Detection of Penaeus monodon-type baculovirus (MBV) infection in Penaeus monodon Fabricius by in situ hybridization

Abstract. A digoxigenin-labelled DNA probe was used for in situ detection of the Penaeus monodon -type baculovirus (MBV) derived from cloned MBV polyhedrin genome in cultured Penaeus monodon Fabricius. First, the specificity of the probe against MBV DNA with dot blot hybridization analysis was verified. This probe indicated that cloned MBV polyhedrin fragment can be used as an MBV-specific probe. This was then used to microscopically examine sections of MBV-infected tissues for a blue-purple precipitate indicative of a positive reaction for MBV. MBV-positive cells were located only in the epithelium of the hepatopancreatic tubules and of the midgut. Furthermore, comparison of the susceptibility to MBV infection among several life-stages of the shrimp showed that the MBV genome was found in the zoea, mysis, post-larva, and adult stages, whereas MBV DNA was not detected in either eggs or nauplii. The results were quantified from in situ hybridization with an image analyser to compare the degree of cell infection among groups of cultured P. monodon collected from various farms in Taiwan.     

斑节对虾杆状病毒在感染对虾中肠腺中的分布

利用组织切片法研究了斑节对虾杆状病毒（MBV）在养殖早期班节对虾及养成期斑节对虾中肠腺各部位的分布特点。斑节对虾的中肠腺可分为头部、亚头部、中部、亚尾部和尾部等5个部分，其上皮细胞包括E细胞、F细胞、B细胞和R细胞等4种类型。头部和尾部均以E细胞为主、亚头部、中部和亚尾部均以R细胞为主。在感染度较轻的样本中，MBV一般只感染F细胞、R细胞和B细胞。感染度较重时，E细胞也同时受累。在养殖早期虾和养成期虾中，MBV相对感染度在中肠腺各部分的分布均以中部最高，而且以中部的中间最高，离中部越远，相对感染度越低。     

Development of a Penaeus monodon-type baculovirus (MBV) DNA probe by polymerase chain reaction and sequence analysis

Abstract. Penaeus monodon -type baculovirus (MBV) was isolated and purified from the hepatopancreases of MBV-infected Penaeus monodon Fabricius. MBV DNA was extracted and used as a template in a polymerase chain reaction (PCR). The primers were chosen from conserved regions of the polyhedrin gene of Autographa californica nuclear polyhedrosis virus (AcNPV). One DNA fragment (674 base pairs) was amplified after PCR. There was a 65% homology between the predicted amino acid sequence of this PCR product with that of the polyhedrin polypeptide of AcNPV. Nucleotide sequence analysis indicated that the amplified DNA is the open reading frame of the MBV polyhedrin gene. This 674 bp DNA fragment was subsequently used as a probe in a dot blot analysis. The probe was able to hybridize with the DNA extracted from the purified MBV and from the MBV-infected P. monodon , but not from the MBV uninfected P. monodon.     

蛋白水平对不同斑节对虾家系生长与消化酶活性的影响

为探究饲料蛋白水平与不同斑节对虾(Penaeus monodon)家系生长和消化酶之间的关系,研究了3种蛋白水平(34%、38%、42%)的饲料对16个斑节对虾家系的生长性状以及胃和肝脏消化酶活性的影响。经过56 d的养殖实验,结果表明:(1)随着蛋白水平的升高,所有斑节对虾家系的特定生长率均有所增加,F1、F2、F4、F7、F12、F13、F15等7个家系的特定生长率在34%、38%、42%3种蛋白水平组未见显著性差异(P>0.05),其余家系在3种或者其中的2种饲料之间有显著性差异(P<0.05);(2)斑节对虾特定生长率在家系和饲料蛋白质水平这两个因素间交互作用不显著(P=0.974);(3)所有斑节对虾家系的蛋白酶活性均随着饲料蛋白含量的增加而增加,淀粉酶活性有所下降,但规律不明显;(4)13个斑节对虾家系的肝脏蛋白酶活性、14个家系的胃蛋白酶活性均与家系特定生长率有着极显著的相关性(P<0.01),淀粉酶活性和特定生长率的相关性远小于蛋白酶活性。     

冰冻斑节对虾体内病毒侵染中国对虾的电镜观察

用市售冰冻斑节对虾中头胸甲有白斑的病虾做感染源，对中国对虾仔虾进行人工感染，通过电子显微技术，对中国对虾仔虾的肝胰腺细胞进行观察研究。结果表明；冰冻斑节对虾体内的杆状病毒能够使中国对虾致病，其发病程度与对虾养殖密度有关。从病毒基质发生到形成完整的病毒粒子及侵染大致分4个阶段；病毒基质发生期，病毒囊膜形成期，病毒粒子装配期，病毒粒子再度侵染期。同时观察中还发现了一些可能与病毒形成有关的特殊物质，如绒团物和冰晶状物等。     

Recent Advances in Nutrition of Penaeus monodon

This paper provides a brief summary of information on recent advances in the nutrition of Penaeus monodon . The nutritional requirements of P. monodon have been little known despite its importance in aquaculture. Recent findings indicate a noticeable difference between some important nutrient requirements of P. monodon and P. japonicus , the most studied shrimp species. For example, the thiamin requirement for P. monodon was estimated to be 14 mg/kg diet, while for P. japonicus it is 60-120 mg/kg diet for juveniles and 40-80 mg/kg diet for larvae. The same contrast was also observed in the requirements of riboflavin (22.5 mg/kg in juvenile P. monodon vs. 80 mg/kg in larval P. japonicus ), niacin (6.4 mg/kg vs. 400 mg/kg) and vitamin C (2,000-2,500 mg/kg vs. 10,000 mg/kg).
Research efforts to understand the lack of ability for shrimp to effectively use dietary crystalline amino acids have resulted in the development of a microencapsulated amino acid that can be assimilated by P. monodon which is incapable of utilizing the essential amino acid in crystalline form. This development enables the quantification of requirements of the amino acid and other essential amino acids for shrimp. A study using cellulose-acetate-phthalate encapsulated arginine indicated that the arginine requirement of postlarval P. monodon is 5.47 g/100 g protein (2.50 g/100 g diet).
Other aspects concerning the importance of digestive enzymes, dietary requirements of protein, carbohydrate, lipid and vitamin, energy and larval feeding for P. monodon were reviewed.     

Evidence for the presence of white spot syndrome virus (WSSV) and monodon baculovirus (MBV) in wild Penaeus monodon (Fabricius) broodstock,in the southeast coast of India

A survey on the presence of the viruses of two economically significant diseases, white spot syndrome virus (WSSV) and monodon baculovirus (MBV) in wild‐collected Penaeus monodon broodstock, was conducted during different seasons of the year in two major coastal areas of southeast India. The broodstock were collected along the coast of Tamil Nadu and Andhra Pradesh during summer, premonsoon, monsoon and post‐monsoon seasons for three consecutive years. A total of 7905 samples were collected and subjected to MBV screening, and 6709 samples that were screened as MBV negative were diagnosed for WSSV. MBV was detected using rapid malachite green staining and WSSV by nested polymerase chain reaction. Prevalence data of the viruses were analysed using the EpiCalc 2000 program at 95% confidence interval. Samples collected from the Andhra Pradesh coast displayed a slightly higher prevalence of WSSV and MBV infection than those collected from Tamil Nadu, although this difference was not statistically significant (P > 005). In addition, it was found that the prevalence of both WSSV and MBV infections fluctuated according to season. Data on prevalence of these viruses in broodstock would be useful to develop strategies for shrimp health management along the southeast coast of India.     

上海地区低盐海水中斑节对虾养殖试验

本试验共进行了两年。除88—2池跑苗外,试验组成活率达30%,产量均达到150公斤/亩左右,其中88—1池最高,为226.8公斤/亩。斑节对虾日生长率超过0.90毫米,与对照的中国对虾比,成活率、生长速度均高于对照组。在密度相近的条件下,生长速度快于中国对虾20%以上;饵料系数低于中国对虾,产量可达到或超过中国对虾产量,而生长期却缩短20天左右。试验结果说明,斑节对虾在上海地区养殖是完全可行的。     

池塘养殖斑节对虾生长、发育与性成熟

为了解不同养殖条件下斑节对虾的生长、外生殖器发育、性腺发育及性成熟之间的关系,对其养殖进行跟踪调查研究.结果显示:①斑节对虾雌雄外生殖器官发育和头胸甲长呈线性关系;②不同养殖环境条件下,斑节对虾性成熟生物学最小型个体无显著差异.雄性精荚出现的生物学最小型个体为头胸甲长3.1 cm,体长11.1 cm,体质量20.0 g;雄性性成熟个体的头胸甲长3.7 cm,体长13.0 cm,体质量37.0 g.池养雌性斑节对虾的性成熟生物学最小型个体以纳精囊的发育完全(可与雄虾交配)为标志,其最小性成熟个体的头胸甲长4.3 cm、体长15.1 cm、体质量53.0 g,雌性性成熟个体为头胸甲长5.0 cm,体长17.0 cm,体质量75.0 g以上;③池塘养殖斑节对虾性成熟与日龄和养殖环境相关.鱼塭雄虾精荚出现的最早时间为日龄120 d前后,其性成熟日龄约为160 d;池塘养殖雄虾精荚出现的最早时间为日龄150 d前后,其性成熟日龄约为260 d.鱼媪雌虾最早交配发生在日龄165 d前后,性成熟日龄205～236 d,池养雌虾最早交配发生在日龄240～ 280 d,性成熟日龄295～360 d以上.     

A histopathological disease survey of cultured shrimp in North East Sumatera, Indonesia

Abstract. A disease survey was conducted in North East Sumatera, Indonesia, during June and July 1991. The objeet was to obtain information regarding the diseases present in shrimp culture faeilities. Shrimp were sampled for histopathological examination and details of the farm management and environment were recorded. The stocks sampled included some reported to be suffering from disease problems and some reported to be in good health. Samples were taken from 24 pond farms, five hatcheries and a broodstock fisherman. All farms and hatcheries cultured Penaeus monodon Fabricius, one farm also cultured Penaeus merguiensis de Mann, and two cultured Metapenaeus spp. Several wild caught P. monodon female broodstoek and some wild rice shrimp ( Acetes spp.) were also sampled. The disease conditions detected in P. monodon were monodon baculovirus (MBV), hepatopancreatic parvo-likc virus (HPV), septic hcpatopancreatic necrosis (SHN), bacterial septicaemia (BS), haernocytie enteritis (HE), lymphoid organ pathology (LOP), external fouling organisms (EFO), and a single unconfirmed case of infectious hypodcrmal and haemopoietic necrosis (IHHN). The Acetes spp. from one site had a microsporidean infection in the striated muscle of the abdomen. EFO was the most common disease condition, and was identified in 16 farms and three hatcheries. MBV was detected in 14 farms, three hatcheries and in one broodstoek. SHN was found in 13 farms with one case of concurrent BS and LOP was found in eight farms. HE was found in four farms and HPV was identified in shrimp with MBV on two farms and as the only disease in one broodstoek. The significance and implications of these findings for the developing industry in Indonesia arc discussed.     

斑节对虾虾苗活力检测方法

该研究通过肉眼观察、镜检,进行干露、饥饿、盐度突降、福尔马林等抗性试验,并采用病毒检测等方法,以期建立评估斑节对虾（Penaeus monodon）虾苗活力和质量标准。结果表明,斑节对虾健康虾苗具有趋光性、集群性,体表光洁,肌肉透亮,肠胃食物充盈等特性。测试虾苗干露时间以15min为宜,健康虾苗干露后能立即恢复活力,而病弱虾苗多出现死亡、昏迷现象;虾苗的成活率随饥饿时间的延长而降低,随福尔马林浓度升高和时间延长而降低,随盐度突降幅度增加而降低。健康虾苗能忍受100～200μL·L^-1福尔马林溶液30min,成活率近100％;在盐度20～30下虾苗的成活情况较好,而其在淡水中仅能存活1h。对虾苗进行病毒检测,可以避免养殖中因虾苗携带病毒而可能导致的病毒性疾病的暴发。     

提高斑节对虾人工育苗幼体成活率试验

1998年闽南地区斑节对虾人工育苗幼体成活率普遍很低。作者结合多年的斑节对虾育苗生产经验,采取综合的技术措施,进行提高幼体成活率的试验。从无节幼体到仔虾幼体后期(P_(10)—P_(16)),试验的总成活率为42.1％。     

斑节对虾养殖群体遗传多样性的同工酶和RAPD分析

采用聚丙烯酰胺凝胶电泳（PAGE）和RAPD方法对厦门养殖斑节对虾（Penaeus monodon Fabricius）群体的遗传多样性进行分析。9种同工酶共检测到21个座位，其中多态座位13个，多态座位比例为61．90％，预期杂合度0．151，观察杂合度0．120，Hardy—Weinberg遗传偏离指数（d）为-0．208，存在杂合子缺失。经x^2拟合度检验，多数座位偏离Hardy—Weinberg平衡，表明群体未达到随机交配。14个10bp引物共获得了83个标记，单个引物获得的标记数为2～11个，平均每个引物扩增出5．93个座位，其中多态标记数68个，多态位点比例为81．93％，杂合度为0．246，基因多样性为0．260，Shannon’S信息指数为0．397。两种方法均表明该斑节对虾养殖群体的遗传多样性水平较高，但可能已有近交衰退发生。     

Triacylglycerol Digestion by the Leader Prawn Penaeus monodon

Abstract.— Midgut gland extracts from leader prawn Penaeus monodon were evaluated for lipase activity and positional specificity using triolein as a substrate. Lipase activity of aqueous and acetone midgut gland extracts was screened using a mammalian serum lipase test kit and triolein/agar emulsion clearing. Using the clearing method lipase activity was identified in both aqueous and acetone extracts. However, no lipase activity was detected using the serum lipase test kit. In vitro hydrolysis of triolein by midgut gland lipase at pH 8.0 produced 1,2-diacylglycerol and free fatty acid as the sole digestion products indicating specificity for the 1-position of triacylglycerol. The specifcity of lipases from Penaeus monodon for the primary position of triacylglycerol is similar to that of manmalian pancreatic lipase and lipases from Homarus americanus and most fish.     

Phototrophic Bacteria as Feed Supplement for Rearing Penaeus monodon Larvae

Larvae of the giant tiger prawn Penaeus monodon were reared from the naupliar to pos-tlarval stage, based on a diet of waste-grown phototrophic bacterium Rhodovulum sulfido-philum , the marine diatom Skeletonema costatum , and brine shrimp Artemia The experimental design consisted of feeding the protozoea with diatoms and varying proportions of bacterial biomass as diet D0 (0%), D1 (1%), D2 (2%), D3 (3%), and D5 (5%), while mysis and postlarvae were fed the above diet supplemented with Artemia nauplii. Controls were not fed with bacteria biomass (diet D0). The addition of R. sulfidophilum biomass in the diet had significant effects on the growth and survival of P. monodon larvae. In the first experiment, the larvae fed with diet D1 ( Skeletonema + 1% R. sulfidophilum ) reached the first postlarval stage (PL1) on the ninth day of feeding and had a mean total length (TL) of 6.13 ± 0.05 mm, which was significantly higher ( P < 0.001) than TL of larvae fed with the other three diets. In another experiment, prawn larvae fed with diet D2 had the highest overall survival of 46% and a significantly larger mean TL of 6.88 ± 0.18 mm at PL1 compared to larvae fed with diet D0 and D3 ( P > 0.05). However, in both experiments prawn larvae fed with the various diets showed significant differences in their length-at-stage ( P > 0.05), and the lowest growth and survival were obtained from diets comprising 3% and 5% bacterial biomass, which also gave the highest levels of ammonia-nitrogen in the culture water. Addition of more than 2% R. sulfidophilum biomass apparently deteriorates the quality of the culture water.     

几种养成虾的病毒携带状况检测

用组织病理学方法检测了深圳市及其临近地区养殖对虾携带病毒的情况。检查１１８尾外观正常的养殖对虾，包括斑节对虾，长毛对虾，近缘新对虾和刀额新对虾。检出带有斑节对虾杆状病毒包涵体的个体２０尾，约占１７％；带有对虾肝胰腺类细小病毒包涵体的个体４尾，约占３．３％。     

生物絮团在斑节对虾养殖系统中的形成条件及作用效果

为确定斑节对虾（Penaeus monodon）养殖系统中生物絮团形成所需的最适碳源及添加量,试验选取葡萄糖、蔗糖、糖蜜3种碳源按碳（C）与氮（N）比（C/ N）为20：1添加,再按日投饲量的0％、25％、50％、75％和100％添加蔗糖,以期获得其最适添加量。结果显示,添加3种不同糖作为碳源均可以显著提高斑节对虾的特定生长率（P ﹤0.05）,而添加蔗糖可以显著提高斑节对虾的成活率（P ﹤0.05）;与其他组相比,添加75％的蔗糖组可以显著提高斑节对虾的特定生长率和成活率（P ﹤0.05）,促进异养细菌和浮游植物的繁殖,水体氨氮（NH4-N）、亚硝酸氮（NO2-N）等有害物质的浓度较低,过多的添加蔗糖却会起到反作用。结果表明添加75％的蔗糖最适合生物絮团在斑节对虾养殖系统中形成,并促进斑节对虾的生长。     

斑节对虾体长与中肠腺细胞数相关性研究

通过现场测定养殖日龄为１６～１８ｄ的斑节对虾的体长、体重和全长,并应用石蜡切片技术对被测虾的中肠腺中部进行细胞计数,研究了斑节对虾体长与中肠腺细胞数的相关性。结果表明,对虾体长与中肠腺中部每个高倍视野的平均细胞数呈密切负相关（ｒ＝－０．９４７）,公式为：Ｃ＝３３０．３６２３２３ｅ－０．０８７３８４Ｌ。对虾体长与体重密切相关（ｒ＝０．９４８）,可用对数回归方程描述：ｌｇＷ＝－１．９２３５７８＋３．０８２８３２ｌｇＬ。对虾体长与全长密切相关（ｒ＝０．９９５）,其线性回归方程为：Ｙ＝－０．１８７１３７＋１．３１６７０５Ｘ。通过这些公式,中肠腺中部每个高倍视野的平均细胞数可由体长直接推算出或由体重和全长间接算出,使斑节对虾杆状病毒感染度的计算更加简单和准确。     

Cu2+和Cd2+对斑节对虾幼虾的毒性作用

研究了Cu^2 和Cd^2 对斑节对虾幼虾 的急性和亚急性毒性作用，获得了Cu^2 和Cd^2 的半致死浓度、安全浓度和累积量。     

Influence of ammonia on growth of Penaeus monodon Fabricius post-larvae

Abstract. Tiger shrimp, Penaeus monodon Fabricius, post-larvae (32·0±3mg, 1·43±0·03 cm) were exposed to control, 0·12, 0·60, 1·20 and 2·40mg/l ammonia-N (un-ionized plus ionized ammonia as nitrogen) which is equivalent to control, 6,32,63 and 126μg/1 NH₃-N (un-ionized ammonia as nitrogen) for 8 weeks in 25 ppt, as pH of 7·85–8·18 and 26–28°C by static renewal method. Growth in weight and length of the shrimps exposed to 1·20 and 2·40 mg/l ammonia-N were significantly lower (P < 0·05) than those exposed to control. The EC₅₀ (concentration that reduced growth by 50% of that of the controls) was 1·33 mg/l ammonia-N, 70 μg/l NH₃-N for weight gain, and 2·35 mg/1 ammonia-N, 123μg/l NH₃-N for length gain of P. monodon post-larvae. The'maximum acceptable toxicant concentration'(MATC) of ammonia-N and NH₃-N for P. monodon post-larvae was 0·60mg/l and 32μg/l, respectively after 6 weeks of exposure.