BAC end sequencing of Pacific white shrimp <Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>: a glimpse into the genome of Penaeid shrimp

Little is known about the genome of Pacific white shrimp (Litopenaeus vannamei). To address this, we conducted BAC (bacterial artificial chromosome) end sequencing of L. vannamei. We selected and sequenced 7 812 BAC clones from the BAC library LvHE from the two ends of the inserts by Sanger sequencing. After trimming and quality filtering, 11 279 BAC end sequences (BESs) including 4 609 paired- ends BESs were obtained. The total length of the BESs was 4 340 753 bp, representing 0.18% of the L. vannamei haploid genome. The lengths of the BESs ranged from 100 bp to 660 bp with an average length of 385 bp. Analysis of the BESs indicated that the L. vannamei genome is AT-rich and that the primary repeats patterns were simple sequence repeats (SSRs) and low complexity sequences. Dinucleotide and hexanucleotide repeats were the most common SSR types in the BESs. The most abundant transposable element was gypsy, which may contribute to the generation of the large genome size of L. vannamei. We successfully annotated 4 519 BESs by BLAST searching, including genes involved in immunity and sex determination. Our results provide an important resource for functional gene studies, map construction and integration, and complete genome assembly for this species.     

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。     

Effect of copper on the growth of shrimps Litopenaeus vannamei: water parameters and copper budget in a recirculating system

Shrimps( Litopenaeus vannamei) were intensively cultured in a recirculating aquaculture system for 98 days to investigate effects of 0.3 mg/L Cu on its performance, Cu budget, and Cu distribution. Shrimps in Cu-treated systems had greater mean final weight(11.10 vs 10.50 g), body length(107.23 vs 106.42 mm), survival rate(67.80% vs 66.40%), and yield(6.42 vs 5.99 kg/m 3), and lower feed conversion ratio(1.20 vs 1.29) than those in control systems but the differences were not significant. Vibrio numbers remained stable(10 4 –10 6 colony forming units/mL) in the rearing tanks of both control and treated systems. Total ammonium-N, nitrite-N, nitrate-N, pH, chemical oxygen demand, 5-day carbonaceous biochemical oxygen demand, and total suspended solids were similar in controls and treatments. Dissolved Cu concentration in the treated systems decreased from 0.284 to 0.089 mg/L while in the control systems it increased from 0.006 2 to 0.018 mg/L. The main sources of Cu in the treated systems were the artificially added component(75.7% of total input), shrimp feed(21.0%), water(2.06%), and shrimp biomass(1.22%). The major outputs of Cu occurred via the mechanical filter(41.7%), water renewal(15.6%), and draining of the sediment trap(15.1%). The foam fractionator removed only 0.69% of total Cu input. Harvested shrimp biomass accounted for 11.68% of Cu input. The Cu concentration of shrimps in the Cu-treated systems(30.70 mg/kg wet weight) was significantly higher than that in control systems(22.02 mg/kg). Both were below the maximum permissible concentration(50 mg/kg) for Cu in seafood for human consumption in China. Therefore, recirculating systems can be used for commercial on-growing of Litopenaeus vannamei without loss of shrimp quality, even in water polluted by 0.30 mg/L Cu. The mechanical filter is the main route for Cu removal.     

Effects of Hypoxia on Dopamine Concentration and the Immune Response of White Shrimp （Litopenaeus vannamei）

Effects of hypoxia on the dopamine concentration and the immune response of White Shrimp Litopenaeus vannamei were studied. The results showed that hypoxia had significant effects on the concentration of dopamine (DA) in the haemolymph, haemocyte count, phenoloxidase activity, phagocytic activity of haemocytes and bacteriolytic and antibacterial activity in the haemolvmph (P<0.05). The concentration of the dopamine in haemolymph reached its maximum in the 3.0 and 1.5mgL-1 DO groups at 12h and 6h, and then returned to normal after 24h and 12h, respectively. All immune parameters decreased with the reduction of dissolved oxygen. Total haemocyte count (THC), the hyaline cells and semi-granular cells in the 3.0mgL-1 DO group became stable after 12h,while granular cells did so after 24h, The THC and different haemocyte count (DHC) in the 1.5mgL-1 DO group became stable alter 24h. Phenoloxidase activity and bacteriolytic activity in the 3.0 and 1.5mgL-1 DO groups reached their stable levels after 24 h and 12 h respectively, while phagocytic activity and antibacterial activity became stable after 24 and 12, and 36 and 24 It, respec-tively. It was also indicated that the changes of dopamine concentrations in haemolymph, haemocyte count and phenoloxidase activ-ity were obviously related to the exposure time under hypoxic conditions.     

Growth and feed efficiency of juvenile shrimp Litopenaeus vannamei fed formulated diets containing different levels of poultry by-product meal

This feeding trial was conducted to evaluate the potential of poultry by-product meal (PBM) as a protein source in the culture ofLitopenaeus vannamei. Seven isonitrogenous and isoenergetic diets were formulated to near to commercial diet with about 40% protein and 7.5% lipid. Fish meal was replaced by 0, 30%, 40%, 50%, 60%, 70% and 100% of PBM (diets 1-7). The diet with 100% fish meal was used as a control (diet 1). Post-larvae were reared in an indoor semi-closed re-circulating system. Each dietary treatment was tested in 4 replicate tanks (260 L) of 40 shrimp, arranged in a completely randomized design. The shrimps were hand-fed for three times a day to near-satiation (0700, 1200 and 1800) for 60d. Percentage weight gain, survival, feed conversion ratio (FCR), protein efficiency ratio (PER) and body composition of shrimps were measured. There were no significant differences (P>0.05) in growth performance among shrimps fed diets 1-5 (0-60% fish meal replacement). However, shrimps fed diet 7 (100% fish meal replacement) had significantly lower (P<0.05) growth than those fed diets 1-5 (0-60% fish meal replacement). Shrimp fed diets 2-4 (30%-50% fish meal replacement) showed significantly higher growth than those fed diets 6 and 7 (70% and 100% fish meal replacement, respectively). Survival ranged from 94.7% to 100.0% and did not differ significantly (P>0.05) among different experimental diets. No differences in body composition were found among shrimps fed different diets. These results showed that up to 70% of fish meal protein can be replaced by PBM without adversely affecting the growth, survival, FCR, PER and body composition of Litopenaeus vannamei.     

L-精氨酸对凡纳滨对虾体液免疫因子的影响

用添加不同剂量L-精氨酸(L-arg)的饲料投喂凡纳滨对虾(Litopenaeus vannanei),测定其血淋巴液中一氧化氮(NO)含量以及一氧化氮合成酶(NOS)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)和超氧化物歧化酶(SOD)活力,并检测其对病原体的抗病力.结果显示:L-arg显著提高血淋巴液中NO含量,且变化趋势与NOS、AKP活性升高趋势相一致;中等剂量(质量分数为1%)L-arg显著提高血淋巴液中SOD和ACP活性,但高剂量(质量分数为1.5%)、低剂量(质量分数为0.5%)L-arg对血淋巴液中SOD(高剂量除外)和ACP活性影响不显著.副溶血弧菌人工感染后,中剂量L-arg组(1%)对虾死亡率最低,高剂量(1.5%)、低剂量组(0.5%)次之,对照组死亡率最高,各剂量L-arg组死亡率与对照组相比差异显著.表明饲料中添加适量L-精氨酸可使对虾体液免疫因子总体活力达到较高水平,L-精氨酸含量过高对对虾非特异免疫力的作用效果反而不佳.     

蛋白质营养对高密度养殖凡纳滨对虾生长与免疫力的影响

在高密度养殖条件下,进行单因素随机设计动物试验.用5种饲料(蛋白质水平31%、35%、39%、43%、47%,以A～E组表示)分别投喂平均体质量6.2 g+0.2 g、平均养殖密度3.1 kg/m3的凡纳滨对虾(Litopenaeus vannamei),探寻蛋白质营养对虾生长、免疫、水质、抗胁迫的影响特征.结果表明:(1)中高蛋白质水平具有显著促进对虾生长的效果,随着蛋白水平的提高,特定生长率先增加后降低,饲料系数正好相反,D组两指标最佳,分另q为110.98%和2.54;C、D、E组差异不显著.(2)中高蛋白质水平有利于提高对虾多数免疫指标的活力,血淋巴中血细胞浓度、T-AOC活力、POD活力、总蛋白含量、白蛋白、血蓝蛋白含量,随着蛋白质水平提高先增加后降低,前5指标含量均以D组最高,比A组显著提高16.8%～33.9%;而血蓝蛋白含量C组最高,比A组提高15.0%.(3)高蛋白质水平有利于提高对虾SOD活力,也显著增强抗低盐胁迫的能力,但同时极显著加大了水环境中氨氮和亚硝氮的污染.(4)在我国北方集约化高密度养殖条件下,凡纳滨对虾中后期生长阶段适宜的饲料蛋白质营养水平为39%～43%.     

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。     

凡纳滨对虾性别分化的初步研究

通过组织学连续切片与扫描电镜观察,研究凡纳滨对虾的性别分化。结果显示:凡纳滨对虾外部形态的性别分化最早发生在仔虾后第37天,此时在扫描电镜下可观察到雄性附肢的芽基,仔虾后第45天,光镜下可观察到交接器;仔虾后第24天出现原始生殖细胞(PGCS),位于背大动脉的两侧及围心腔膜的腹面,细胞个体及细胞核明显比周围的细胞大,具有强嗜碱性,细胞核圆形或椭圆形,核内染色质清晰可见,随着生殖腺的发育,在第43天出现生殖管,第55天生殖腺可区分为精巢和卵巢。     

纳米生态基在凡纳滨对虾养殖中的应用研究

研究了凡纳滨对虾集约化养殖池塘中挂设不同密度的纳米生态基对养殖水质及对虾生长的影响。试验共使用12口养殖池,生态基的挂设密度分别为0.029,0.043和0.058 m2/m3水体,以不挂设纳米生态基的养殖池为对照。结果表明,挂设纳米生态基能明显降低养殖水体的氨氮、亚硝氮和COD含量,挂设生态基处理组池塘水体中氨氮与对照组之间差异显著(P0.05),亚硝氮在试验第7 d后差异显著(P0.05);纳米生态基能较好的稳定池塘水体的DO水平,增氧时DO增加速度与挂设生态基的密度成正比,增氧关闭时DO减小速度与挂设生态基的密度成反比;生态基能提高凡纳滨对虾产量并有效降低其饵料系数,对虾产量分别比对照组高出0.8%,15.3%,3.2%,饵料系数分别降低17%,40%,26%。当对虾放苗密度为150尾/m2时,在本研究设计的梯度范围内,生态基的最佳挂设密度为0.043 m2/m3水体。