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 of Penaeus chinensis (EEBV-PC, de-tected by the authors in 1993), infections hypodermal and hematopoietic necrosis virus, andhepatopancreatic parvo-like virus, and that the first two viruses seem to be the main pathogens of theepidemic in the northern regions in 1993.     

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.     

The Complete Sequence of Mitochondrial COⅡ Gene of Fenneropenaeus chinensis and Its Applicability as a Marker for Phylogenetic Analysis

The complete mitochondrial cytochrome oxidase subunit Ⅱ （COⅡ） gene of Penaeinae shrimp Fenneropenaeus chinensis was cloned and sequenced. The gene is 688 bp in length and codes for 229 amino acids. It shows 83.2%, 87.0% and 83.8% sequence similarity to Marsupenaeus Japonicus, Penaeus monodon and Farfantepenaeus notialis, respectively. The A＋T content of the whole gene and that at the third position of codons are 64.7% and 78.2%, respectively. The phylogenetic relationship between F. chinensis and three other species representing genera Farfanatepenaeus, Marsupenaeus and Penaeus was analyzed. Results showed that the genetic distances among the four taxa ranged from 0.144 0 to 0.200 5, exceeding those estimated with COⅠ and partial 16S rRNA gene sequences among Marsupenaeus, Litopenaeus and Melicertus, and being therefore larger than the value among subgenera. It has been suggested that the COⅡ gene has a faster evolutionary rate than that of the COⅠ gene and partial 16S rRNA gene and could be used for phylogenetic analysis at genus or species level. The results of the present study indicated that Farfantepenaeus, Fenneropenaeus, Marsupenaeus and Penaeus are at a higher phylogenetic level than subgenus, which supports the opinion of the elevation of phylogenetic status of the four subgenera to genus level.     

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.     

Triploidy induction with heat shocks toPenaeus chinensis and their effects on gonad development

Heat shocks effectively produced triploids inPenaeus chinensis. Fertilized eggs heat shocked (28–32°C) for 8 to 16 minutes, starting from 8 to 20 minutes after spawning, resulted in triploidy induction rates of 39%–75%. Several triploid populations were cultured to 10 cm. In a triploid population, two kinds of ovaries were observed. Histological examination showed apparent differences between these two kinds of ovaries, whereas among male shrimp, there were no such differences. Contribution No. 3465 from the Institute of Oceanology. Chinese Academy of Sciences. The study was supported by the International Foundation for Science, Asian Fisheries Society and Chinese Climbing Project PDB 6–2, and Chinese National 863 Project (No. 863-819-01-05).     

Research on heavy metals in Ruditapes philippinarum and soda industry wastes

Heavy metals pollution in Jiaozhou Bay was studied inRuditapes. philippinarum, a bioindicator of seawater pollution. Heavy metals in soda industry wastes “white mud” were also studied. Comparison of microwave digestion method with general digestion method revealed that microwave digestion is superior to general digestion in precision, recovery, digestion speed and efficiency, etc. Cd concentration in HongdaoR. philippinarum samples exceeded the national standard by 0.046 mg/kg, that of Yinghai sample by 0.02 mg/kg, and that of Hongshiya sample by 0.22 mg/kg. Sample Pb concentration in Hongshiya was found to exceed the national standard by 0.02 mg/kg. However the heavy metals concentration inR. philippinarum near the Qingdao Alkaline Factory was complied with the standard. This was proved byPenaeus chinensis culture experiment. Therefore, the possible contamination source may come from other land areas. This study is supported by Project 973 (NO.2002CB 412402), and key project of the Chinese Academy of Sciences (Grant NO.KJCX315W-215).     

Comparative studies on the larval development of the penaeid shrimps,Penaeus Chinensis,P. merguiensis andP. penicillatus

The morphology of larval and 1st postlarval stages ofPenaeus penicillatus are described. Results from comparative studies on larval development ofP. penicillatus, P. merguiensis andP. chinensis are as follows: These three species could not be identified during their naupliar stages. In the 1st protozoea, the antennule L1/L2 value is 1.7–2.0 inP. merguiensis, but less than 1.7 inP. chinensis andP. penicillatus; in the 2nd protozoea, the supra-orbital spine inP. chinensis is not bifurcated, while those ofP. merguiensis andP. penicillatus are bifurcated; in the 3rd protozoea, there is a minute (or no) dorso-median spine on the posterior margin of the 1st and 2nd abdominal somite inP. chinensis, but they are prominent inP. merguiensis andP. penicillatus. In the mysis and 1st postlarval stages,P. chinensis differs fromP. merguiensis andP. penicillatus in having 9 (8 in the other 2 species) long setae on the exopod of pereopods 1–3; additionally, one dorsal tooth appears on the rostrum ofP. chinensis in the 2nd mysis and that of the other 2 species in the 3rd mysis;P. chinensis has 2 (mostly) or 1 dorsal tooth on the rostrum in the 3rd mysis and 2–3 in the 1st postlarva, whileP. penicillatus andP. merguiensis have only 1 in the 3rd mysis and 1st postlarva. Comparative studies on larval development showedP. penicillatus has closer affinity withP. merguiensis than withP. chinensis. Contribution No. 2218 from the Institute of Oceanology, Chinese Academy of Sciences     

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.     

Effects of temperature and dissolved oxygen content on oxygen consumption rate of Chinese prawn, giant tiger prawn and giant freshwater prawn

INTRODUCnON-WatertemperatheanddissolvedoxygenareimportantenvironmentalfactorsinthegmwthanddevelOPmntofaqUaticorgAnsms.ThereareInanPublishedstUdiesontheox-.ygenconsUInPtionrate(OCR)oftheprawn')(WangJim,l986;WangDanghenetal-,199l;ZangWeilingetal-,l992,l993),buttheydidnotcontalndetailedinfonnaionabouttheeffectoftemPerabeontheOCR-ThePresentstudydetendnedtheOCRofju-venilesofChineseprawn,gianttiserprawnandgiantfreshwaterprawnthatwererearedatdifferenttemperatllresforaboutaweekattheCaoj…     

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.     

Chromosomal localization of 5S rDNA in Chinese shrimp (Fenneropenaeus chinensis): a chromosome-specific marker for chromosome identification

Chinese shrimp (Fenneropenaeus chinensis) is an economically important aquaculture species in China. However, cytogenetic and genomic data is limited in the organism partly because the chromosomes are difficult to isolate and analyze. In this study, fluorescence in-situ hybridization (FISH) was used to identify the chromosomes of F. chinensis. The 5S ribosomal RNA gene (rDNA) of F. chinensis was isolated, cloned and then used as a hybridization probe. The results show that the 5S rDNA was located on one pair of homologous chromosomes in F. chinensis. In addition, triploid shrimp were used to evaluate the feasibility of chromosome identification using FISH and to validate the method. It was confirmed that 5S rDNA can be used as a chromosome-specific probe for chromosome identification in F. chinensis. The successful application of FISH in F. chinensis shows that chromosome-specific probes can be developed and this finding will facilitate further research on the chromosomes of penaeid shrimps.     

Requirements of shrimp,Penaeus chinensis O'sbeck for potassium,sodium, magnesium and iodine

Potassium, sodium, magnesium and iodine requirements of shrimp,Penaeus chinensis were studied. Orthogonal design was employed in this experiment. The composition of the basal diet consisted of fish meal, peanut cake, corn meal, soybean cake, wheat bran, vitamin mix and mineral mix, and supplementations of potassium, sodium, magnesium and iodine in the basal diet were made according to the L₉(3⁴) orthogonal table. The results indicated that iodine supplementation improved growth of the shrimp significantly and raised survival very significantly, iodine requirement of the shrimp was 0.003%; sodium requirement was 0.87% or less; and that 1.1–1.3% potassium and 0.18–0.38% magnesium in the diet were proper nutrition supplements for the shrimp. Contribution No. 2342 from the Institute of Oceanology, Chinese Academy of Sciences.     

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.     

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.     

Partial sequence analysis of mitochondrial COI gene of the chinese shrimp, <Emphasis Type="Italic">fenneropenaeus chinensis</Emphasis>

Eight hundred and thirty eight base pair fragment of mitochondrial COI gene of wild and cultured populations (CP1, CP4, CP5 and CP6) of Fenneropenaeus chinensis was amplified and sequenced. The A, T, G and C contents of the sequence were 235 bp (28.0%), 307 bp (36.6%), 138 bp (16.5%) and 158 bp (18.9%), respectively. Furthermore, 556 bp fragment of the sequence was used to discuss the phylogenetic relationship among 14 Penaeidae species using Alpheus armillatus as the outgroup. From the molecular phylogenetic tree constructed by neighbor-joining method, we obtained three large shrimp groups: Farfantepenaeus, Litopenaeus and Fenneropenaeus group. The results also indicated that there were a closer genetic relationships between F. aztecus and F. paulensis, L. schmitti and L. setiferus, F. indicus and F. merguiensis, and the genus Farfantepenaeus was closer to Litopenaeus.     

Morphological Comparison between Chinese Ayu and Japanese Ayu and Establishment of Plecoglossus altivelis chinensis Wu ＆ Shan subsp, nov.

The countable characters of Qingdao, Zhejiang, Liaoning and Japanese Ayu are compared. There are no substantial different characters among the former three Ayu, so the comprehensive values of them are taken as representative of Chinese Ayu, which is in turn compared with Japanese Ayu (Plecoglossus altivelis altivelis and Plecoglossus altivelis ryukyuensis). By the coefficient of difference test, 3 and 4 characters are found to be beyond the subspecies level between Chinese Ayu and nominate subspecies (Plecoglossus altivelis altivelis) and between Chinese Ayu (Plecoglossus altivelis chinensis) and Ryukyus subspecies (Plecoglossus altivelis ryukyuensis), respectively, which shows that they belong to different subspecies. The China mainland Ayu is a new subspecies, Plecoglossus altivelis chinensis Wu & Shan, the establishment of which and its forming causes are discussed.     

Feeding habits and food composition of half-fin anchovy,Setipinna taty (C et V) in the Bohai Sea

Analysis of the contents of 3,479 stomachs of half-fin anchovy (taken monthly from April 1982 to May 1983 from the Bohai Sea) shows that half-fin anchovy is a zooplanktivorous fish; the young fish feeds mainly on Copepoda while the adult fish feeds mainly on Mysidacea andAcetes chinensis; the food composition has a seasonal change, the main food is Mysidacea in spring butAcetes chinensis in summer and fall; the feeding intensity is closely related to its spawning. Contribution No. 1781 from the Institute of Oceanology, Academia Sinica.     

Screening white spot syndrome virus (WSSV)-resistant molecular markers from <Emphasis Type="Italic">Fenneropenaeus chinensis</Emphasis>

White spot syndrome virus (WSSV)-resistant molecular markers were screened from the selectively bred new variety ‘Huanghai No. 2’ of Fenneropenaeus chinensis using unlabeled-probe high-resolution melting (HRM) technique. After the artificial infection with WSSV, the first 96 dead shrimps and the last 96 surviving shrimps were collected, representing WSSV-susceptible and -resistant populations, respectively. The genotypes at well-developed 39 single nucleotide polymorphisms (SNPs) loci were obtained. As revealed in the Chi-square test, 3 SNPs, genotype A/A of contig C364-89AT, genotype A/A of C2635-527CA and genotype C/T of contig C12355-592CT, were positively correlated with disease-resistance traits. Other 2 SNPs, genotype G/G of contig C283-145AG and genotype C/C of contig C12355-592CT, were negatively correlated. Moreover, analysis with BlastX program for disease-resistant SNPs indicated that 3 contigs, Contig283, Contig364 and Contig12355, matched to the functional genes of effector caspase of Penaeus monodon, peptide transporter family 1-like protein, and 40S ribosomal protein S2 of Perca flavescens with high sequence similarity. The results will be helpful to provide theoretical and technical supports for molecular marker-assisted selective breeding of F. chinensis.     

Purification and characterization of an alkaline protease from Acetes chinensis

An alkaline protease from Acetes chinensis was purified and characterized in this study. The steps of purification include ammonium sulfate precipitation, ion-exchange chromatography with Q-sepharose Fast Flow, gel filtration chromatography with S300 and the second ion-exchange chromatography with Q-sepharose Fast Flow. The protease was isolated and purified, which was present and active on protein substrates (azocasein and casein). The specific protease activity was 17.15 folds and the recovery was 4.67. The molecular weight of the protease was estimated at 23.2 kD by SDS-PAGE. With azocasein as the susbstrate, the optimal temperature was 55°C and the optimal pH value was 5.5. Ion Ca²⁺ could enhance the proteolytic activity of the protease, while Cu²⁺, EDTA and PMSF could inhibit its activity.