Gram-positive bacteria. No Micrococcus were detected and the proportion of Staphylococcus decreased in the spoiled LAB-treated samples. The period that tilapia fillets could be used as sashimi material extended from 24 h to 48 h after LAB treatment. The potential of using LAB in sashimi processing was confirmed.     

Development of an inactivated iridovirus vaccine against turbot viral reddish body syndrome

Turbot(Scophthalmus maximus L.) reddish body iridovirus(TRBIV) was propagated in turbot fin cells(TF cells) and inactivated as the TRBIV vaccine with its protection efficiency evaluated in this study.TF cells were cultured in 10% bovine calf serum(BCS)-containing MEM medium(pH7.0) at 22℃,in which TRBIV propagated to a titer as high as 105.6 TCID50 mL-1.The TRBIV was inactivated with 0.1% formalin and formulated with 0.5% aluminum hydroxide.The inactivated vaccine caused neither cytopathogenic effect(CPE) on TF cells nor pathogenic effect on turbots.After being administered with the vaccine twice via muscle injection,the turbot developed high-tittered TRBIV neutralizing antibodies in a dose-dependent manner.The vaccine protected the turbot from dying with an immunoprotection rate of 83.3% as was determined via subcutaneous vaccination in the laboratory and 90.5% via bath vaccination in turbot farms,respectively.The inactivated vaccine was very immunogenic,efficiently preventing tur-bot from death.It holds the potential of being applied in aquaculture.     

Distribution and ultrastructure of pigment cells in the skins of normal and albino adult turbot, Scophthalmus Maximus

The distribution and ultrastructure of pigment cells in skins of normal and albino adult turbots were examined with transmission electron microscopy (TEM). Three types of pigment cells of melanophore, iridophore and xanthophore have been recognized in adult turbot skins. The skin color depends mainly on the amount and distribution of melanophore and iridophore, as xanthophore is quite rare. No pigment cells can be found in the epidermis of the skins. In the pigmented ocular skin of the turbot, melanophore and iridophore are usually co-localized in the dermis. This is quite different from the distribution in larvae skin. In albino and white blind skins of adult turbots, however, only iridophore monolayer still exists, while the melanophore monolayer disappears. This cytological evidence explains why the albino adult turbot, unlike its larvae, could never resume its body color no matter what environmental and nutritional conditions were provided. Endocytosis is quite active in the cellular membrane of the iridophore. This might be related to the formation of reflective platelet and stability of the iridophore.     

Kinetic study of alkaline protease 894 for the hydrolysis of the pearl oyster Pinctada martensii

A new enzyme (alkaline protease 894) obtained from the marine extremophile Flavobacterium yellowsea (YS-80-122) has exhibited strong substrate-binding and catalytic activity, even at low temperature, but the characteristics of the hydrolysis with this enzyme are still unclear. The pearl oyster Pinctada martensii was used in this study as the raw material to illustrate the kinetic properties of protease 894. After investigating the intrinsic relationship between the degree of hydrolysis and several factors, including initial reaction pH, temperature, substrate concentration, enzyme concentration, and hydrolysis time, the kinetics model was established. This study showed that the optimal conditions for the enzymatic hydrolysis were an initial reaction pH of 5.0, temperature of 30°C, substrate concentration of 10% (w/v), enzyme concentration of 2 500 U/g, and hydrolysis time of 160 min. The kinetic characteristics of the protease for the hydrolysis of P. martensii were obtained. The inactivation constant was found to be 15.16/min, and the average relative error between the derived kinetics model and the actual measurement was only 3.04%, which indicated a high degree of fitness. Therefore, this study provides a basis for the investigation of the concrete kinetic characteristics of the new protease, which has potential applications in the food industry.     

Effect of stocking density on performances of juvenile turbot (Scophthalmus maximus) in recirculating aquaculture systems

Limited information has been available about the influence of loading density on the performances of Scophthalmus maximus, especially in recirculating aquaculture systems (RAS). In this study, turbot (13.84±2.74 g; average weight±SD) were reared at four different initial densities (low 0.66, medium 1.26, sub-high 2.56, high 4.00 kg/m²) for 10 weeks in RAS at 23±1°C. Final densities were 4.67, 7.25, 14.16, and 17.47 kg/m², respectively, which translate to 82, 108, 214, and 282 percent coverage of the tank bottom. Density had both negative and independent impacts on growth. The final mean weight, specific growth rate (SGR), and voluntary feed intake significantly decreased and the coefficient of variation (CV) of final body weight increased with increase in stocking density. The medium and sub-high density groups did not differ significantly in SGR, mean weight, CV, food conversion rate (FCR), feed intake, blood parameters, and digestive enzymes. The protease activities of the digestive tract at pH 7, 8.5, 9, and 10 were significantly higher for the highest density group, but tended to be lower (not significantly) at pH 4 and 8.5 for the lowest density group. The intensity of protease activity was inversely related to feed intake at the different densities. Catalase activity was higher (but not significantly) at the highest density, perhaps because high density started to induce an oxidative effect in turbot. In conclusion, turbot can be cultured in RAS at a density of less than 17.47 kg/m². With good water quality and no feed limitation, initial density between 1.26 and 2.56 kg/m² (final: 7.25 and 14.16 kg/m²) would not negatively affect the turbot cultured in RAS. For culture at higher density, multi-level feeding devices are suggested to ease feeding competition.     

Pharmacokinetics and acetylation of sulfamethoxazole in turbot <Emphasis Type="Italic">Scophthalmus maximus</Emphasis> after intravascular administration

The pharmacokinetic profiles and sulfamethoxazole (SMX) acetylation process in turbot reared at 18°C were investigated. Either SMX (parent drug) or its acetylized metabolite, N₄-acetylsulfamethoxazole (AcSMX), was administered intravascularly to turbot at a dosage of 50 mg/kg BW. Serum concentrations of the parent drug and its metabolite were both measured by HPLC, and the changes in concentration over time were analyzed in two- and non-compartment models because SMX treatment produced multiple peaks. The results demonstrated that the elimination half-life of the parent drugs, SMX and AcSMX, were 159.2 and 5.9 h, respectively. The apparent volume of distribution was 0.2 and 0.8 L/kg, and the clearance was 0.038 and 0.222 L/(h·kg), for SMX and AcSMX, respectively. SMX acetylation in turbot was 2.8%, and the deacetylation of AcSMX was 0.2%. These findings may be useful in optimizing SMX dosage regimens in turbot aquaculture.     

Comparison of the morphometric dynamics of fast-growing and slow-growing strains of turbot Scophthalmus maximus

The dynamics of changes in body shape of fast-growing and slow-growing strains of turbot Scophthalmus maximus, and of the differences in body shape between the two strains, were evaluated from 3 to 27 months of age. The ratios of total length/body length, body width/body length and total length/body width were used as morphometric indices. The two strains exhibited different temporal trends in total length/body length but similar trends in body width/body length and total length/body width. Generally, body width/body length of the two strains increased with time and total length/body width decreased. Thus, the bodies of both fast-growing and slow-growing strains of turbot changed from a narrow to a more rounded shape. However, the ratio total length/body length was generally lower, body width/body length was mostly higher and total length/body width was consistently lower in the fast-growing strain than in the slowgrowing strain. Correlation analysis of the three shape ratios with body weight showed that total length/body length and total length/body width were unsuitable, and that width/body length was suitable, for use as a phenotypic marker for selective breeding of turbot for growth in weight.     

Impact assessment of modified clay on embryo-larval stages of turbot Scophthalmus maximus L.

Using modified clay is one of the most promising methods for the mitigation of harmful algal blooms(HABs). However, the environmental impact of modified clay has become a subject of global concern. In this study, turbot(Scophthalmus maximus L.) embryos were used as a model to assess the effect of modified clay on this sensitive stage of fish development. Results show that the 24 and 48 h LC_(50)(median lethal concentrations) of the modified clay were 1.70 and 1.65 g/L, and the safe concentration was 0.47 g/L, which is much higher than the concentration used to treat HAB. The modified clay did not affect significantly the hatchability of turbot embryos but when the concentration exceeded 0.50 g/L, the deformity rate of newly hatched larvae increased significantly. The total length, specific growth rate(SGR)and yolk sac absorption rate of larvae reached their peaks at 0.50 g/L and then gradually decreased as the concentration of modified clay increased. Therefore, a moderate amount of modified clay does not harm the survival and hatching of turbot embryos, or the growth and development of newly hatched larvae.     

Effects of waterborne Fe(II) on juvenile turbot Scophthalmus maximus: analysis of respiratory rate, hematology and gill histology

The concentration of Fe(Ⅱ) is high in some groundwater supplies used in turbot culture,and the toxicity of waterborne Fe(Ⅱ) is unknown.We investigated the stress responses of juvenile turbot,Scophthalmus maximus,exposed to Fe(Ⅱ) of different concentrations (0.01,0.05,0.1,0.5,1,and 2 mg/L) for 1,7,14,and 28 d,under the same ambient conditions of other parameters.Changes in respiratory rate,hematological parameters,and gill structure were determined.The results show that waterborne Fe(Ⅱ) did not cause severe hematological perturbation to turbot.A low-medium Fe(Ⅱ) concentration (lower than 0.1 mg/L) could boost the respiratory rate,and caused no or very limited damage to fish.A high Fe(Ⅱ) concentration (0.1 mg/L or higher),however,caused gill damage,such as vacuoles in branchial lamellae,epithelial necrosis,and hypertrophy of epithelial cells,and even death after extended exposure time.Therefore,excess waterborne Fe(Ⅱ) and long-term exposure to Fe(Ⅱ) could be responsible for poor growth and high mortality of turbot in culture.The concentration of waterborne Fe(Ⅱ) in turbot culture should be kept below 0.1 mg/L.     

Molecular characterization and expression analysis of Lily-type lectin (SmLTL) in turbot Scophthalmus maximus,and its response to Vibrio anguillarum

A full-length lily-type lectin(Sm LTL) was identified from turbot(Scophthalmus maximus) in this study. By searching database for protein identification and function prediction, Sm LTL were confirmed. The full-length c DNA of Sm LTL is composed of 569 bp and contains a 339 bp ORF that encodes 112 amino acid residues. The Sm LTL peptide is characterized by a specific β-prism architecture and contains three mannose binding sites in a three-fold internal repeat between amino acids 30–99; two of the repeats share the classical mannose binding domain(Qx Dx Nx Vx Y) while the third binding site was similar to other fish-specific binding motifs(Tx Tx Gx Rx V). The primary, secondary, and tertiary structures of Sm LTL were predicted and analyzed, indicating that the S m LTL protein was hydrophilic, contained 5.36% α-helices, 39.29% extended strands, 16.07% β-folds, and 39.29% random coils, and three β-folds. Quantitative realtime polymerase chain reaction(qPCR) analysis revealed that the Sm LTL mRNA was abundantly expressed in skin, gill, and intestine. Low levels of Sm LTL expression were observed in other tissues. The expression of Sm LTL in gill, skin and intestine increased at m RNA level after stimulation of V ibrio anguillarum, our results suggest that Sm LTL serve as the first line of defence against microbial infections and play a pivotal role in the innate mucosal immune system. The current study indicates that Sm LTL is a member of the lilytype lectin family and the information reported here will provide an important foundation for future research on the role of this protein.     

Ciliate Uronema marinum is the causative agent of scuticociliatosis in farm raised turbot Scophthalmus maximus

To identify the pathogen that causes scuticociliatosis in farmed-raised turbot Scophthalmus maximus,we isolated a ciliate from the brain tissue of an infected turbot and identified it as Uronema marinum based on morphological and molecular evidence.We then infected the turbots in artificial laboratory settings with pure cultured U.marinum.The infected turbots showed syndromes similar to those observed in naturally infected ones.Furthermore,microscopic examination and PCR detection confirmed the presence of the ciliate in the tissues of those laboratory-infected turbots.To our best knowledge,this is the first report of scuticociliatosis caused by U.marinum in farm-raised turbot S.maximus in China.     

开鲁盆地陆西凹陷热埋藏史研究

本文通过对有机质干酪根类型的分析，重新确定了陆面凹陷时间温度指数（ＴＴＩ）值计算的基准温度及古地温梯度，对凹陷内沉积厚度大的地方进行了沉积构造史的恢复，并建立了凹陷热埋藏史模式．据此计算了凹陷内部分ＴＴＩ值．通过研究，指出了目前对凹陷进行石油勘探的有利生油岩层位．     

Cloning and sequence analysis of a full-length cDNA of SmPP1cb encoding turbot protein phosphatase 1 beta catalytic subunit

Reversible protein phosphorylation, catalyzed by protein kinases and phosphatases, is an important and versatile mechanism by which eukaryotic cells regulate almost all the signaling processes. Protein phosphatase 1 (PP1) is the first and well-characterized member of the protein serine/threonine phosphatase family. In the present study, a full-length cDNA encoding the beta isoform of the catalytic subunit of protein phosphatase 1(PP1cb), was for the first time isolated and sequenced from the skin tissue of flatfish turbot Scophthalmus maximus, designated SmPP1cb, by the rapid amplification of cDNA ends (RACE) technique. The cDNA sequence of SmPP1cb we obtained contains a 984 bp open reading frame (ORF), flanked by a complete 39 bp 5' untranslated region and 462 bp 3' untranslated region. The ORF encodes a putative 327 amino acid protein, and the N-terminal section of this protein is highly acidic, Met-Ala-Glu-Gly-Glu-Leu-Asp-Val-Asp, a common feature for PP1 catalytic subunit but absent in protein phosphatase 2B (PP2B). And its calculated molecular mass is 37 193 Da and pI 5.8. Sequence analysis indicated that, SmPP1cb is extremely conserved in both amino acid and nucleotide acid levels compared with the PP1cb of other vertebrates and invertebrates, and its Kozak motif contained in the 5'UTR around ATG start codon is GXXAXXGXXATGG, which is different from mammalian in two positions A-6 and G-3, indicating the possibility of different initiation of translation in turbot, and also the 3'UTR of SmPP1cb is highly diverse in the sequence similarity and length compared with other animals, especially zebrafish. The cloning and sequencing of SmPP1cb gene lays a good foundation for the future work on the biological functions of PP1 in the flatfish turbot.     

Pharmacokinetics and tissue behavior of enrofloxacin and its metabolite ciprofloxacin in turbot Scophthalmus maximus at two water temperatures

Turbot Scophthalmus maximus, an important aquaculture species in China, currently suffers from epizootic diseases because of high density aquaculture. Enrofloxacin has been used to treat various systemic bacterial fish infections. However, studies concerning the pharmacokinetics of enrofloxacin in turbot are limited. In this study, the pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin, were investigated in the turbot following intravenous and oral administration at 10 mg enrofloxacin/kg body weight, at 16°C and 10°C water temperatures. The concentrations of enrofloxacin and ciprofloxacin in the main tissues (plasma, muscle, liver and kidney) were detected by HPLC. The results show that the plasma concentration-time data for enrofloxacin were best described as a two-compartment open model after intravenous and oral administration. Three pharmacokinetic equations were established between the concentrations and temperatures. The kinetic profile of enrofloxacin was temperature dependent. The absorption half-life of enrofloxacin was 1.99 h and 2.17 h after oral administration, whereas the elimination half-life of the drug was 98.63 h and 136.59 h at 16°C and 10°C, respectively. The peak concentration of enrofloxacin in plasma and tissues was higher at 16°C than that at 10°C, and the peak plasma concentration time in the liver was the shortest at both temperatures among those of other tissues. The plasma C max /MIC ratio varied between 11.08 and 5 540.00 at 16°C; and between 7.92 and 3 960.00 at 10°C. The AUC/MIC ratio was 467.82-280 690.00 at 16°C, and 359.48-215 690.00 at 10°C. These ratios indicate that it is possible to obtain therapeutic efficacy. Very low levels of ciprofloxacin were detected. The AUC ratios of ciprofloxacin and enrofloxacin in plasma suggest that plasma ciprofloxacin might play a minor role in enrofloxacin treatment for turbot.     

基于底拖网调查数据的渔业资源质量状况评价体系构建——以北部湾为例

【目的】构建渔业资源质量状况评价体系,评估渔业管理成效,服务海洋渔业可持续发展。【方法】以北部湾底拖网调查数据为基础,结合渔业统计数据,构建渔业资源质量状况评价指标体系。该体系分3个层次,下设7项指标。通过专家经验法和平均赋值法对指标进行赋值,得到各指标权重,建立质量状况评价等级指数,并利用该等级指数对北部湾渔业资源进行评价。【结果与结论】1992―2017年北部湾渔业资源评价指标中有6项呈下降趋势;质量状况评价等级指数波动在0.44~0.56之间。自20世纪90年代以来,北部湾渔业资源一直处于临界状态。该评价结果与北部湾渔业资源利用的实际状况基本相符,证明该方法具有合理性和可操作性,可为其他海区渔业资源质量状况评价提供借鉴。     

Isolation,Identification, and Quantitative Determination of Saponin in Apostichopus japonicus by HPLC-DAD

Apostichopus japonicus is an important invertebrate that is widely used as a tonic food in Asian countries.The purpose of this study is to purify and identify a class of compound,the saponins,from the body wall of A.japonicus,and to establish a new me-thod to determine the quantity of saponins in the sea cucumber.In this study,the saponins of A.japonicus,cladoloside A(CA),were ob-tained from 80%ethanol extract by column chromatography for the first time and were characterized using the spectral method.The resulting purified saponins were then profiled using ¹HNMR,¹³CNMR,and ESI-MS,which revealed the CA molecular formula of C₅₃H₈₂O₂ and contained a triterpenoid backbone,a methylglucopyranosyl moiety,a quinovopyranosyl,and two xylopyranosyls.A me-thod for the quantitative determination of CA,comprising microwave-assisted extraction,high-performance liquid chromatography,and diode array detector method,was established.Extraction efficiency was optimized by changing microwave power,extraction sol-vent,volume,time,and temperature.Results showed that under the optimum conditions(extraction time of 10 min,temperature of 45℃,and solvent of 25 mL 70%ethanol under 400 W),the detection limit of CA was 0.0015 mg mL^?1,and the recoveries of CA from samples at spiking levels of 10,20,and 50μg g^?1 ranged from 90.1%-104%.The proposed method was successfully applied to ana-lyze the saponins in different tissues of A.japonicus collected in different seasons.The method developed in this study can provide quantitative technical support for the quality control of A.japonicus.     

The eco-nutrition requirements for dietary protein and its rhomb characteristics in juvenile turbot (Scophthalmus maximus L.)

We evaluated the dietary protein requirements of juvenile turbot (Scophthalmus maximus L.) and their effects on aquatic quality. Five experimental diets were formulated containing 450, 480, 500, 520, and 540 g/kg. Each diet was randomly assigned to triplicate groups of juvenile turbot (mean initial body weight 34.5 ± 5.5 g) for 88 d. Both the weight gain ratio and feed efficiency increased with increasing dietary protein up to 500 g/kg, but no further improvement was detected when dietary protein levels were >500 g/kg. Protein intake and digestion increased with protein levels, while fecal nitrogen and nitrogen content in seawater increased only when dietary protein exceeded 500 g/kg. Protein digestibility was highest at intermediate dietary protein levels. Chemical oxygen demand, nitrite-nitrogen (NO2--N) and phosphatic-phosphor (PO43--P) levels increased in the rearing water as dietary protein levels increased. The optimum eco-nutrition level of dietary protein for juvenile turbot was 500 g/kg under the current experimental conditions. The diets containing 540 and 500 g/kg protein had similar growth rates and feed conversion ratios, but levels of ammonia (NH4+) and nitrogen were considerably higher in the water and feces, respectively, at the higher level of dietary protein. The difference in the pattern of change between body weight gain and ammonia concentration in water with increasing dietary protein is described by rhomb characteristics.     

神经网络法反演海水叶绿素浓度的分析

海洋水色遥感的最终目的之一是监测海洋初级生产力的时空变化,而反映海洋初级生产力的一个重要指标就是浮游植物中的叶绿素浓度[1]。对于海水叶绿素浓度的遥感反演研究工作至今已进行了30多年,方法主要是基于蓝-绿波段比值的经验统计法。近些年,随着水色遥感器的改进及数据处理方法的深入研究,提出了荧光高度法[2]和神经网络法[3-5]。本文基于SeaBAM(SeaWiFSBio-OpticalAlgorithmMini-Workshop)小组搜集的全球范围叶绿素浓度与离水辐射率的同步观测数据,利用神经网络方法反演海水叶绿素浓度,并将其结果与SeaBAM经验算法进行了比较及分析。     

马氏珍珠贝肉风味食品开发研究

研究了马氏珍珠贝肉制作五香、蒜香、姜汁三种风味即食制品的加工工艺技术。采用幅照保鲜技术对风味制品进行灭菌防霉处理,产品的保质期达到 １８０ ｄ 以上。对风味制品进行营养成分分析,结果表明：马氏珍珠贝肉风味制品是一种高蛋白、低脂肪、营养价值丰富的海产品,并富含牛磺酸（３．９ ｍ ｇ／ｇ）、多不饱和脂肪酸（ＥＰＡ＋ ＤＨＡ 占脂肪总量 ３４．７％ ）和多种无机质、微量元素和维生素