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 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.     

STUDIES ON IMMUNODIAGNOSIS OFHEPATOPANCREATIC PARVO- LIKEVIRUS DISEASE OF THE CHINESEPENAEID, PENAEUS CHINENSISI. PURIFICATION OF THE VIRUS

The viral disease of penaeids is indistinct 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 cultured Penaeus chinensis was conducted using immuno-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 chinen- sis 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.     

SELENIUM REQUIREMENT OF SHRIMP PENAEUS CHINENSIS

Penaeus chinensis were reared in fibreglass tanks for the study of their selenium requirements. The shrimp were fed semipurified diets containing graded levels of selenium, and weight gains, activities of glutathione peroxidase (GSH-Px) and selenium contents in muscle and hepatopancreas were determined. Weight gain and GSH-Px activity were the highest when the shrimp were fed diet containing 20 mg/kg selenium; Good linear correlation was found between GSH-Px activities and selenium contents in the diets, and the number of healthy shrimp. The experiment showed that 20 mg/kg selenium in the diet is optimal for the shrimp and that GSH-Px activity can be an important biochemical index of the selenium nutrition status of the animal.     

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 fishmeal, peanut cake, corn meal, soybean cake, wheat bran, vitamin mix and mineral mix, and supple-mentations of potassium, sodium, magnesium and iodine in the basal diet were made according to theL_9(3~4) orthogonal table. The results indicatal that iodine supplementation improved growth of the shrimpsignificantly and raised survival very significanly, iodine requirement of the shrimp was 0.003%, sodiumrequirement was 0.87% or less; and that 1.1-1.3% potassium and 0.18-0.38% magnesium in the dietwere proper nutrition supplements for the shrimp.     

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(100g)-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 complement3-1ike and complement4-1ike 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.     

斑节对虾白斑综合症杆状病毒感染的实验模式

在建立斑节对虾实验室养殖模式的基础上，对野外采集的有白斑综合症病毒（WSSV）病典型症状的斑节对虾初步纯化其病毒，并进行WSSV的PCR检测，得到WSSV的感染用样品。对健康斑节对虾分别进行浸浴感染，投喂感染和注射感染。对感染死亡个体进行WSSA的PCR检测和细菌检测，证实WSSV感染性和致死性。浸浴感染、投喂感染和注射感染的感染量分别为4mL/L、0.2g/10g虾体、1/2稀释液0.05mL/10g虾体，死亡开始时间分别为16d、42h、28h，三种感染模式最终死亡率100%，从开始死亡到全部死亡延续时间分别为15d、82h、44h。     

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.     

POPULATION DYNAMICS AND PRODUCTION OF NEANTHES JAPONIC A (IZUKA)IN A SHRIMP POND

During late February to mid-March, 1991, when mature N. japonica swarmed in the sea surface off Jimo County, Qingdao, seawater containing numerous early embryos was pumped into a nearby 2.7 ha. muddy-sand shrimp pond, where the embryos continued to develop. 800000 Penaeus chinensis seedlings were introduced into the pond on May 30, 1991. Monthly benthic samplings were carried out to determine the population dynamics and production of N. japonica in this atypical environ -ment.The density of N. japonica varied from 0 to 20400 ind./m2. The predation of P. chinensis was considered to be responsible for the mass mortality of N. japonica in June through July, when the worms were 2.2-3.3cm in length and lived in the top 2-4cm sediment. From August to September of 1991, the hottest period in the site, there was no substantial mortality of N. japonica as reported in earlier literature. This was explained by the deeper burrowing of the' worms that protected them from predation by shrimps.Monthly biomass ranged f     

对虾病毒病暴发前期病毒和弧菌相互作用关系

模拟天然继发感染研究对虾病毒病暴发前期病毒和弧菌的关系。结果显示 ,弧菌先感染再病毒感染组比病毒先感染再弧菌感染组死亡率高、死亡快、生长慢、免疫功能低下。表明弧菌的潜伏感染对病毒的增殖有利 ,而病毒的潜伏感染对弧菌的继发感染没有明显的促进作用。说明病毒性流行病的暴发前期可能是由弧菌先感染 ,使对虾体内环境发生变化而助长病毒感染 ,其中对虾的免疫力下降是关键     

生物生态防治虾病研究

本文探索虾病防治新途径,采用低值海藻江蓠和对虾混养的生态学方法,江蓠吸收水中二氧化碳进行光合作用,产生大量氧气;同时吸收有机物的分解物 NH_3—N,加速有机物分解。减少 H_2S 积累;从而消除水中紧迫因子威胁,大大减少池水自污及病害,起到抑菌、抑虫、净化水质作用,提高成活率,试验圹对虾成活率达95.1%,对照圹56.6%。试验结果表明,虾病防治效果显著,经济效益明显。     

Hematological Changes m 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 manifesta- tions 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 dis- integrated 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 bemocytes 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.     

湛江港区底泥悬浮物对凡纳滨对虾仔虾成活的影响

模拟吹填、疏浚等海上施工过程所产生的底泥悬浮物浓度的变化规律,探讨不同底泥悬浮物浓度下对凡纳滨对虾仔虾成活影响。结果表明:海水中底泥悬浮物浓度超过10mg/L,持续时间超过12h时,将对凡纳滨对虾成活率产生影响,随着悬浮物实验浓度的递加,仔虾的死亡率从13.33%增至96.67%;超过80mg/L,且持续时间超过48h,将对凡纳滨对虾产生严重影响,甚至导致全部仔虾的死亡;悬浮物对凡纳滨对虾的12h、24h、36h和48h半致死量(LC50)分别为54.56、40.29、25.8和19.18mg/L。     

抗菌肽对凡纳滨对虾生长和机体免疫的影响

基础饲料(对照组)中添加10g/kg抗菌肽制剂喂养体重0.10±0.01g、体长2.17±0.11cm的凡纳滨对虾(Penaeus vannamei)30d,进行10d的水浴攻毒(鳗弧菌10~8/mL),探讨饲料中添加抗菌肽对凡纳滨虾生长、成活率、免疫保护率,以及肌肉中溶菌酶活性、超氧化物歧化酶活性、总抗氧化能力的影响。结果表明:饲料中添加抗菌肽可显著提高凡纳滨对虾相对增重率、成活率和对致病菌的免疫保护率(P<0.05),但对饲料系数无显著影响(P>0.05);未受外界微生物刺激时,饲料中添加抗菌肽对凡纳滨对虾总抗氧化能力有促进作用(P<0.05),受外界微生物刺激后,饲料中添加抗菌肽喂养凡纳滨对虾可提高对虾超氧化物歧化酶活性(P<0.05)和总抗氧化能力(P<0.01)。因此饲料中添加抗菌肽喂养凡纳滨对虾可提高其机体非特异性免疫。     

4种斑节对虾亲虾饵料蛋白质的营养价值评价

分析了沙蚕、缢蛏、拖鱿鱼胴体、花蟹蟹肉以及斑节对虾亲虾肌肉和成熟卵巢基本营养成分、氨基酸组成。通过计算4种饵料与斑节对虾亲虾肌肉和成熟卵巢的必需氨基酸比率(A/E)的比值和必需氨基酸指数(EAAI),评价4种斑节对虾亲虾饵料蛋白质的营养价值。结果表明:沙蚕、缢蛏、拖鱿鱼胴体、花蟹蟹肉以及斑节对虾(Penaeusmonodon)亲虾肌肉和成熟卵巢中的粗蛋白质量分数分别为9.18%、8.14%、12.10%、17.00%、21.90%、18.60%;总氨基酸质量分数分别为7.30%、7.47%、11.70%、14.80%、19.40%、14.60%。以斑节对虾亲虾肌肉蛋白为参比,沙蚕、缢蛏、拖鱿鱼胴体以及花蟹蟹肉的必需氨基酸指数分别0.93、0.94、0.94、0.97;以斑节对虾亲虾成熟卵巢蛋白为参比,沙蚕、缢蛏、拖鱿鱼胴体以及花蟹蟹肉的必需氨基酸指数分别0.95、0.93、0.88、0.84。这说明4种亲虾饵料相对斑节对虾亲虾而言是一种优质蛋白质,能够满足亲虾蛋白需求。不过,精氨酸、组氨酸等必需氨基酸,不能完全满足亲虾需求。     

恩诺沙星在凡纳滨对虾体内的代谢和残留消除规律

在26±2℃水温下,每天投喂含有恩诺沙星药物的饲料,研究恩诺沙星在凡纳滨对虾Litopenaeus vanname肌肉、肠和肝胰脏组织中的代谢和残留消除规律。残留药物用乙腈提取,液相色谱串联质谱仪检测。结果表明:恩诺沙星在凡纳滨对虾体内可代谢为环丙沙星,对虾体内同时有恩诺沙星和环丙沙星两种药物残留;环丙沙星在肌肉、肝胰脏和肠组织中的消除时间分别为6、8、10 d,而恩诺沙星在这三组织中的消除时间则为12、14、16 d。建议把肠作为该药残留监控的靶组织,凡纳滨对虾的休药期不少于16 d。     

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.     

凡纳滨对虾和斑节对虾对WSSV敏感性的比较

用6.0×104拷贝、1.2×104拷贝和6.0×103拷贝3种剂量白斑综合症病毒(WSSV)对凡纳滨对虾和斑节对虾进行人工注射感染,比较了两种对虾对WSSV敏感性的差异。结果表明,凡纳滨对虾死亡时间随病毒剂量降低而延长,斑节对虾死亡时间没有明显差异;随病毒剂量的降低,凡纳滨对虾人工注射感染后病毒复制高峰时间显著延长,斑节对虾感染后病毒复制高峰时间相同,WSSV在凡纳滨对虾体内比在斑节对虾体内复制慢。对虾携带WSSV数量最低为3.3×107拷贝.g-1,最高为4.3×108拷贝.g-1。凡纳滨对虾比斑节对虾对WSSV的抵抗性更强。