The induction and extraction of metallothioneins inArtemia

A small amount of heavy metal binding protein, identified by BioRad Protein Assay, has been isolated from the adult brine shrimp,Artemia franciscanus. This protein has an apparent molecular weight between 6000 to 9000 dalton. a UV absorption peak at 260 instead of 280 nm like most proteins; and has high affinity towards binding with radioactive labeled¹⁰⁹Cd. These characteristics are similar to that of metallothioneins reported for many vertebrate and invertebrate, marine and terrestrial animals. After the brine shrimp is exposed to a small amount of Cd²⁺ for 24 h, a large amount of metallothionein can be isolated, showing the inducibility of this detoxifying protein in the adultArtemia in a short period of time.     

Effect of carbon and nitrogen ratio control on Artemia growth,water quality,biofloc microbial diversity under high salinity and zero-water exchange culture condition

Biofloc technology has been applied successfully in the intensive aquaculture of several fish and shrimp species.The growth of heterotrophic microorganisms can be stimulated through adding extra carbon,which reduces the nitrogen level in the water and provides microbial protein to the animals.However,most of the studies and practical applications have been conducted in freshwater and marine environment.This paper focused on brine shrimp Artemia that lives in high salinity environment together with other halophilic or halotolerant microorganisms.The effect of carbon supplementation on Artemia growth,water quality,and microbial diversity of bioflocs was studied in the closed culture condition without any water exchange.The salinity of the culture medium was 100.A 24-d culture trial was conducted through supplementing sucrose at carbon/nitrogen(C/N)ratio of 5,15,and 30(Su5,Sul5,and Su30),respectively.The culture without adding sucrose was used as a control.Artemia was fed formulated feed at a feeding ration of 60% recommended feeding level.The results showed that sucrose supplementation at higher C/N ratio(15 and 30)significantly improved the Artemia survival,growth and water quality(P0.05).Addition of sucrose at C/N ratio of 15 and 30 significantly increased biofloc volume(BFV)(P0.05).The Illumina MiSeq sequencing analysis showed that supplementing carbon at C/N ratio of 15 had a better total bacterial diversity and richness,and shaped the microbial composition at genera level.This study should provide information for studying the mechanism of biofloc technology and its application in high salinity culture conditions.     

The brine shrimp (Artemia parthenogenetica) as encapsulation organism for prophylactic chemotherapy of fish and prawn

Brine shrimp (Artemia parthenogenetica) which had ingested three water-insoluble antibacterial drugs i.e. sulfadiazine(SD), oxytetracycline (OTC) and erythromycin estolate (ERY-Es) were fed toTilapia and Mysis IIIof Penaeus orientalis K. The drug contents in the predators were then determined. After administration of drugs toTilapia and Mysis III, through the bio-encapsulation of the brine shrimp, efficacious therapeutical concentration of OTC and ERY-Es (but not SD) in the predators could be reached and maintained for more than 8 hours.     

EFFECT OF Cu, Cd AND Cu:Cd MIXTURE ON THE BIOCHEMICAL COMPOSITION OF TWO PENAEID SHRIMP POSTLARVAE

IwrRODUcrIONBiochewhcalindiasrnaybeusedtoreboresponsestospedccontandnantSincludingheaVyrnetaIs.ChineseshrimpfisheriesarebasedonpenaeidsofWhichPenaeuspoiciIlatusistheddrinantsthe(Xuetal.,l993)incoastalwatersofFujianProvince.Larvalpenaeids(Andelsonetal.,l976)likeotherplanktoniccrustaonns(Clarkl992),arepartioulariysensitivetoheaVymetalcontandnatedwaterduetothefortnationofmetalbindingproteinsintheirbodies.Cytoplasndcproteins,dondnatedbythestilphydrygrouphave~reportedinvariousdassesofphylog…     

Dormant stages of crustaceans as a mechanism of propagation in the extreme and unpredictable environment in the Crimean hypersaline lakes

A pool of dormant stages of planktonic organisms in saline lakes is a substantial component in the plankton communities;we need to take it into account to understand plankton dynamics.Hypersaline water bodies in Crimea,the largest peninsula in the Black Sea,constitute a very characteristic and peculiar habitat type in the region.We examined the presence of crustacean resting stages in sediments of dried up sites of the Crimean hypersaline lakes.Sediment samples were taken in 9 different lakes.Experiments performed on the hatching of these resting stages showed the presence of Moina salina(Cladocera),parthenogenetic Artemia and Artemia urmiana(Anostraca),Eucypris mareotica( inflata)(Ostracoda),and Cletocamptus retrogressus(Harpacticoida).Comparing the experimental results obtained with clean dried brine shrimp cysts and those kept in sediment samples,it was noted that clean cysts hatched much faster than those from sediments did.Some components in bottom sediments slow down and desynchronize hatching from resting eggs in different groups of crustaceans.The sediments of different lakes inhibited the nauplii output from Artemia and ostracod resting eggs to different degrees.More data are needed before we can discuss the reasons of this inhibition.The nonsynchronous output of active stages from the bottom resting ones may be an adaptation that allows crustacean species to exist in extreme and unpredictably changing environments,avoiding the risk that all may emerge at once under unsuitable conditions.     

A Study on the Contribution of Different Food Sources to Shrimp Growth in an Intensive Fenneropenaeus chinensis Pond

Stable isotope methods can be used to determine the food sources and prey items of aquatic organisms accurately andreliably.This study examined the relative contribution of artificial foods (the formulated feed and Artemia) and natural foods toshrimp growth in an intensive Fenneropenaeus chinensis pond by using carbon and nitrogen stable isotopes.The results showed that the nutrition utilization efficiency of the harvested shrimp was low,only 33.18% of feed nitrogen and 21.73% of feed carbon beingconverted to shrimp flesh.Our stable isotope results showed that the shrimp obtained nutrition for maximum growth from artificialfoods,whose contribution was 93.5%,with the remaining attributed to the natural foods.However,there was 0.94 t harvested shrimpderived from natural foods (the rest of 13.56t harvested shrimp derived from artificial foods) in lha intensive pond with a shrimpproduction of 14.50 tha-1.Therefore,unit area shrimp production can be increased by increasing the contribution proportion of natu-ral foods in intensive shrimp farming.     

Shrimp arginine kinase being a binding protein of WSSV envelope protein VP31

Viral entry into the host is the earliest stage of infection in the viral life cycle in which attachment proteins play a key role. VP31 (WSV340/WSSV396), an envelope protein of white spot syndrome virus (WSSV), contains an Arg-Gly-Asp (RGD) peptide domain known as a cellular attachment site. At present, the process of VP31 interacting with shrimp host cells has not been explored. Therefore, the VP31 gene was cloned into pET30a (+), expressed in Escherichia coli strain BL21 and purified with immobilized metal ion affinity chromatography. Four gill cellular proteins of shrimp (Fenneropenaeus chinensis) were pulled down by an affinity column coupled with recombinant VP31 (rVP31), and the amino acid sequences were identified with MALDI-TOF/TOF mass spectrometry. Hemocyanin, beta-actin, arginine kinase (AK), and an unknown protein were suggested as the putative VP31 receptor proteins. SDS-PAGE showed that AK is the predominant binding protein of VP31. An i n vitro binding activity experiment indicated that recombinant AK’s (rAK) binding activity with rVP31 is comparable to that with the same amount of WSSV. These results suggested that AK, as a member of the phosphagen kinase family, plays a role in WSSV infection. This is the first evidence showing that AK is a binding protein of VP31. Further studies on this topic will elucidate WSSV infection mechanism in the future.     

南海海域三疣梭子蟹的幼体培育

幼体期Z1和Z2投喂扁藻和轮虫,Z2后期开始以轮虫和卤虫无节幼体混合投喂,Z3后用卤虫无节幼体投喂,大眼幼体用不同饵料培育,进行南海海域三疣梭子蟹幼体培育,结果表明:溞状幼体平均成活率为15.3%;Z2变态为Z3、Z4变态为M为溞状幼体培育的敏感期,幼体的阶段成活率较低;以卤虫无节幼体、冰冻卤虫成虫、活卤虫成虫为饵料,均能使培育大眼幼体至幼蟹,其中活卤虫成虫投喂、加防残网片培育效果最好,成活率达80%。     

Influence of Ultrasonic Treatment on the Allergenic Properties of Shrimp （Penaeus vannamei ） Allergen

The present study was undertaken to determine whether high intensity ultrasound could reduce the allergic properties of shrimp allergens. Reducing the allergenic properties of these allergens will be beneficial to allergic individuals. Samples of shrimp protein extract and shrimp muscle were treated by high-intensity ultrasound with water bathing at 0 ℃ or 50 ℃ for different time periods. The treated and untreated samples were then analyzed by SDS-PAGE, Western blots and competitive inhibition ELISA （Ci-ELISA） to determine the shrimp allergenicity. The results show that high-intensity ultrasound has no effect on allergenicity when the extracts were treated at 0℃. However, a significant decrease was observed in the level of the major shrimp allergen, Pen a 1, when the samples were treated at 50 ℃. In the determination of allergenicity with CiELISA, a reduction in IgE binding was also observed.     

黄河三角洲高效生态经济区卤水资源开采潜力评价

黄河三角洲高效生态经济区卤水资源较为丰富,历经几十年的集中开采后,已出现卤水开采水位下降、卤度及有益离子含量下降等问题,该文在核算研究区卤水资源量的基础上,结合开发利用现状进行了开采潜力评价,以期为当地卤水资源开采规划提供科学依据。     

Negative effects of Phaeocystis globosa on microalgae

The potential allelopathic effects of the microalga, Phaeocystis globosa Scherffel, on three harmful bloom algae, Prorocentrum donghaiense Lu, Chattonella marina (Subrahmanyan) Hara et Chihara and Chattonella ovata Hara et Chihara were studied. The growth of C. marina and C. ovata was markedly reduced when the organisms were co-cultured with P. globosa or cultured in cell-free spent medium. Haemolytic extracts from P. globosa cells in the senescence phase had a similar inhibitory effect on the three harmful bloom algae. However, P. globosa had less influence on the brine shrimp, Artemia salina. These results indicate that P. globosa may have an allelopathic effect on microalgae, which would explain the superior competitive abilities of P. globosa. Because the addition of the haemolytic toxins from P. globosa had similar effects on algae as spent media, these compounds may be involved in the allelopathic action of P. globosa.     

莱州湾南岸地下卤水水位动态变化与开采量关系研究

由于多年来大量开采地下卤水,莱州湾南岸已经形成多个地下卤水降落漏斗.该文根据区内盐场水位长期观测资料,较系统地分析了地下卤水水位动态变化规律,查明了卤水水位与开采量的关系,建立了区内卤水水位与开采量相关关系方程.通过分析说明开采对水位的影响,为改善区内地质环境,合理利用卤水资源提供了科学依据.     

不同养殖模式虾池弧菌对抗菌药物的耐药性与虾池水质评价

研究了湛江东海岛3种不同养殖模式虾池弧菌对抗菌药物的耐药性,并对虾池水域环境的水温、盐度、DO、pH、叶绿素a、DOC、无机氮、无机磷、硅酸盐、细菌总数和弧菌数量变化进行了监测和评价。结果表明,调查的三个虾池的弧菌对万古霉素产生耐药性,高位新池和传统精养池的弧菌对利福平产生耐药性,对其他抗菌药尚未形成耐性,3个虾池的弧菌对恩诺沙星和氯霉素高度敏感,传统精养池和天然虾池的弧菌对复方新诺明、庆大霉素、TMP和环丙沙星高度敏感;在水平差异上,高位新池的弧菌比传统精养池和天然虾池对12种抗菌药更具耐药性,同时多重耐药菌株在高位新池中出现,可能与虾苗来源和配合饲料中添加了抗菌药物有关。水质监测结果表明,养殖过程中,虾池水域环境逐渐富营养化,无机氮和Chla超过富营养化阈值,细菌总数增加;两个精养池的水体富营养化程度高于天然虾池;调查期间的弧菌数量在对虾发病的感染剂量阈值范围之内,尚无弧菌病害发生的潜在危险。     

The forming-storing environment and abundance conditions of quaternary underground brine in the embayed coast of Qingdao

ImODUcrIONGroundwaterwthAnneralimtionofmorethan5OgA(about5"Be')"iscalledunder-groundbrine.ltbelongstoevaporationoredepositinliquidstateandisantwortantrnaterialsourceforsaltonkingandthechewhcalindustry.QuatemarylittoralfadesundergroundbrinewasstoredinthelittoralsoneandhadtwobasictypesofgenesisenvironmentflittoralplaincoastenvironmentcharacterindbythecoastalongLaichouBay,andembayedcoastenvironmentcharaCterindbythecoastalzoneofQingdao.TheauthorresearchedthebrineintheLaizhoubaycoastareainthe…     

A Successful Microbound Diet for the Larval Culture of Chinese Shrimp Fenneropenaeus chinensis

A 13-day feeding trial was carried out to evaluate the effectiveness of a microbound diet for rearing the larvae of Chinese shrimp Fenneropenaeus chinensis in comparison with the live foods that consist of Isochrysis galbana, Chlorella vulgaris, Tetraselrnis chuii, rotifer （Brachionus plicatilis） and Arternia sp. Larvae of 0 to 13d post-hatch （dph） were reared in a temperature-controlled semi-open culture system and stocked at a density of 100 larvae L^-1 in tanks, each containing 50 L sterilized seawater with salinity 30-32. Larvae were manually fed either the live foods or the microbound diet 6 times per day. At 13 dph, the growth of the larvae fed on the microbound diet was approximately 84% of that fed on the live foods. The survival rate of the larvae fed on the microbound diet was 44.29% at 13dph, which was not significantly different from that of larvae fed on live foods （63.55%）. The body length and development index （DI） of the larvae fed on the microbound diet were always lower than those of larvae fed on live foods. However, the differences reached significant levels only at 11 and 13 dph （P〈 0.05）. The mean dry weight loss of the microbound diet was 9.2% after 90 min immersion in seawater, indicating that this diet has a good water stability. The microbound diet contains 52.23% crude protein and 10.27% lipid and is easy to prepare. These characteristics of the diet suggest good potentials for its successful use in the larviculture of other penaeid and fish species.     

How can saline and hypersaline lakes contribute to aquaculture development? A review

A considerable part of the world's population is currently experiencing a severe scarcity of freshwater and nutrition. Inland aquaculture has the fastest growth in fresh waters, and this contributes to the eutrophication of freshwater bodies. The increase in freshwater aquaculture impacts on the increasing demand for fresh water. A way to overcome this is to develop aquaculture in saline lakes. This article discusses how saline and hypersaline lakes may contribute to overcome this problem and gives a list of fish and shrimp species that can be cultivated in saline lakes. Successful development of aquaculture depends on a healthy cultured stock of commercial fish and shrimps. A sustainable healthy stock of fish and shrimps can be only maintained using live food for the cultured fish larvae, fry and fingerlings. As well as Artemia spp. there are many other crustacean species with the potential for growing in hypersaline waters. At least 26 copepod species around the world can live at a salinity above 100 g/L with 12 species at a salinity higher than 200 g/L, and these all have excellent nutritional value. There is a high potential to use eukaryotic organisms of diff erent taxa in saline/hypersaline aquaculture for food, agri-aquaculture, diff erent industries and as food supplements.     

Dietary manganese requirement ofP. Vannamei

Graded levels of manganese were supplemented to a semi-purified diet containing 45% crude protein, to provide six levels of manganese (i. e. containing 5, 25, 50, 70, 140 and 210×10⁻⁶, respectively) for two experiments with these experimental diets. The initial weight of shrimp used in the 35–day experiment I was 0.30±0.04 g, and that in the 70–day Experiment II was more than one gram. The results showed that optimum content in the semi-purified diet for the more than 1 gram shrimp ranged from 70 ×10⁻⁶, to 140×10⁻⁶, but supplementation of Mn was not necessary for the small shrimp. Contribution No. 2673 from the Institute of Oceanology, Chinese Academy of Sciences.     

Comparative study on the allergenicity of different Litopenaeus vannamei extract solutions

Allergen extracts are widely used for allergy diagnosis and treatment. The application of shrimp extract is hampered due to the low protein concentration and the inconsistent allergenicity. Extracting solutions are considered to be the primary limiting factor of protein extraction from crustaceans. This study aimed to select an optimal solution for shrimp protein extraction by comparing the allergenicity of different shrimp extracts. The effect of 7 existing or modified extracting solutions were evaluated, including the glycerol-NaCl solution, the glycerol Cocaine's solution, the buffered saline solution, the Cocaine's solution, the Glucose leaching solution, 1 mol L-1 KCl solution, and 0.01 mol L-1 phosphate buffered saline solution with and without dithiothreitolor(DTT). The quantitative(protein concentration) and qualitative parameters(SDS-PAGE protein patterns and immuno-reactivity) were determined using the sodium dodecyl sulfate polyacrylamide gel electrophoresis, enzyme linked immunosorbent assay and immunoblotting assay. Results showed that the 1 mol L-1 KCl solution with DTT was optimal for shrimp protein extraction, which yielded high concentration and allergenicity in the protein extract, including major and minor allergens. The 1 mol L-1 KCl solution with DDT is proposed for preparation of shrimp extract and associated allergy diagnosis, as well as potential applications for other crustaceans.     

Isolation and identification of a bacterium from marine shrimp digestive tract: A new degrader of starch and protein

It is a practical approach to select candidate probiotic bacterial stains on the basis of their special traits. Production of digestive enzyme was used as a trait to select a candidate probiotic bacterial strain in this study. In order to select a bacterium with the ability to degrade both starch and protein, an ideal bacterial strain STE was isolated from marine shrimp (Litopenaeus vannamei) intestines by using multiple selective media.The selected isolate STE was identified on the basis of its morphological, physiological,and biochemical characteristics as well as molecular analyses. Results of degradation experiments confirmed the ability of the selected isolate to degrade both starch and casein. The isolate STE was aerobic, Gram-negative, rod-shaped, motile and non-spore-forming, and had catalase and oxidase activities but no glucose fermentation activity. Among the tested carbon/nitrogen sources, only Tween40, alanyl-glycine, aspartyl-glycine, and glycyl-l-glutamic acid were utilized by the isolate STE. Results of homology comparison analyses of the 16S rDNA sequences showed that the isolate STE had a high similarity to several Pseudoalteromonas species and, in the phylogenetic tree, grouped with P. ruthenica with maximum bootstrap support (100%). In conclusion, the isolate STE was characterized as a novel strain belonging to the genus Pseudoalteromonas.This study provides a further example of a probiotic bacterial strain with specific characteristics isolated from the host gastrointestinal tract.     

芒果粉对真空油炸面包虾裹浆特性及其品质的影响

【目的】研究添加芒果粉对真空油炸面包虾裹浆特性及其品质的影响。【方法】以小麦粉和大豆分离蛋白的裹浆作为对照,研究芒果粉替代部分小麦粉和大豆分离蛋白的裹浆流变学特性;以裹涂率、油脂含量、水分含量、色泽及质构特性为考察指标,研究芒果粉替代部分小麦粉和大豆分离蛋白的裹浆对真空油炸面包虾品质的影响;以最佳芒果粉添加量的真空油炸面包虾为研究对象,测定外裹层黄酮物质含量,并采用羟自由基和DPPH自由基体系对产品外裹层黄酮物质的抗氧化性进行分析。【结果】所有配方的裹浆均表现为触变性,呈现剪切稀化特性,具有假塑性流体特征,并符合幂率流体,芒果粉可显著提升裹浆的黏度;芒果粉可显著增大面包虾裹涂率,改善产品质构和色泽特性,降低产品油脂含量并提升其水分含量(P <0.05)。当芒果粉添加量从10%增加到30%时,面包虾产品油脂质量分数从24.84%降低到20.58%,产品的脆性和色泽中的亮度值(L*)、黄度值(b*)均得到了显著提升(P <0.05)。当小麦粉质量分数66%、大豆分离蛋白质量分数4%、芒果粉质量分数30%时,真空油炸面包虾产品的各项品质最佳;真空油炸面包虾产品外裹层黄酮物质质量分数为(46.75±2.87)mg/100g,其对羟自由基和DPPH自由基的清除效果方面都均强于维生素C。【结论】芒果粉可提升裹浆黏度,改善真空油炸面包虾产品品质,其外裹层较强的抗氧化性对于真空油炸裹涂制品延长货架期具有一定积极作用。