Chromosome behavior of heat shock induced triploid in Fenneropenaeus chinensis

Triploidy was induced in Chinese shrimpFenneropenaeus chinensis by 30±0.5°C heat shock treatment (initiated at 20 min after fertilization) for 10 min to inhibit the release of PB2 at 18.0°C. The highest triploid rate obtained was 84.5% in nauplius stage. The effect of heat shock treatment on meiosis and cleavage of eggs was investigated in this work aimed to establish efficient procedures for triploid induction and to gain understanding of the mechanism of triploid production. Three pronuclei that could be observed in the treated eggs under fluorescence microscope developed into triploid embryos. Some abnormal chromosome behavior was observed in heat shocked eggs. Contribution No. 4407 from the Institute of Oceanology, Chinese Academy of Sciences. This work was supported by Project under Major State Basic Development Program (G1999012009) and Knowledge Innovation Program of the Chinese Academy of Sciences (KZCX 2-211)     

Assessing the germplasm of Laminaria (phaeophyceae) with random amplified polymorphic DNA (RAPD) method

Eighteen gametophytes includingL. japonica, L. ochotensis andL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for final amplification test. Among the total of 205 bands amplified, 181 (88.3%) were polymorphic. The genetic distance among different strains ranged from 0.072 to 0.391. The dendrogram constructed by unweighted pair-group method with arithmetic (UPGMA) method showed that the female and male gametophytes of the same cell lines could be grouped in pairs respectively. It indicated that RAPD analysis could be used not only to distinguish different strains ofLaminaria, but also to distinguish male and female gametophyte within the same cell lines. There is ambiguous systematic relationship if judged merely by the present data. It seems that the use of RAPD marker is limited to elucidation of the phylogenetic relationship among the species ofLaminaria. Contribution No. 4389 from the Institute of Oceanology, Chinese Academy of Sciences. This research was supported by the State's High-tech Project of China, Marine 863 project (No. 819-03-02) and (2001AA621090).     

Tetraploid induction by inhibiting mitosis I in scallopChlamys farreri

Tetraploid induction was carried out by inhibiting mitosis I in fertilized eggs ofChlamys farreri. Mitosis I was blocked with cold shock (5–7°C), Cytochalasin B (0.75 mg/L) and 6-dimethylaminopurine (6-DMAP) (60–75 mg/L) when 60% fertilized eggs released polar body II at 20°C. At 4-cells embryo stage, the ploidy was determined by counting chromosome number. In control groups, most embryos were diploids (72.22%) and aneuploids (24.78%). In Cytochalasin B, cold shock and 6-DMAP treated groups, tetraploids were respectively 10.51%, 4.08%, and 13.34%; aneuploids were 43.10%, 35.93% and 29.16%, and triploids were 7.84%, 8.52% and 18.33%. At D-larva stage, ploidy was determined by flow cytometry (FCM). The ploidy analysis of day 2 larvae showed diploids in control group and also in three treated groups. Juvenile scallops (0.2–0.3cm) which were harvested in two control groups and two CB treated groups were all diploids through checking ploidy individually by FCM. Contribution No. 3648 from the Institute of Oceanology, Chinese Academy of Sciences. Contribution No. 238 from the Experimental Marine Biology Laboratory, Chinese Academy of Sciences. Project funded by grant (No. 819-01-07) from the Chinese Science and Technology Ministry and also by “Hundred Talents Plan” grant from the Chinese Academy of Sciences.     

PHYLOGENETIC RELATIONSHIPS AMONG PUFFERFISH OF GENUS TAKIFUGU BY RAPD ANALYSIS

AmpLifications with 300 10-base primers under predetermined optimal reaction yielded 2762 reproducible amplified fragments ranging from 200 to 3000 bp. Genetic distances among five species of Takifugu and Lagocephalus spadiceus as outgreup were calculated based on the presence/absence of the amplified fragments. Topological phylogenie trees of the 5 species of Takgfugu and the outgroup were gen-erated by Parsimony and Neighbor-Joining analysis based on RAPD data set. The genetic distance between T. rubripes and T. pseudommus was almost the same as that between individuals within each species, but was much smaller than that among T. rubripes, T. pseudommus and the other species. The molecu-lar data from RAPD analysis convincingly showed that T. rubripes and T. pseudommus should be the same species.     

Reproductive morphology ofEucheumagelatinae (Esper) J. Agardh (Solieriaceae,Gigartinales, rhodophyta)

Eucheuma gelatinae (Esper) J. Agardh has vegetative and reproductive features distinguishing it from other species ofEucheuma. The occurrence of reproductive nemathecia containing carpogonial branches, auxiliary cells and post-fertilization stages including gonimoblast and pericarp initiation, fusion cell formation stages and carposporophyte development are described and ilustrated for the first time for this species. Contribution No. 2380 of the Institute of Oceanology, Chinese Academy of Sciences.     

Effects of Body Weight and Water Temperature on Maximum Food Consumption of Juvenile Sebastodes fuscescens （Houttuyn）

Maximum rate of food consumption ( Cmax ) was determined for juvenile Sebastodes fuscescens (Houttuyn) at water temperature of 10, 15, 20 and 25℃. The relationships of Cmax to the body weight (W) at each temperature were described by a power equation: lnCmax = a b lnW. Covariance analysis revealed significant interaction of the temperature and body weight. The relationship of adjusted Cmax to water temperature (T) was described by a quadratic equation: Cmax = -0.369 0.456T - 0.0117T^2. The optimal feeding temperature calculated from this equation was 19.5 ℃. The coefficients of the multiple regression estimation relating Cmax to body weight (W) and water temperature (T) were given in the Table 2.     

STUDY ON EMBRYONIC DEVELOPMENT AND EARLY GROWTH OF TRIPLOID AND GYNOGENETIC DIPLOID LEFI—EYED FLOUNDER, PARALICHTHYS OLIVACEUS （T. et S. ）

The early effects of chromosomal manipulation of eggs and sperm on the yields of trip-loid and gynogenefic diploid larvae of Paralichthys olivaceus were investigated. Triploidy was achieved by cold shocking fen.ilized eggs at 0 - 2℃ for 45 minutes duration 5 minutes after fen.ilization, and the in-duced triploidy rates were 31.2% - 50% and the relative hatching rates were 53.3% - 99%. Gynnge-aetic diploids were obtained when eggs were inseminated with irradiated sperm and cold shocked at 0 -2℃ for 45 minutes duration 5 minutes after fertihzation. The induced gynogenetic diploid rates and the relative hatching rates were 94 % - 96 % and 48.5 % - 68.5 % respectively. The embryonic development of the triploid experimental group and of the gynogenetic diploid experimental group was delayed at first compared with the control group. But from the gastrula stage, it was not delayed anymore. There were no significant differences in the growth of the triploid experimental group larvae and the control group larvae, and in the growth of the gynogenetic diploid experimental group larvae and the control group larvae ac-cording to Student‘s t-test (α = 0.05). The relationship between the early growth of the triploid experi-mental group larvae and that of gynogenetic diploid experimental group larvae was also studied.     

Effect of ration, temperature and body weight on growth of black porgy,Sparus macrocephalus (B.)

Growth of black porgy,Sparus macrocephalus, was studied experimentally at four ration levels from starvation to satiation and four temperatures ranging from 14.8 to 26.8°C. At maximum rations, the specific growth rate (SGR) increased with temperature and was a negative linear function of body weight. The relationship between SGR and ration levels was a decelerating curve. Stepwise regression was used to develop the predictive models for the specific growth rates. Maintenance rations and optimum rations both increased with increased temperature. The relationship between maintenance, optimum rations and body weight was influenced by temperature. Conversion efficiencies increased with ration from zero at maintenance ration to a peak at the optimum ration, then declined with further increased ration. Contribution No. 3127 from the Institute of Oceanology, Chinese Academy of Sciences. Project 39370542 supported by NSFC and study also supported by National Key Laboratory in the Institute of Hydrobiology, Chinese Academy of Sciences.     

Properties of proteolytic toxin of Vibrio anguilolarum from diseased flounder

Extracellular products (ECP) produced byVibrio anguillarum strain M3 originally isolated from diseased flounder (Paralichthys olivaceus) were prepared. ECP of M3 showed gelatinase, casinase, amylase and haemolytic activity on agarose plates. High protease activity against azocasin was detected. Bacterium M2 showed highest growth and protease activity at 25°C. The protease present in ECP showed maximal activity at pH 8 and 55°C; was completely inactivated by application of 80°C heat for 30 min; was completely inhibited by EDTA and HgCl₂, and was partially inhibited by PMSF, SDS, MnCl₂ and iodoacetic acid; but not inhibited by CaCl₂ and MgCl₂. The ECP was toxic to flounder fish at LD₅₀ values of 3.1 μg protein/g body weight. The addition of HgCl₂ and application of heat at 50°C decreased the lethal toxicity of ECP. When heated at 100°C, ECP lethality to flounder was completely inhibited. After intramuscular injection of ECP into flounder, it showed evident histopathological changes including necrosis of muscle, extensive deposition of haemosiderin in the spleen, dilated blood vessels congested with numerious lymphocytes in the liver. These results showed that ECP protease was a lethal factor produced by the bacteriumV. anguillarum M3. Contribution No. 4213 from the Institute of Oceanology, Chinese Academy of Sciences. Funded by projects under the Major State Basic Research Development Program, Grant G1999012003.     

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

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

IMPACT OF STARVATION ON SURVIVAL，MEAT CONDITION AND METABOLISM OF CHLAMYS FARRERI

The effects of 60-day starvation on survival rate. condition index (CI) , changes ofnutrient composition of different tissues, respiration and excretion of scallop ChlartnCs farreri were studiedin laboratory from Oct. 17 to Dec. 15 ,1997. Two groups (control and starvation with 200 individualseach) were cultured in two 2 m3 tanks, with 31 to 32 salinity water at 1 7 “C . Starvation effects were mea-sured alter 1 0, 20, 40 and 60 days. There was no mass mortality of scallops cf the two tanks and survivalrates of the control and starvation groups were 93 .5 % and 92.0 %, respectively. Starvation had strongeffect on the meat condition of the scallops, especially after 10 days; when relative lipid percentagedropped sharply while relative protein percentage increased. The impact of starvation on the oxygen consumption mtc (OCR) and the ammonia-N cxcretion mtc (AER) was obvious.     

Effects of reduced salinity on oxygen consumption and ammonia-N excretion ofChlamys farreri

Effects of reduced salinity on the oxygen consumption rate (OCR) and the ammonia-N excretion rate (AER) of scallopChlamys farreri (3.2–5.9 cm in shell height, 0.147–1.635 g in soft tissue dry weight) were studied in laboratory from March 21, 1997 to April 16, 1997. Under the controlled conditions of reduced salinity from 31.5 to 15.0 and ambient temperature 17°C and 23°C, the concentrations of dissolved oxygen and ammonia-N were determined by the Winkle method and the hypobromite method, respectively. Results showed that with controlled reduced salinity, the mean values of the OCR were 2.17 mg/(g.h) at 17°C, and 2.86 mg/(g.h) at 23°C and that the mean values of the AER were 178.0 μg/(g.h) at 17°C and 147.0 μg/(g.h) at 23°C. The OCR and the AER decreased with reducing salinity from 31.5 to 15.0 both at 17°C and 23°C. The effects of reduced salinity on the OCR and the AER of scallopC. farreri could be represented by the allometric equation and the exponential equation, respectively. Contribution No. 3295 from the Institute of Oceanology, Chinese Academy of Sciences. Project supported by the National Ministry of Science and Technology of China and by the Chinese Academy of Sciences, Grant No. 96-922-02-04 and KZ951-A1-102-02.     

RELATION OF BODY WEIGHT AND FOOD CONSUMPTION TO METABOLIC RATE OF JUVENILE JAPANESE SEA BASS, LATEOLABRAX JAPONICUS

ＩＮＴＲＯＤＵＣＴＩＯＮＴｈｅＪａｐａｎｅｓｅｓｅａｂａｓｓ (ＬａｔｅｏｌａｂｒａｘｊａｐｏｎｉｃｕｓＣ＆Ｖ)ｄｉｓｔｒｉｂｕｔｅｓｗｉｄｅｌｙｉｎｔｈｅｃｏａｓｔａｌｗａｔｅｒｓａｌｏｎｇｔｈｅＢｏｈａｉＳｅａ ,ＹｅｌｌｏｗＳｅａ ,ＥａｓｔＣｈｉｎａＳｅａ,ａｎｄＳｏｕｔｈＣｈｉｎａＳｅａ .Ａｓａｃａｒｎｉｖｏｒｏｕｓｆｉｓｈ ,ｉｔｏｃｃｕｐｉｅｓｔｈｅｈｉｇｈｔｒｏｐｈｉｃｌｅｖｅｌｉｎｔｈｅｍａｒｉｎｅｅｃｏｓｙｓｔｅｍａｎｄｈａｓａｄａｐｔｅｄｔｏｖａｒｉｏｕｓｅｎｖｉｒｏｎｍｅｎｔａｌｒ…     

Development of suspended conchocelis ofPorphyra haitanensis (Bangiales, Rhodophyta)

This Mar. 1993 to Aug. 1994 study on suspended conchocelis ofPorphyra haitanensis showed that there were three patterns for development of vegetative filaments: filaments to filaments by “budding”; filaments to sporangial branchlets by “budding”, or cell swelling. There were also three patterms for sporangial branchlet development: vegetatively propagating, changing into conchospores, or dying. Each developmental stage had one or more different developmental directions between vegetative filaments and sporangial branchlets. Developments from conchosporangial branchlets to conchospores were sequential and irreversible. Although sporangial branchlets formed at 29°C could give rise to filaments, they could not propagate as healthily under the same conditions as those formed at 25°C did. Probably the crucial period of plant cell differentiation is in the late stage of sporangial branchlets. In line with the developmental directions of different stages, the authors regulated the development of conchocelis to get ideal different developmental stages materials to obtain very developmentally homogeneous stages, including filaments and sporangial branchlets. Contribution No 3283 from the Institute of Oceanology, Chinese Academy of Sciences. Study supported by the Climbing Project B PD-B6-4-2 of the State Science and Technology Commission of China.     

REPORT ON PONTONIINE SHRIMPS（CRUSTACEA：DECAPODA）COLLECTED BY JOINT CHINESE—GERMAN MARINE BIOLOGY EXPEDITIONS TO HAINAN ISLAND,SOUTH CHINA SEA i.ANCHISTUS,CONCHODYTES,CORALLIOCARIS

The present paper reports 7 species of pontoniine shrimps collected from Hainan Island,South China Sea,by the Joint Chinese-German Marine Biology Expeditions,in which,Conchodytes nipponensis(De Haan,1844)and Coralliocaris brevirostris Borradaile,1898 are new records from Chinese waters.     

Effect of cytokinins and auxins on the growth of free-living conchocelis ofPorphyra yezoensis

IAA 3-Indolylacetic acid, NAA a-Naphthylacetic acid and cytokinins in PESI culture medium were used in a study on the effects of plant hormones on the growth of free-living conchocelis ofPorphyra yezoensis which showed that its growth in medium with cytokinins, IAA and NAA was more rapid than that in medium with non—phytohormones; that the optimal concentrations for promoting growth were 10 μg/L for IAA and ZA (Zeatin), and 0.1 μg/L for BA 6-Benzyl amino purine and KIN 6-Furfurylamino- purine. Mix use of NAA, IAA and cytokinins, NAA/ZA 1–1000/1 μg/L, NAA/BA 10/1-1000 μg/L, NAA/KIN 1/1-1000 μg/L promoted growth. IAA/ZA 0.1-1/0.1-1 μg/L; IAA/BA 0.1-1/0.1-10 μg/L IAA/KIN 1/0.1-1000 μg/L also promoted growth. Contribution No. 2335 from the Institute of Oceanology, Chinese Academy of Sciences. Contribution No. 94 from the Experimental Marine Biology Lab Qingdao, China. The research was supported by a grant from the Chinese Academy of Sciences.     

Light and electron microscope observations onNephroselmis gaoae sp. nov. (Prasinophyceae)

Nephroselmis gaoae sp. nov. is described on the basis of light and electron microscope observations of cultured material originally collected and isolated from seawater of Jiaozhou Bay, Qingdao, China. The periplasts on the cell body and flagella are covered by five types of scales, two types on the flagella and three on the body. Among these, the morphology and the number of spines of large stellate body scales differ remarkably from those of previously described species ofNephroselmis. Apart from these, the unusual fine structure of the eyespot (stigma) is very characteristic. As in the other species ofNephroselmis, the eyespot lies immediately under the two-membraned chloroplast envelope; unlike the others, however, it is not composed of a number of osmiophilic globules, but consists of about 14 curved rod-shaped osmiophilic bodies arranged loosely and randomly. This feature distinguishes the present new species not only from the other species ofNephroselmis but also from the other motile algal species, the eyespots structure of which had been previously described. Contribution No. 2316 from the Institute of Oceanology, Chinese Academy of Sciences. This paper was presented at the XIVth International Seaweed Symposium held in Brittany, France, 1992.     

A NEW SPECIES OF THE GENUS NEROCILA （ISOPODA： CYMOTHOIDAE） FROM THE EAST CHINA SEA

A new species of cymothoid Isopeda, Nerocila donghaiensis, collected from the body of a sciaenid fish, Johnius sp., is described in the present paper. The new species can be distinguished from the other species of genus Nerocila by its endoped of uropods with deep notch, sharply serrated on the lateral margin and strongly serrated on the medial margin; antennule with 1 st article swollen; dactyli of pereopods Ⅰ, Ⅱ, Ⅳ, Ⅴ swollen.     

Element composition ofSargassum thunbergii

Results of neutron activation analysis (NAA) ofSargassum thumbergii samples collected from the Qingdao Taipingjiao coast on March 20, 1996 showed that (1)Sargassum thunbergii can to some extent accumulate potassium and calcium (the accumulation coefficient was 10); (2) it can accumulate almost all trace elements, especially iron, managanese and zinc; (3) it can accumulate strontium, aluminium thorium, and rare earth elements. Contribution No.3308 from the Institute of Oceanology, Chinese Academy of Sciences. The study supported by the National Ninth-Five-Year Key Project “96-916-04-01”.