影响南美白对虾苗种质量的关键因素研究

南美白对虾是我国虾类养殖的主导品种,由于对虾养殖是一项风险高、利润大的系统工程,在众多风险因子中苗种质量又是养殖南美白对虾成功与否的首要限制因素,因此有效选择优质对虾苗种对我国南美白对虾的持续健康养殖有着重要意义。从育苗和放苗环节综述了影响南美白对虾苗种质量的关键因素,以期为对虾养殖生产中进行优质苗种选择和放养提供必要的参考依据。     

Effects of water exchange protocols on water quality,sedimentation rate and production performance of Penaeus monodon in earthen ponds

This study was carried out in farmers' fields to quantify the total water and consumptive water use in grow‐out culture of Penaeus monodon under recommended package of practice with two different water management protocols: T₁, with no water exchange and T₂, with regulated water exchange. Treatment‐wise estimated total water use, was 2.09 and 2.43 ha‐m 122 day^?1, while the computed consumptive water use index (m³ kg^?1 biomass) was 5.35 and 6.02 in T₁ and T₂ respectively. Lower rates of water exchange (T₂) showed significantly improved (P < 0.05) crop performance in terms of performance index (19.75 ± 0.75), production‐size index (74.1 ± 3.4), survival rate (80.13 ± 1.7%) and productivity (2.44 ± 0.08 t) over the zero water exchange. The shrimp pond water quality suitability index (WQSI) infers that regulated water exchange (T₂) improved the overall suitability of water quality for shrimp culture. WQSI up to 90 days of culture ranged between 7.5–9.0 in T₂, needs little management while in the last month of rearing, it was good with moderate management requirements. Treatment‐wise sediment load ranged between 50.4–56.3 m³ t^?1 shrimp biomass. High intensity of water exchange and low apparent feed conversion ratio influenced in lowering the sedimentation rate. Regulated water exchange protocol (T₂) performed well (higher net total water productivity and net consumptive water productivity) against no water exchange (T₁). A higher OV:CC ratio (ratio of the output value to the cost of cultivation) indicated that T₂ had a distinct edge over the T₁ protocol.     

斑节对虾细胞周期蛋白Y基因克隆与原核表达

为了研究细胞周期蛋白Y(cyclin Y)在斑节对虾(Penaeus monodon)卵巢发育中的作用,从斑节对虾转录组数据中筛选获得cyclin Y基因部分序列,采用SMART-RACE方法克隆得到斑节对虾细胞周期蛋白Y(Pm-cyclin Y)基因c DNA全序列。Pm-cyclin Y基因c DNA全长1576 bp,其中包含108 bp的5′非编码区(5′UTR)和439 bp的3′非编码区(3′UTR)以及1029 bp的开放阅读框(ORF),可编码342个氨基酸。生物信息学分析显示,其编码的氨基酸序列有1个保守的周期蛋白框(cyclin box)同源结构域(172~257 aa),预测的分子量约为37.6 k D,理论等电点6.64。实时定量PCR显示其m RNA在卵巢的表达量显著高于其他组织(P0.05);并且在卵巢5个不同发育期都有表达,在III期卵巢中的表达量最高。本研究通过原核表达方法对Pm-cyclin Y进行重组表达,为其蛋白质功能方面的研究奠定了基础。     

Enhancement of innate immune system,survival and yield in Penaeus monodon reared in ponds using Streptococcus phocae PI80

The objective of this study was to evaluate the effect of dietary and water supplementation of probiotic Streptococcus phocae PI80 on growth, immune response and feed utilization of tiger shrimp Penaeus monodon in earthen ponds. The probiotic bacterium S. phocae PI80 was cultured in large fermenter (50 L) by adding additional carbon source in the form of molasses and glucose along with yeast extract as nitrogen source to enrich S. phocae PI80 biomass. This enriched S. phocae PI80 was administered to shrimp in feed (6.5 × 10¹³ bacterial cells mL^?1) as well as in pond water (5 L/pond). Shrimp growth performance was significantly improved (P < 0.05) in 120 days culture when the average body weight of treated molasses + yeast extract (MY) (28.41 ± 0.874 g), glucose + yeast extract (GY) (27.013 ± 0.698 g) was significantly higher than control (23.63 ± 0.684 g). Food conversion ratio FCR was also found to be reduced significantly in ponds treated with probiotics when compared with control pond (1.89 ± 0.09). Vibrio and luminescent bacteria were found to be lower in the treatment receiving MY group, and we hypothesize that this may lead to greater shrimp survival. Furthermore, fermentation product of S. phocae PI80 added to pond water and feed additives enhanced the shrimp immune system. The results indicated that total haemocytes count (THC), phenoloxidase (PO) activity, NBT reductase assay and phagocytic activity significantly increased in shrimps treated with S. phocae PI80. Our study demonstrates that administration of S. phocae PI80 in the water and feed at 6.5 × 10¹³ colony‐forming units (CFU) mL^?1 bacterial cells induce immune modulation and enhances the immune ability of P. monodon in pond‐reared shrimp and increased the shrimp production.     

斑节对虾细胞周期蛋白B慢病毒载体的构建及对Hela细胞增殖影响

细胞周期蛋白B（Cyclin B）是影响卵母细胞发育成熟的重要基因。文章构建了PmCyB慢病毒表达载体,并和慢病毒包装复合物在293T细胞中包装成慢病毒颗粒。利用包装好的慢病毒颗粒,感染干扰了Cyclin B1基因的Hela细胞,研究了PmCyB对Hela细胞增殖的影响。结果表明,通过siRNA干扰细胞自身的Cyclin B1后,转染对照组病毒液的Hela细胞增殖减缓;同时,干扰后转入含有PmCyB重组慢病毒表达载体,Hela细胞增殖速度明显要高于转染对照组病毒液的Hela细胞的增殖速度。这一结果表明,PmCyB基因的过表达能补偿Cyclin B1缺失导致的细胞增殖减缓的现象,说明PmCyB基因具有细胞周期调控功能,是细胞增殖与分化的重要调控基因,且功能可能与人类的Cyclin B1相似。人类的Cyclin B1是卵母细胞发育成熟的重要基因。由此可以推断,PmCyB基因可能对斑节对虾（Penaeus monodon）卵巢发育成熟起着重要作用。     

Immunostimulatory effect of mangrove‐derived marine yeasts in Penaeus monodon

This study attempted to assess the efficacy of mangrove‐derived marine yeasts as a source of immuno‐stimulant in the tiger shrimp, Penaeus monodon. The shrimp was fed on the diets in supplementation with yeast biomass at 10% for a period of 15 days and then challenged with the pathogenic Vibrio cholerae. The animals were assessed for immune responses in terms of total haemocyte counts, phenol oxidase and endobiotics. Among the yeasts tested, Rhodotorula minuta was found to have high immunostimulatory effect in the shrimps especially after challenge with the pathogenic vibrio.     

中国对虾病原菌(鳗弧菌)的研究

1987年7～10月福建省平谭县对虾养殖场发生了一起严重的流行病。病虾活动力减弱,食欲下降,个体消瘦,头胸甲心区附近呈白色或浅桔红色,血淋巴液稀薄、混浊、不能凝固。因病虾步足、游泳肢及尾扇呈鲜红色,被称为“红腿病”。死亡率可高达90％以上。从五只垂死病虾的心脏血淋巴液中分离到细菌作纯焙养,经肌肉注射和浸泡的人工感染方法,得到了与虾池中患病虾相同的症状。菌原菌是革兰氏阴性短杆鼠。以极生单鞭毛运动。发酵葡萄糖,产酸不产气。氧化酶、过氧化氢酶阳性。能还原硝酸盐成亚硝酸盐。精氨酸脱羧;鸟氨酸、赖氨酸不脱羧。V.P.阳性.在42℃中不能生长。不能在无盐和含有8％NaCl 的胨水中生长,但在含有3％和6％NaCl的胨水中生长良好。对弧菌抑制剂0/129敏感。经鉴定为鳗弧菌(Vibrio anguillarum Bergman)。     

Pre-exposure to infectious hypodermal and haematopoietic necrosis virus or to inactivated white spot syndrome virus (WSSV) confers protection against WSSV in Penaeus vannamei (Boone) post-larvae

Larvae and post-larvae of Penaeus vannamei (Boone) were submitted to primary challenge with infectious hypodermal and haematopoietic necrosis virus (IHHNV) or formalin-inactivated white spot syndrome virus (WSSV). Survival rate and viral load were evaluated after secondary per os challenge with WSSV at post-larval stage 45 (PL45). Only shrimp treated with inactivated WSSV at PL35 or with IHHNV infection at nauplius 5, zoea 1 and PL22 were alive (4.7% and 4%, respectively) at 10 days post-infection (p.i.). Moreover, at 9 days p.i. there was 100% mortality in all remaining treatments, while there was 94% mortality in shrimp treated with inactivated WSSV at PL35 and 95% mortality in shrimp previously treated with IHHNV at N5, Z1 and PL22. Based on viral genome copy quantification by real-time PCR, surviving shrimp previously challenged with IHHNV at PL22 contained the lowest load of WSSV (0-1x10(3) copies microg-1 of DNA). In addition, surviving shrimp previously exposed to inactivated WSSV at PL35 also contained few WSSV (0-2x10(3) copies microg-1 of DNA). Consequently, pre-exposure to either IHHNV or inactivated WSSV resulted in slower WSSV replication and delayed mortality. This evidence suggests a protective role of IHHNV as an interfering virus, while protection obtained by inactivated WSSV might result from non-specific antiviral immune response.     

Development of a monoclonal antibody‐based flow‐through immunoassay (FTA) for detection of white spot syndrome virus (WSSV) in black tiger shrimp Penaeus monodon

A flow‐through immunoassay (FTA), an improved version of immunodot, was developed using a nitrocellulose membrane baked onto adsorbent pads enclosed in a plastic cassette to detect white spot syndrome virus (WSSV) in shrimp. Sharp purple dots developed with WSSV against the white background of the nitrocellulose membrane. The detection limits of WSSV by the FTA and immunodot were 0.312 and 1.2 μg mL^?1 crude WSSV protein, respectively. The FTA could be completed in 8–10 min compared with 90 min for immunodot. The FTA was 100 times more sensitive than 1‐step polymerase chain reaction (PCR) and in between that of the 1‐ and 2‐step PCR protocol recommended by the Office of International Epizootics (OIE). In experimental, orally infected shrimp post‐larvae, WSSV was first detected 14, 16 and 18 h post‐infection (hpi) by FTA, immunodot and one‐step PCR, respectively. The FTA detected WSSV 2 and 4 h earlier than immunodot and one‐step PCR, respectively. The FTA was more sensitive (25/27) than one‐step PCR (23/27) and immunodot (23/27) for the detection of WSSV from white spot disease outbreak ponds. The reagent components of the FTA were stable giving expected results for 6 m at 4–8 °C. The FTA is available as a rapid test kit called ‘RapiDot’ for the early detection of WSSV under field conditions.     

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.