SERUM IMMUNOGLOBULIN OF THE MANDARIN FISH,SINIPERCA CHUATSI WITH DEVELOPMENT OF POLYCLONAL ANTIBODY

Serum immunoglobulin from the mandarin fish,or the so-called Chinese perch,Siniperca chuatsi(Basilewsky),was successfully purified using affinity chromatography.Heavy and light chains were detected on electrophoresis gel,with molecular weights being estimated at 72 and 29 kDa,respectively.The tetrameric IgM of S.chuatsi was calculated to be 808 kDa.The rabbit polyclonal antisera against the purifed immunoglobulin were developed and tested by Westem blot analysis.The antisera reacted strongly with the heavy chains of S.chuatsi immunoglobulin.Humoral immune responses of the mandarin fish can then be examined using the developed polyclonal antibody.     

Effect of germanium dioxide on growth of Spirulina platensis

This study on the effect of different concentrations of germanium dioxide (GeO₂) on the specific growth rate (SGR), pigment contents, protein content and amino acid composition ofSpirulina platensis showed that Ge was not the essential element of this alga; that GeO₂ could speed up growth and raise protein content ofS. platensis, and could possibly influence the photosynthesis system. The concentration range of GeO₂ beneficial to growth ofS. platensis is from 5–100mg/l. GeO₂ is proposed to be utilized to remove contamination byChlorella spp. usually occurring in the cultivation ofSpirulina.     

Length frequency analysis ofPleuragramma antarcticum,Electrona antarctica,Protomyctophum bolini

British Antarctic Survey (BAS) data collected near the Antarctic Peninsula area were used in this 1988/1989 study on the length frequency of three Antarctic fish species (Pleuragramma antarcticum, Electrona antarctica and Protomyctophum bolini) showing that the standard length (SL) ofP. Antarcticum ranged from 26.0 mm to 161.0 mm, that the 26.0 mm to 54.0 mm SL juveniles could be considered to be the age group 1 fish, and that the SL ofE. antarctica ranged from 39.4 mm to 89.7 mm for the males and 48.0 mm to 118.0 mm for the females, which may contain at least 6 age groups. This is more than previously thought.P. Bolini had 29.2 mm to 60.2 mm SL and complicated age composition.     

Seasonal fluctuation of Myxobolus gibelioi（Myxosporea） plasmodia in the gills of the farmed allogyno—genetic gibel carp in China

The seasonal fluctuation of the plasmodia ofMyxobolus gibelioi Wu et Wang, 1982 in the gill filaments of the allogynogenetic gibel carpCarassius auratus gibelio (Bloch) in a fish pond in Hubei Province of China was investigated from August 1999 to July 2000. A total of 445 fish was examined; the overall prevalence of the plasmodium infection in the fish was 64.94% and mean abundance of plasmodia was 11.65±27.85 (mean±SD). Significant seasonal changes in prevalence and mean abundance, with higher levels of the plasmodia infection from late spring to autumn, were observed. This research was financially supported by the Projects for Knowledge Innovation of the Chinese Academy of Sciences (No. KSCX2-1-04 and No. 1999051) and the Science & Technology Commision of Wuhan City (No. 995004095G).     

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.     

Effect of nitrogen and phosphorus on the growth of a red tide dinoflagellateScrippsiella trochoidea (Stein) Loeblich III

Scrippsiella trochoidea (Stein) Loeblich III was grown in a nitrogen-or phosphorus-limited batch culture system in laboratory. Growth rates and cellular Chl-a were measured as functions of nitrate and phosphate concentrations. Growth rates were hyperbolic with both nitrate and phosphate concentration and fit the Monod equation. The minimum cell quota of nitrogen and phosphorus and then the optimum N∶P ratio ofS. trochoidea were estimated in this study. Measurement of phosphate concentration in Jiaozhou Bay suggest that phosphorus is the limiting factor ofS. trochoidea growth. Contribution No. 3679 from the Institute of Oceanology, Chinese Academy of Sciences. Project 39790110 supported by NSFC, the study aslo supported by the National Climbing Project B (PDBG-7-2) and partly supported by an MF grant from Hong Kong University of Science and Technology.     

A study on pathogens of Chinese prawn (Penaeus Chinensis) virus diseases

This pathogenic study shows that the viral diseases of Chinese prawns (Penaeus chinensis, O'sbeck) is due to three kinds of viruses: epithelium envelope baculovirus ofPenaeus chinensis (EEBV-PC, detected by the authors in 1993), infections hypodermal and hematopoietic necrosis virus, and hepatopancreatic parvo-like virus, and that the first two viruses seem to be the main pathogens of the epidemic in the northern regions in 1993.     

Disease resistance and humoral immunomodulatory effects of vitamin C on grouper,Epinephelus awoara

The disease resistance and humoral immunomodulatory effects of vitamin C administered orally to grouper,Epinephelus awoara maintained on a frozen fish diet supplemented with vitamin C at 500, 1000, 1500 and 2000 mg/kg were investigated. After 20 weeks, the growth rates of the groups with high level of vitamin C apparently increased. The untreated fish had symptoms of vitamin C deficiency. The endogenous liver tissue vitamin C levels were found to reflect well the dietary treatments. After intraperitoneal injection or bath challenge with a virulent strain ofVibrio vulnificus, fish fed with high level vitamin C showed significantly higher survival rate compared with the normal control group. Vaccination with formalin inactivatedV. vulnificus significantly enhanced the specific antibody production in fish treated with vitamin C, and completely protected from strong bacterial challenge the groups fed on fish with vitamin C 1500 and 2000 mg/kg diet. Project 39500113 supported by NSFC and also by Guangdong Science Research Foundation (Grant No. 950711).     

Vaccination in three different ways against vibriosis of <Emphasis Type="Italic">Seriola dumerili</Emphasis> caused by <Emphasis Type="Italic">Vibrio hollisae</Emphasis>

Bacterin was prepared by formalin-inactivating the virulent strain of Vibrio hollisae isolated from diseased Seriola dumerili （amberjack） suffering from vibriosis. Healthy S. dumeriIi were vaccinated by respective procedures of intramuscular injection, immersion, and orally administration. Results of the three different vaccinations were compared. Blood was drawn from the vaccinated fish every 7 days, and the antibody titers and lysozyme activities of the sera were determined. The antibody titer of injected fish was 1：40 at 7 d, and reached its peak of 1：320 at 28 d, while the fish vaccinated by immersion and orally administration exhibited weak antibody responses, the antibody titres of 〈1：10, 1：20, 1：160 were observed at 7 d, 14 d, 35 d respectively. Compared with the control, the vaccinated fish exhibited significantly higher lysozyme activities （P〈0.05）. Upon challenge with virulent strain, the relative percent survival （RPS） of injected, immersed and oral administrated fish were 75%, 45%, and 40% respectively, and the injected fish showed significantly higher RPS than immersed and oral administrated fish. The results suggested that vaccination of S. dumerili by the injection would be the best strategy to prevent the vibriosis in S. dumerili farm.     

Amino acid composition and functional properties of giant red sea cucumber (Parastichopus californicus) collagen hydrolysates

Giant red sea cucumber (Parastichopus californicus) is an under-utilized species due to its high tendency to autolysis.The aim of this study was to evaluate the functional properties of collagen hydrolysates from this species.The degree of hydrolysis (DH),amino acid composition,SDS-PAGE,emulsion activity index (EAI),emulsion stability index (ESI),foam expansion (FE),and foam stability (FS) of hydrolysates were investigated.The effects of pH on the EAI,ESI FE and FS of hydrolysates were also inves-tigated.The results indicated that the β and α1 chains of the collagen were effectively hydrolyzed by trypsin at 50℃ with an En-zyme/Substrate (E/S) ration of 1:20 (w:w).The DH of collagen was up to 17.3% after 3 h hydrolysis with trypsin.The hydrolysates had a molecular weight distribution of 1.1 17 kDa,and were abundant in glycine (Gly),proline (Pro),glutamic acid (Glu),alanine (Ala) and hydroxyproline (Hyp) residues.The hydrolysates were fractionated into three fractions (< 3 kDa,3 10 kDa,and > 10 kDa),and the fraction of 3 10 kDa exhibited a higher EAI value than the fraction of > 10 kDa (P<0.05).The fraction of > 10 kDa had higher FE and FS values than other fractions (P<0.05).The pH had an important effect on the EAI,ESI,FE and FS.All the fractions showed undesirable emulsion and forming properties at pH 4.0.Under pH 7.0 and pH 10.0,the 3 10 kDa fraction showed higher EAI value and the fraction of > 10 kDa showed higher FE value,respectively.They are hoped to be utilized as functional ingredients in food and nutraceutical industries.     

Effects of fishes on the growth and survival of Chinese mitten crab,Eriocheir sinensis

This experiment on the effects of fishes (Pelteobagrus fulvidraco, Channa argus, Cyprinus carpio) on growth and survival of juvenile Chinese mitten crab,Eriocheir sinensis was conducted in cages placed in Bao'an Lake. T-tests showed that effects of fishes on the carapace width growth were not significant as compared with the control (no fish). But the effects of these fishes on crab survival were negative. The causes of negative effects of fishes on crab survival were simply discussed on the basis of food habits. This research was financially supported by the Department of Agriculture (96-008-02-03), P. R. China, and Chinese Academy of Sciences (Key Project: KZ951-A1-102).     

ENCLOSURE EXPERIMENTS ON AND LACUSTRINE PRACTICE FOR ELIMINATING MICROCYSTIS BLOOM

Microcystis bloom, one of the most objectionable characteristics of eutrophication in tropical and subtropical waters, occurred in Donghu Lake (East lake) of Wuhan every summer from the 1970s up to 1984, but from 1985 up to now failed to occur there. The cause of its disappearance rema-ined in obscurity until recently. In situ enclosure experiments in the lake for three years showed that the stocking of the filter-feeding silver carp ( Hypophthalmichthys molitrix ) and big-head carp (Aristichthys nobilis ) played a decisive role in eliminating Microcystis bloom from the lake; but that recurrence of the bloom is possible under certain conditions. This paper presents the details and the results of enclosure ex-periments. The authors‘ analysis of fish biomass data obtained by echo-sounding and the fishery produc-tion of the lake over the years, revealed that the recurrence of Microcystis bloom can be prevented so long as the combined biomnss of silver carp and big-head carp remains at or exceeds 50 g per cubic meter of lakewater, as was the case in the lake‘s 1985 fish yield of 1015 t.     

Identification and characterization of a new IgE-binding protein in mackerel (Scomber japonicus) by MALDI-TOF-MS

As fish is one source of the ‘big eight’ food allergens,the prevalence of fish allergy has increased over the past few years.In order to better understand fish allergy,it is necessary to identify fish allergens.Based on the sera from fish-allergenic patients,a 28 kDa protein from local mackerel (Scomber japonicus),which has not been reported as a fish allergen,was found to be reactive with most of the patients’ sera.The 28 kDa protein was analyzed by MALDI-TOF-MS (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry).Mascot search in NCBI database (Date: 08/07/2010) showed that the top protein matched,i.e.triosephosphate isomerase (TPI) from Xiphophorus maculatus and Poecilia reticulata,had a mowse (molecular weight search) score of 98.In addition,TPI from Epinephelus coioides also matched this mackerel protein with a mowse score of 96.Because TPI is con-sidered as an allergen in other non-fish organisms,such as lychee,wheat,latex,archaeopotamobius (Archaeopotamobius sibiriensis) and crangon (Crangon crangon),we consider that it may also be an allergen in mackerel.     

FOOD SOURCES AND CARBON BUDGET OF CHINESE PRAWN PENAEUS CHINENSIS

This study deals with contribution of artificial food pellet and natural food to Chinese prawn (Penaeus orientalis) growth in a semiintensive culture pond. The prawn carbon consumption, budget, and the effects of some factors on the budget were investigated. The results showed that 26.2% of P. orientalis growth carbon came from formulated feed at the initial culture stage (when the prawns were 0.06±0.01 g in wet weight), and was 62.5% when the prawns were 9.56±1.04 g. The remaining part of the growth carbon was derived from organic fertilizer and natural food. The highest growth rate occurred at 20×10-3 salinity. Suitable salinity for culturing Chinese prawn was (20-28)×10-3.     

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).     

Effects of blue light on gametophyte development of Laminaria japonica (Laminariales, Phaeophyta)

Laminaria gametophyte was greatly influenced by light in its growth and development. Using light-emitting diodes (LED) as blue and red light sources, we analyzed the light effect on gametophytes development ofLaminaria japonica Aresch. The gametophytes were obtained from zoospores collected in April, May, July, 2003 and September, 2004. We found that the growth of gametophytes was stimulated by increasing intensity of blue light (BL) and red light (RL) illumination, of which BL was obviously stronger than that of RL. The fertilization of gametophytes depended largely on BL, and only sufficient BL illumination could take the reproductive effect. In addition, we noticed that there was a significant difference in light responses for gametophytes developed from zoospore collected in different times. For zoospores released in April, under BL₁ (73.90 μmol photons/m·s), the unicellular female gametophytes and multi-cellular male gametophytes produced eggs and sperms respectively, and further developed towards sporophytes. However, for gametophytes developed in May, July or September, they became multi-cellular and never formed oogonia or antheridia. It is believed that theLaminaria sporangium maturation stage could affect the gametophytes reaction to BL under laboratory culture conditions. Therefore, cryptochrome- or phototropin-like BL photoreceptors is probably involved in BL-induced development ofLaminaria gametophytes. This research was supported by the NSFC (No.40376049) and Japan Society for the Promotion of Science (The Invitation Fellowship for Research in Japan 2002).     

Appearance pattern of tooth germs with developmental process inMylopharyngodon piceus

The pharyngeal dental formula ofMylopharyngodon piceus is 4–5 as a rule, and the dentition is asymmetrical. It is difficult to identify each tooth in the larval dentition. In this paper the appearance pattern of tooth germ with development process in this fish is described in detail. The formation pattern of the left dentition is contrasted with that of the right one. In the developmental process, the left pharyngeal dentition lacks teeth at position An3. Thus the left dentition is D-type as designated by Nakajima (1984), while the right one is A-type. This project was funded by the International Cooperation of Japan-China and the National Natural Science Foundation of China.     

Use of rapd for detecting and identifyingPorphyra (Bangiales, Rhodophyta)

Randomly amplified polymorphic DNA (RAPD) technique was applied to assess the genetic variations and phylogenetic relationships in 4 species ofPorphyra. The samples were collected from the coast of Canada, Vietnam, Zhoushan, Fujian and Qingdao in China. Amplifications with 20 primers were carried out under predetermined optimal reaction conditions (samples were first heated at 94°C for 5 min. and followed by 45 cycles of 1 min at 94°C, 1 min at 36°C, and 2 min at 72°C, then held at 72°C for 10 min). The amplified products were scored as present (1) or absent (0) for each DNA sample and an index of genetic similarity (F) was calculated by using Nei & Li’s matching coefficient method (1979). The value of (1−F) was used to quantify the genetic distances between species and construct a phylogenetic tree. The relationship indicated by the UPGMA and NJ cluster analysis on the values of the genetic distance is in good overall agreement with classical taxonomy. The obvious differences between natural and cultivated population ofP. haitanensi suggest that variation or hybridization with other species occurred during the culture. Contribution No. 100 from the Experimental Marine Biology Laboratory, IOCAS. Contribution No. 3289 from the Institute of Oceanology, Chinese Academy of Sciences. The research supported by the Foundation of the Chinese Academy of Sciences.