Ontogenic study of isozymes in giant tiger prawn,Penaeus monodon (Crustacea: Decapoda)

The patterns of isozyme variation throughout the life cycle of the giant tiger prawn,Penaeus monodon were described for ten enzyme systems (G3PD, MDH, LGG, GPI, MPI, FBP, PGM, LDH, PGD, and PROT). The zymograms showed little variation in the number of isozymic loci expressed during larval development compared with results reported for other penaeid prawns. There were specific ontogenic patterns at some loci. As larvae developed beyond the embryonic stage, there was a tendency for intensity of staining and complexity of banding patterns to increase.     

ON THE ULIRASTRUCTURE AND CLASSIFICATION OF THE HEMOCYTES OF PENAEID SHRIMP, PENAEUS VANNAMEI (CRUSTACEA, DECAPODA)

INTRODUCTIONHemocytesOfcrustaceansplayanimportantroleintheirimmunedefensenetWork(thethall,Cerenius,l992).Thesecellscanremoveforeignpallicleshyphagocytosis(Tyson,Jenldn,l974;Paterson,Stewed,l974)andencaPsulation(HoseandMartin,l989;Perssonetal.,1987)-TheraPidsealingofawoundtopreventhemolYmPhlossanddric~AnsmsfromPenetraingthewoundisanotherimportantprocessiflaqUaiccrustaceansandisthoughttobecormectedwiththeactivitiesofhemocytes(tevin,l985)-AttemPtshadbeenInadefromoverl(X)yearstoclass@…     

Studies on immunodiagnosis of hepatopancreatic parvo-like virus disease of the Chinese penaeid,Penaeus Chinensis I. Purification of the virus

The viral discase of penaeids is indistinet but highly dangerous because their symptoms are often masked by some secondary ones. In order to gain knowledge on how to prevent the occurrence and spread of this viral disease, diagnostic studies on the early phase of the hepatopancreatic parvo-like viral (HPV) disease of culturedPenaeus chinensis was conducted using ummuno-serological techniques. The purification of HPV was successfully done by density gradient ultracentrifugation of cane sugar.     

Effects of hypoxia on dopamine concentration and the immune response of White Shrimp (Litopenaeus vannamei)

Effects of hypoxia on the dopamine concentration and the immune response of White Shrimp Litopenaeus vannamei were studied. The results showed that hypoxia had significant effects on the concentration of dopamine (DA) in the haemolymph, haemocyte count, phenoloxidase activity, phagocytic activity of haemocytes and bacteriolytic and antibacterial activity in the haemolymph (P<0.05). The concentration of the dopamine in haemolymph reached its maximum in the 3.0 and 1.5 mg L⁻¹ DO groups at 12 h and 6 h, and then returned to normal after 24 h and 12 h, respectively. All immune parameters decreased with the reduction of dissolved oxygen. Total haemocyte count (THC), the hyaline cells and semi-granular cells in the 3.0 mg L⁻¹ DO group became stable after 12 h, while granular cells did so after 24 h. The THC and different haemocyte count (DHC) in the 1.5 mg L⁻¹ DO group became stable after 24 h. Phenoloxidase activity and bacteriolytic activity in the 3.0 and 1.5 mg L⁻¹ DO groups reached their stable levels after 24 h and 12 h respectively, while phagocytic activity and antibacterial activity became stable after 24 and 12, and 36 and 24 h, respectively. It was also indicated that the changes of dopamine concentrations in haemolymph, haemocyte count and phenoloxidase activity were obviously related to the exposure time under hypoxic conditions.     

Hematological changes in white spot syndrome virus-infected shrimp, Fenneropenaeus chinensis (Osbeck)

The pathological changes of hemocytes in the haemolymph and hepatopancreas were examined in experimentally and naturally WSSV (white spot syndrome virus) infected Fenneropenaeus chinensis. The results showed that the pathological manifestations of hemocytes were similar among moribund shrimps infected via injection, feeding and by nature. Firstly, the total hemocyte counts (THCs) in WSSV-infected shrimp were significantly lower than those in healthy shrimp. Secondly, necrotic, broken and disintegrated cells were often observed, and a typical hematolysis was present in the haemolymph smear of WSSV-infected shrimp. Thirdly, necrosis and typical apoptosis of hemocytes were detected with TEM in the peripheral haemolymph of WSSV-infected shrimp. Hyalinocytes and semi-granulocytes with masses of WSSVs in their nuclei often appeared, whereas no granular hemocytes with WSSV were found in the hepatopancreas of moribund infected shrimps. All our results supported that hemocytes were the main target cells of WSSV, and hyalinocytes and semigranular hemocytes seemed to be more favorable for WSSV infection in F. chinensis.     

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.     

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.     

Effects of dietary vitamin C on the immune function of shrimps, Penaeus chinensis

Prepared in this experiment were six groups of diets, i.e. VC0, VC1, VC2, VC3, VC4 and VC5 with the contents of vitamin C (VCmg(100 g)⁻¹ diet) of 0, 100, 200, 400, 800, and 1200 respectively. It was found that vitamin C increased the concentrations of immunoglobulin-like (IgG-like, IgA-like and IgM-like) substances in the serum of Penaeus chinensis after a feeding period of 3 weeks. The differences among groups were significant (P<0.01), but there was no difference in the contents of complement3-like and complement4-like substances in the serum (P>0.05). Phenoloxidase (PO) activity in the serum of VC3 group shrimps was higher than that of VC0 and other groups, but no significant difference was observed between VC0 group and other groups. Furthermore, bactericidal activity of the serum to Vibrio parahaemolyticus in shrimps fed with the VC1 diet was higher than that in the other groups (P<0.01), while no difference was demonstrated among all groups for the bactericidal activity to Vibrio alginolyticus (P>0.05). It is, therefore, suggested that vitamin C (100–400 mg(100 g)⁻¹ diets) could be used as an immunostimulant of P. chinensis.     

The ultrastructure of separated and cultured cell of Porphyra yezoensis

There are many reports that cells (protoplasts) separated from the thallus ofPorphyra by enzyme can develop to normal leafy thalli in the same way as monospores. But there are few investigations on the subcellular structure of the isolated vegetative cell for comparison with the subcellular structure of monospores. To clarify whether the separated and cultured cells undergo the same or similar ultrastructure changes during culture and germination as monospores undergo in their formation and germination, we observed their ultrastructure, compared them with those of the monospore and found that the ultrastructure of separated and cultured cells did not have the characteristic feature as that of monospore formation, such as production of small and large fibrous vesicles, but was accompanied by vacuolation and starch mobilization like that in monospore germination. The paper also discusses the relations between monospores and separated and cultured cells. Contribution No. 3875 from the Institute of Oceanology, Chinese Academy of Sciences. Project 39770593 supported by NSFC; Project 96-C01-05-01 of the 9th Five-Year Plan supported by Science and Technology Commision of China.     

Effect of dopamine injection on the hemocyte count and prophenoloxidase system of the white shrimp Litopenaeus vannamei

Effects of dopamine injection on the hemocyte count, phenoloxidase activity, serine proteinase activity, proteinase in-hibitor activity and α2-macroglobulin-like activity in L. vannamei were studied. Results showed that dopamine injection resulted in a significant effect on the parameters measured (P < 0.05), while no significant difference was observed in the control group (0.85% NaCl). In the experimental groups,the hemocyte count reached the minimum in 3 h;granular and semi-granular cells became stable after 12 h and hyaline cells and the total hemocyte count became stable after 18 h. Phenoloxidase activity reached the minimum in 6 h, and then became stable after 9 h. Serine protease activity and proteinase inhibitor activity reached the minimum in 3 h, and α2-macro-globulin-like activity reached the maximum in 3 h,and all the three parameters became stable after 12 h. The results suggest that the activating mechanisms of the proPO system triggered by dopamine are different from those triggered by invasive agents or sponta-neously activated under a normal physical condition.     

Impact of Vibrio parahaemolyticus and white spot syndrome virus (WSSV) co-infection on survival of penaeid shrimp Litopenaeus vannamei

White spot syndrome virus(WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacterial cross or superimposed infections may induce higher shrimp mortality. We used a feeding method to infect L itopenaeus vannamei with WSSV and then injected a low dose of V. parahaemolyticus(WSSV+Vp), or we fi rst infected L. vannamei with a low-dose injection of V. parahaemolyticus and then fed the shrimp WSSV to achieve viral infection(Vp+WSSV). The effect of V. parahaemolyticus and WSSV co-infection on survival of L. vannamei was evaluated by comparing cumulative mortality rates between experimental and control groups. We also spread L. vannamei hemolymph on thiosulfate citrate bile salt sucrose agar plates to determine the number of V ibrio, and the WSSV copy number in L. vannamei gills was determined using an absolute quantitative polymerase chain reaction(PCR) method. L v My D88 and Lvakt gene expression levels were detected in gills of L. vannamei by real-time PCR to determine the cause of the different mortality rates. Our results show that(1) the cumulative mortality rate of L. vannamei in the WSSV+Vp group reached 100% on day 10 after WSSV infection, whereas the cumulative mortality rate of L. vannamei in the Vp+WSSV group and the WSSV-alone control group approached 100% on days 11 and 13 of infection;(2) the number of Vibrio in the L. vannamei group infected with V. parahaemolyticus alone declined gradually, whereas the other groups showed signifi cant increases in the numbers of Vibrio( P 0.05);(3) the WSSV copy numbers in the gills of the WSSV+Vp, Vp+WSSV, and the WSSV-alone groups increased from 10 5 to 10 7 /mg tissue 72, 96, and 144 h after infection, respectively. These results suggest that V. parahaemolyticus infection accelerated proliferation of WSSV in L. vannamei and vice versa. The combined accelerated proliferation of both V. parahaemolyticus and WSSV led to massive death of L. vannamei.     

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.     

Effects of heavy metals （Pb^2＋ and Cd^2＋） on the ultrastructure, growth and pigment contents of the unicellular cyanobacterium Synechocystis sp. PCC 6803

The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb²⁺ and Cd²⁺ (0, 0.5, 1, 2, 4, 6 and 8 mg/L). The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased (>4 mg/L Pb²⁺) metal concentration. The photosynthetic apparatus (thylakoid membranes) were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration (8 mg/L Pb²⁺), the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations (0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L Pb²⁺), both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents (chlorophyll α, β carotene and phycocyanin) were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb²⁺, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll α, β carotene and phycocyanin respectively over the control. Corresponding reductions for the same Cd²⁺concentrations were 57.83, 48.94 and 56.90%. Lethal concentration (96 h LC₅₀) values (3.47 mg/L Cd²⁺ and 12.11 mg/L Pb²⁺) indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd²⁺ than Pb²⁺. Supported by the Chinese Scholarship Council     

Eualus heterodactylus sp. nov., a new hippolytid shrimp from Chinese coast of the Yellow Sea （Crustacea,Decapoda, Caridea）

A new species from the caridean family Hippolytidae, Eualus heterodactylus sp. nov. is described and illustrated based on the specimens collected from Chinese coast of the Yellow Sea. The new species is a part of an informal species group characterized by the possession of epipods on the anterior three pairs of pereopods, and is distinguished from other species of this group by the dactyli of the third to fifth pereopods possessing distinctly stair-like flexor margins in males.     

Detection of white spot syndrome virus (WSSV) ofPenaeus chinensis by in situ hybridization

White Spot Syndrome Virus (WSSV) was purified from hemolymph of infected shrimp. After nucleic acid extraction from the purified virus particles, EcoR I-digested fragments of the WSSV genome were cloned; three of these fragments were used as non-radioactive probes labeled with DIG-11-dUTP. The probes hybridized in situ, with sections located in the nuclei of all WSSV-infected tissues. The virus was detected in the gill, stomach, epidermis, and connective tissue and so on, but not detected in healthy shrimp tissues and epithelial cells of hepatopancreatic tubules of diseased shrimp. This work was supported by SRF for ROCS, SEM and the National 863 Project (Grant 819-Q-08) and Project under major State Basic Research Development Program (Grant G1999012002).     

ON THE MARINE CILIATE, Pseudocohnilembus hargisi （PROTOZOA, CILIOPHORA, SCUTICOCILIATIDA） WITH COMPARISON OF ITS RELATED SPECIES

The living morphology, nuclear apparatus, infraciliature and silverline system of the poorly-known ciliatePseudocohnilembus hargisi Evans & Thompson, 1964, collected from a shrimp culture pond in Qingdao, China, were examined by observation of the live specimen; and by the pyridinated carbonate method and Chatton-Lwoff technique. The main characteristics: 13–14 (usually 13) longitudinal or slightly spirally arranged somatic kineties; single contractile vacuole with usually 2 (1–2) pores at posterior ends of the fourth and fifth somatic kineties to the right of the buccal cavity. Several related congeners were compared and morphometrically tabulated. Project No. 39970098 supported by NSFC.     

Effects of several immunostimulants on phenoloxidase and hemocytes of the crab Charybdis japonica

To investigate the stimulating effects of immunostimulants on the autogenous immunocompetence of crabs and the possible mechanisms involved, the immunostimulating effects of β-1,3-glucan, lipopolysaccharide (LPS), inactivated Vibrio harveyi and Vibrio anguillarum on phenoloxidase (PO) and hemocytes of Charybdis japonica were investigated in this study. It was found that the yields and the enzymatic activities of purified PO in C. japonica increased significantly after the crabs were treated with immunostimulants, while the unit enzymatic activities remained almost the same. After treatment with β-1,3-glucan and LPS, the amount of rough endoplasmic reticulum (RER) and the number of mitochondria in both semigranular cells and granular cells increased greatly, and the number of cytoplasmic granules decreased but with enlarged volume. However, the corresponding characteristics of hyaline cells remained almost the same. On the other hand, the number of granules in semigranular cells decreased greatly, and the number of mitochondria of hyaline cells increased greatly, after treatment with inactivated vibrios. It may be concluded that the effect of polysaccharide immunostimulants on the innate immune system of C. japonica is different from that of inactivated vibrio immunostimulants. The immunity-enhancing mechanism of polysaccharides in crab autogenous immunocompetence is probably accomplished by the increased yields of PO and total PO activities, while that of inactivated vibrios is probably accomplished by the partially increased yields of PO and total PO activities as well as the significantly improved phagocytotic abilities of semigranular cells and hyaline cells.     

Genetic improvement on Chinese shrimp （Fenneropenaeus chinensis）： growth and viability performance in F1 hybrids of different populations

Fenneropenaeus chinensis distributed in the Yellow Sea and Bohai Sea of China and the west coast of the Korean Peninsula. Different geographical populations represent potentially different genetic resources. To learn further the characteristics of different geographical population, crosses among two wild and three farmed populations were produced. The two wild populations were from the Yellow Sea and Bohai Sea （WYP）, and the west coast of the Korean Peninsula and coast （WKN）. The three farmed populations included the offspring of first generation of wild shrimp from coast in Korea （FKN）, the Huang Hai （the Yellow Sea in Chinese） No.1 （HH1）, and JK98. The phenotypes growth and survival rates of these populations were compared to confirm the feasibility for crossbreeding. The body length （BL）, carapace length （CL）. carapace width （CW）, height of the second and third abdominal segment （HST）, width of the second and third abdominal segment （WST）, length of the first abdominal segment （LF）, length of the last abdominal segment （LL）, live body weight （BW）, and survival rate were measured, Different combinations were statistically performed with ANOVA and Duncan＇s Multiple Range Test. The results show that the survival rate of JK98（♀）×WKN（♂） was the highest, followed by WYP（♀）×WKN（♂）, FKN（♀）×WYP（♂）, FKN（♀）×HHI（♂） and WYP（♀）×FKN（♂）; the body weight of FKN（♀）×HHI（♂） was the highest, followed by FKN（♀）×WYP（♂）, WYP（♀）×WKN（♂）, WYP（♀）×FKN（♂） and JK98（♀）×WKN（♂）; the total length had the same ranking as the body weight. All growth traits in hybrids JK98（♀）×WKN（♂） were the lowest among all combinations. F1 hybrids had significant difference （P〈0.05） in BL, CL, HST, LL, and BW; and insignificant difference （P〉0.05） in other growth traits and survival rate. The results of Duncan＇s Multiple Range Test are that BL and CL of J     

On a new species ofPavlova (Prymnesiophyceae) from China

Pavlova viridis sp. nov. is described on the basis of light and electron microscope observations. The material was collected from the coast of haiyang county of Shandong, China. It is characterized by 1) yellowish green to green chloroplast, 2) rudimentary short flagellum 0.3 μm long and hook-shaped, 3) long flagellum with small spherical knob-scales, 4) absence of pyrenoid and stigma, and 5) marine habitat. This new species andP. salina seem to be most closely related to each other but their colour, the insertion of their three appendages, the shape of their knob scales, and their cell periplasts are quite different from one another. Contribution No. 1879 from the Institute of Oceanology, Academia Sinica     

A NEW SPECIES OF RANINOIDES （CRUSTACEA ： DECAPODA ： RANINIDAE） FROM OFF THE SOUTHEASTERN COAST OF HAINAN ISLAND AND NANSHA ISLANDS

Ranlnoides longifions new species, off SE coast of Hainan Island, dorsal surface of carapace with an arched granular edge across carapace between the anterolateral teeth, median frontal tooth longest, second frontal teeth subquadrate.