1993～1994年山东省中国对虾育苗期病毒感染的流行病学调查

１９９３年和１９９４年育苗期，在山东省十余家育苗场采集了亲虾、受精卵、无节幼体、蚤状幼体、糠虾幼体和仔虾等样品共５６份。组织病理学切片观察说明，所有样品均未检查出对虾暴发性流行病病原ＨＨＮＢＶ的病理变化；ＨＰＶ的病理变化主要在亲虾、糠虾幼体和仔虾中检出，蚤状幼体有ＨＰＶ感染的可疑情况。１９９４年，５．５％的糠虾幼体和５．１％的仔虾检出有ＨＰＶ的感染。在糠虾和仔虾阶段，海捕亲虾苗种的ＨＰＶ带毒率为４．０％，越冬亲虾苗种的带毒率为６．２％。ＨＰＶ和ＨＨＮＢＶ的单克隆抗体ＥＬＩＳＡ检测方法表明有３个样品为可疑的ＨＨＮＢＶ阳性；有１２．５％～２５％的糠虾期育苗池和７０％的仔虾期育苗池存在有ＨＰＶ的感染。     

首例斑节对虾越冬亲虾发生杆状病毒的皮下及造血组织坏死症研究

湛江长洪对虾苗种实验场的斑节对虾转入越冬池后３～１０ｄ内发生暴发性死亡，症状与北方地区杆状病毒的皮下及造血组织坏死症一致。病理学研究表明，所有发病虾样的皮下组织、造血组织、结缔组织、肝胰腺血窦、淋巴器官等细胞核均存在严重的ＨＨＮＢＶ病灶，只有２０％的发病对虾肝胰腺中观察到轻微感染的ＭＢＶ包涵体。ＨＨＮＢＶ单抗的酶联免疫染色，可对ＨＨＮＢＶ的病灶产生特异性着色，对虾组织结构和ＭＢＶ包涵体均不着色。在山东培育三代并养殖１６个月的斑节对虾中也观察到与湛江发病的越冬亲虾相同的情况。以上说明，近年来在广东一带ＨＨＮＢＶ是值得重视的病原，其危害可能重于ＭＢＶ。对于发生过对虾暴发性流行病的群体来说，不宜用来作为亲虾。     

放流增殖对虾病毒检测报告

为了解黄海北部中国对虾放流增殖回捕率下降原因 ,1 995～ 1 998年我们对在黄海北部中国对虾 (Ｐｅ ｎａｅｕｓｃｈｉｎｅｓｅ)放流的仔、幼虾进行了病毒检测。现将检测结果报告如下。1　检测方法样品分别取自育苗室放流用的 1ｃｍ仔虾、暂养期内及放流入海时的 2～ 3ｃｍ仔、幼虾和第一、第二航次调查的幼虾 ,体长 5～ 7ｃｍ。用两种方法测定病毒感染。用切片镜检方法检出的是发病个体。用核酸探针方法检出的称皮下及造血组织坏死杆状病毒 (ＨＨＮＢＶ)分四级 ,“十”指ＨＨＮ ＢＶ阳性 ,阳性弱 ,达到检测灵敏度 ;“ ”指阳性 ,较弱 ,易于…     

中国对虾杆状病毒垂直传播途径的初步探讨

中国对虾杆状病毒垂直传播途径的研究对切断病毒的传播途径，培育健康的对虾种质资源，具有重要的理论和实践意义。本文利用电镜技术，对中国对虾亲虾的卵巢，卵细胞和无节幼体，溲状幼体，糠虾，仔虾，幼成虾进行病毒检测，结果发现卵巢和卵细胞中有似病毒粒子，无节幼体，糠虾，仔虾，幼成虾有杆状病毒感染，并初步探讨了中国对虾杆状病毒垂直传播的可能性。     

中国对虾聚缩虫病的防治

中国对虾幼体的聚缩虫病是对虾育苗中常见的、危害较大的虾病，它感染速度快，几天之内便可使培育池中的对虾幼体８０％以上感染，如不及时防治，可导致育苗失败。 一、病原体 对虾幼体聚缩虫病的病原体常见种是树状聚缩虫，属原生动物门、纤毛虫纲、缘毛目、钟虫科。聚缩虫寄附于中国对虾蚤状幼体、糠虾幼体、仔虾及成虾的体表和鳃。 二、发病过程及症状 对虾蚤状幼体、糠虾幼体及仔虾都容易感染聚缩虫，尤以蚤状幼体、糠虾幼体的聚缩虫病危害最大。１９９９年，大连市金州区北海村育苗室对虾蚤状幼体第Ⅲ期时，５月１４日发现第２０号培育…     

中国对虾皮下及造血组织坏死杆状病毒（HHNBV）DNA探针的制备及应用

ＨＨＮＢＶ是１９９３年中国对虾暴发性流行病的主要病原之一。从纯化的病毒中提取ＤＮＡ，用ＥｃｏＲｌ限制性内切酶酶切成ＤＮＡ片段，与ＰＵＣ１８体外重组，建立ＨＨＮＢＶＤＮＡ文库。从中筛选出三个重组质粒（５＃、８＃、９＃），它们所插入的片段大小分别为：２．８Ｋｂ、０．８Ｋｂ、１．６Ｋｂ，它们分别用光敏生物素标记成探针，与感染ＨＨＮＢＶ的病虾ＤＮＡ呈阳性反应，以此利用制备的ＨＨＮＢＶＤＮＡ探针的高敏感性和特异性来检测虾病。     

用于PCR检测对虾皮下及造血组织坏死杆状病毒(HHNBV)的一种快速提取DNA方法

使用采样液ＳＥＭＰ－Ｔｒｉｓ保存人工感染ＨＨＮＢＶ（ＷＳＳＶ的一个分离株）的中国对虾组织，然后分别用酚抽提法、玻璃乳（Ｇｌａｓｓ Ｍｉｌｋ）回收法、硝酸纤维素膜结合法和煮沸－乙醇沉淀法提取ＤＮＡ。应用ＨＨＮＢＶ引物对提取的ＤＮＡ进行ＰＣＲ扩增，使用琼脂糖凝胶电泳对扩增产物进行鉴定和检测。通过比较表明，煮沸－乙醇沉淀法是一种最快速、简便的从采样液ＳＥＭＰ－Ｔｒｉｓ保存的病虾组织中制备ＰＣＲ模板的方法     

对虾中子辐照刺激增产技术研究

采用１４ＭＥＶ快中子对中国对虾、长毛对虾、墨吉对虾的亲虾、受精卵、无节幼体和仔虾进行一定剂量的辐照，可以提高产卵量、好卵率、孵化率、成活率，促进对虾的生长发育，增强其抗病能力，提高单位面积的成虾产量，改善商品虾的品质。     

白斑综合症病毒（WSSV）对中国对虾卵及各期幼体人工感染的试验研究

用感染白斑综合症毒病的中国对虾头胸甲，对健康的仔虾进行了人工投喂感染实验，同时从病虾中分离出病毒悬液作为毒种，无节幼体，蚤体 幼体和糠虾体进行不同温度条件下的人工浸浴实验，结果表明，卵和无节幼体，蚤状幼体，糠虾幼对病毒悬液不敏感,闰理切片观察未见病毒粒子，光镜病理切片可观空到鳃上皮，前肠上皮有包涵体样病变，同时在肝胰组织中发现了肝胰腺细小病毒（HPV）包涵体。     

我国对虾健康养殖技术

一、对虾健康养殖管理的要求我国目前在对虾病毒性暴发流行病防治研究方面已取得明显进展 ,虾病防治的实践 ,也使人们认识到健康养殖管理 ,不但可以避免暴发性流行病发生 ,而且它也是对虾养殖可持续发展的重要环节。通常我们认为健康养殖管理 ,可概括为如下主要内容。１．培育和使用健康虾苗 :其主要技术关键是在苗种培育中 ,避免使用携带可引起暴发性流行病病原的亲体。当前主要是检疫ＨＨＮＢＶ（亦称白斑综合症病毒）及ＭＢＶ（斑节对虾杆状病毒）等病毒。对亲体的遗传特性应作严格选择。例如育苗单位使用人工培育亲体时 ,应避免使用同一…     

单克隆抗体酶联免疫技术检测对虾皮下及造血组织坏死病的病原及其传播途径

用单克隆抗体ＥＬＩＳＡ技术对１９９４年５月～７月自山东和辽宁采集的虾池和海区材料，包括对虾、浮游生物、小型甲壳类生物、鱼类、底泥及饲料等共１９２个样品进行了对虾暴发性流行病病原ＨＨＮＢＶ的检测。结果表明，对虾中的ＨＨＮＢＶ阳性与虾池发病情况基本相符，用单抗ＥＬＩＳＡ方法可提前２０～４０ｄ对虾池的发病可能性作出预报；桡足类浮游生物的带毒率高于对虾，且其阳性的出现早于对虾，沿岸海区的桡足类浮游生物有较高的阳性率，它可能是对虾暴发性流行病的主要病原携带者；部分地区的卤虫也带有ＨＨＮＢＶ，同时还从糠虾等小型甲壳类生物中检出了病原。     

1994年浙江省对虾暴发性流行病病原及传播途径的初步调查

用Ｔ—Ｅ染色、单克隆抗体的ＥＬＩＳＡ现场诊断、组织切片和Ｈ—Ｅ染色和电镜观察等不同方法对１９９４年浙江省乐清和温岭两地区虾池及海区样品进行了现场和实验室检查。结果说明，１９９４年浙江省对虾暴发性流行病实质上是杆状病毒性的皮下及造血组织坏死，病原为ＨＨＮＢＶ。检查到的ＨＰＶ不是这场暴发病的病原。对非对虾类生物和对虾鲜活饲料的单抗ＥＬＩＳＡ检测表明，海区张网饲料可能是１９９４年浙江对虾暴发性流行病的主要传染源，病虾池中除对虾外，其它甲壳类生物在该病的传播上可能也起到一定的作用。这些生物除了可能传播ＨＨＮＢＶ外，还可能传播ＨＰＶ。     

对虾皮下及造血组织坏死杆状病毒单克隆抗体的研制

用经密度梯度离心纯化的对虾皮下及造血组织坏死杆状病毒（ＨＨＮＢＶ—９３７）注射Ｂａｌｂ／ｃ小鼠，取其免疫后的脾细胞与ｓｐ２／０骨髓瘤细胞进行融合。经筛选与克隆，获得７株抗ＨＨＮＢＶ的单克隆杂交瘤细胞株。其中Ｆ９株的染色体数为８７，ＥＬＩＳＡ测定培养上清中单抗滴度为１∶８，腹水为１∶１０００。患有杆状病毒的皮下及造血组织坏死症的对虾组织切片的单抗酶联免疫染色，表明Ｆ９株单抗只与ＨＨＮＢＶ病灶发生特异性结合，与对虾组织结构不发生特异性反应。将其用于现场样品的检测研究表明，该抗体适于用来进行对虾暴发性流行病的病原检测。     

ELECTRICITY AND PROPANE CONSUMPTION OF A SHRIMP HATCHERY IN THE STATE OF BAJA CALIFORNIA SUR,MEXICO

The consumption of electricity and propane for producing larvae and postlarvae of the whiteleg shrimp (Litopenaeus vannamei) was examined in a commercial shrimp hatchery on the Baja California Peninsula of Mexico. Between January and August 2005, 6 × 10⁶ postlarvae (average age PL16) were produced from 1.43 × 10⁹ nauplii. During that production period, the hatchery used 2.48 × 10⁹ kcal of fossil fuel energy (30% for electricity, 70% for liquid propane), which was equivalent to 16% of the operating costs. Electricity was used mainly for larval and postlarval rearing (36% and 10%) and microalgae culture (27%). During this production period, 1.47 × 10⁹ kcal of propane were consumed for heating daily more than 600 m³ seawater daily. Of that total, 33% was used for broodstock maintenance, 65% for larvae culturing, and 2% for postlarvae rearing. With increasing costs for shipping postlarvae to the mainland Mexican coastal areas (15% of operating costs) and the need to remain competitive with hatcheries in the coastal areas of Sonora and Sinaloa, alternative energy sources to reduce overall operational costs in hatcheries of Baja California Sur are discussed.     

大弹涂鱼仔鱼的摄食、生长和成活的研究

本文主要论述大弹涂鱼仔鱼阶段有关摄食和生长发育，以及不同饵料对仔鱼生长和成活的影响。结果表明，孵化后3—5日龄即仔鱼危险期，正处于内源性(包括混合性)营养转入外源性营养。颗粒有机碎屑是仔鱼的开口饵料，培育期间以底栖硅藻分解后形成的颗粒有机碎屑为饵料，其后期仔鱼成活率高达42．04％。6日龄起逐渐加投桡足类幼体和成体，其16日龄后期仔鱼平均全长可达6．15mm，平均体重1．24mg．     

Genetics of the resistance to hypoxia in postlarvae and juveniles of the Pacific white shrimp Penaeus (Litopenaeus) vannamei (Boone 1931)

The genetics of resistance to hypoxia was studied in shrimp using two approaches, heritability estimation and heterosis estimation. The heritability of resistance to hypoxia was evaluated in two populations of shrimp postlarvae families produced at different times, and heterosis was estimated through the reciprocal crosses of two genetically distinct shrimp populations at postlarvae and juvenile. Heritabilities of resistance to hypoxia in postlarvae were large and out of the range for the heritability (1.73±0.09, 1.35±0.14) regardless of population evaluated. When covariates (survival and mean total weight to postlarvae 15 days old) were introduced in the analyses to correct for effects during rearing of the larvae, the heritabilities did not change substantially for the first population, but were decreased for the second population depending on the covariate used for correction (1.15±0.10 and 1.08±0.11 for the covariates survival and total weight respectively). Those large heritabilities could be explained by the presence of confounded common environmental variance in the estimated genetic variance (maternal effects), or by dominance and epistatic effects being an important part of the estimated genetic variance. Heterosis analyses dissecting maternal effect indicated that both effects, maternal and dominance, are involved in the resistance to hypoxia at postlarvae, and that although maternal effects decreased at juvenile stage, the trait was still characterized by presenting a large heterosis value (62–65% for postlarvae and 36% for juveniles).     

A rapid non-enzymatic method of DNA extraction for PCR detection of white spot syndrome virus in shrimp

The present study describes a simple method of extraction of white spot syndrome viral DNA (WSSV) from infected shrimp for the polymerase chain reaction (PCR) detection of WSSV. The DNA preparation using this method was found to be free from the host DNA, RNA and protein, and is suitable for different PCR protocols such as single‐step PCR, nested PCR and single‐tube semi‐nested PCR. This method of extraction has worked successfully for extracting the WSSV‐DNA from different organs (haemolymph, eyestalk, carapace, head muscle, heart, gills, appendages, heptopancreas, stomach, intestine, abdominal muscle and tail muscle) of WSSV‐infected adult shrimp, and WSSV‐infected larvae and postlarvae.     

Characteristics and pathogenicity of a Vibrio campbellii-like bacterium affecting hatchery-reared Penaeus indicus (Milne Edwards, 1837) larvae

In the summer months, epizootics occurred regularly among larvae of Penaeus indicus (Milne Edwards) in a hatchery located at Narakkal near Cochin, India. The clinical signs of infected larvae were expansion of chromatophores. opaqueness of body with twisting and degeneration of appendages. Bacteria were isolated from the infected larvae. These bacteria had morphological, physiological and biochemical characteristics of the genus Vibrio, but these properties did not correspond with those of known Vibrio species. The Vibrio was most related to Vibrio campbellii according to DNA-hybridization experiments. By pathogenicity experiments, the present Vibrio isolate was proved to be pathogenic to larvae, postlarvae and adult shrimp. Shrimp larvae challenged by immersion showed significant mortalities. The bacterial isolate failed to produce the infection in adult shrimp after oral inoculation but it caused significant mortality in adults after intramuscular injection. LC⁵⁰ and LD⁵⁰ values of the Vibrio isolate were determined for larvae and adult shrimps, respectively, at different time intervals.