Valine requirement of postlarval tiger shrimp, Penaeus monodon Fabricius

The valine requirement of juvenile tiger shrimp, Penaeus monodon Fabricius, was determined. Shrimp postlarvae, PL20, with a mean weight of 14 mg, were randomly distributed in 36 oval 40-L capacity fibreglass tanks at 10 shrimp per tank in a flow-through seawater system and reared for 8 weeks. Postlarvae were fed amino acid test diets containing 400 g kg^?1 protein with casein and gelatine as intact sources of protein. Crystalline L-amino acids were supplemented to simulate the amino acid profile of the shrimp muscle except valine. Valine was added in graded levels to obtain 7, 10, 13, 16, 19 and 22 g kg^?1 of the diet or 18, 25, 33, 40, 48 and 55 g kg^?1 of dietary protein. At termination of the feeding experiment, growth and survival were determined and nutritional deficiency signs noted. The relationship between weight gain and dietary valine level was analysed by the broken-line regression method to derive the valine requirement. The dietary valine requirement of Penaeus monodon postlarvae was found to be 13.5 g kg^?1 of the diet or 34 g kg^?1 of dietary protein. This value was lower than the level found in the shrimp tissue.     

斑节对虾4个不同群体建立家系的生长及成活

以泰国（T）、非洲（A）和三亚（S）野生斑节对虾（Penaeusmonodon）群体,以及人工养殖斑节对虾群体（F）为父、母本建立家系。选择9个发育时间相近和较有代表性的家系,对其生长和饲料利用进行了为期30d的养殖试验。结果发现,大体上母本含三亚斑节对虾基因的家系成活率较高,其余较低,差异显著（P〈0．05）;母本含三亚斑节对虾基因的家系生长较差,其余较好（除T×T外）且差异极显著（P〈0．01）,A×A家系增重率是S×S的2倍;饵料系数没有显示明显的规律性。     

Probiotic bacteria effects on growth and bacterial composition of black tiger shrimp (Penaeus monodon)

The effects of Bacillus probiotic forms on growth performance and microbiota were examined in black tiger shrimp (Penaeus monodon). Shrimp were fed for 120 days with one of three diets; a control diet alone or supplemented with either probiotics consisting of live‐sprayed Bacillus (LS) or freeze‐dried Bacillus (FD). Culturable bacteria counts in the hepatopancreas, intestine and culture water of the three treatments were relatively constant during the experiment. Bacillus numbers in the hepatopancreas and intestines of probiotic‐treated groups increased significantly (P < 0.05) after 30 treatment days and remained relatively constant afterwards while no appreciable changes occurred in the control. Shrimp supplemented with LS and FD contained significantly lower concentrations of Vibrio (P < 0.05) in the hepatopancreas, intestine and culture water, compared to those in the control group. Shrimp treated with LS and FD probiotic bacteria showed a significant improvement (P < 0.05) in specific growth rate (SGR) and feed efficiency (FE), compared to those of control group. Probiotic bacteria have the potential to enhance shrimp culture by reducing Vibrio and improving growth performance.     

Fusarium incarnatum isolated from black tiger shrimp, Penaeus monodon Fabricius, with black gill disease cultured in Vietnam

Fusarium incarnatum was isolated from gill lesions of cultured black tiger shrimp, Penaeus monodon, in every crop during 2000-2002 in Nghe An province, Vietnam. Infected shrimps showed typical signs of black gill disease and mortalities about a month prior to harvest. Detailed morphological examinations, as well as molecular phylogenic analyses based on partial nucleotide sequences of ribosomal DNA, were made on the isolates. An artificial infection of kuruma prawn, Penaeus japonicus, using two selected isolates was also conducted and their pathogenicity determined.     

Spermatophore cryopreservation and artificial insemination of black tiger shrimp, Penaeus monodon (Fabricius)

To develop an appropriate cryopreservation protocol for spermatophores of black tiger shrimp, Penaeus monodon, three cryoprotectants (dimethyl sulphoxide (DMSO), methanol (MeOH) and ethylene glycol (EG)) at two concentrations (5% and 10%) were examined. Artificial implantation of spermatophores was also carried out to assess the fertilizing ability of fresh and post‐thaw spermatophores. Spermatophores were collected during consecutive regenerations (15‐day intervals) and assessed for qualitative and quantitative changes and also for fertilizing ability by implantation. The mean fertilization rate for artificial insemination using post‐thaw spermatophore was 79.9±3.7%, lower than the fertilization rates observed for artificial implantation using fresh spermatophore and natural mating. Mean hatch rates for fresh spermatophore, frozen‐thawed spermatophore and natural mating were 88.8±0.6%, 87.8±0.4% and 88.3±0.5%, respectively; and there was no difference among the three groups. The mean fertilization rate of spermatophores collected during the first stripping was higher (90.6±0.6) than during the second stripping (85.7±2.6), but the mean hatch rate was not different between the two strippings. The highest mean sperm viability (79.7±0.4%) was obtained from DMSO (5%), with no survival observed in the 10% MeOH treatment. Spermatophore weight, total sperm count and percentage of abnormal sperm were not different between spermatophores collected at the first and second stripping. This is the first study to report high fertilization and hatch rates from cryopreserved spermatophore using artificial implantation of spermatophore before spawning.     

The effectiveness of a commercial microbial product in poorly prepared tiger shrimp, Penaeus monodon (Fabricius), ponds

The efficacy of a commercial microbial product was tested in commercial tiger shrimp, Penaeus monodon (Fabricius), ponds for one culture period in Kuala Selangor, Malaysia. Four ponds with replicates for treatment and control were used. The pond bottom was dried but the organic sludge was not removed as normally practised in pond preparation. The ponds were stocked with 15 post‐larvae at the rate of 31.m^?2. Physical, chemical and biological parameters of the pond were analysed every 2 weeks during the culture period. Water quality parameters remained within the optimum range for shrimp culture except for ammonia‐nitrogen being significantly higher in control ponds and silica in treated ponds. Benthic organisms were not found in any of the ponds. The average counts of different bacteria were not significantly higher in treated ponds than control. Because of poor health, the shrimp were harvested earlier (72 days) than the usual 120 days. An average of 875.60 ± 67.00 kg shrimp ha^?1 was obtained in treated ponds with a feed conversion ratio (FCR) of 1.57 ± 0.10 and survival rate of 42.35 ± 5.37% compared with 719.50 ± 130.94 kg shrimp ha^?1, 2.99 ± 0.70 and 21.25 ± 3.26%, respectively, in control ponds. Neither the microbial product nor the frequent water exchange was effective in overcoming the problems caused by the poor pond bottom.     

斑节对虾基因组微卫星分离及其序列特征研究

采用(CA)_(12)(AG)_(12)及(TA)_(16)生物素标记探针及磁珠富集法构建了斑节对虾Penaeus monodon基因组微卫星富集文库。随机挑选254个克隆进行PCR筛选,得到51个候选克隆(20．1%)。其中,32个克隆来源于CA-文库,另19个克隆来源于AG-文库。测序发现48个克隆含有微卫星重复单元,通过序列比对,最终获得40个具有特异微卫星序列的阳性克隆。微卫星(GA/CT)_N及(CA/GT)_n 2碱基重复序列分别占所有分离的微卫星数目的20．7%及60．4%。此外,还检测到其它多种微卫星重复类型,如(AT)_n、(GC)_n、(TGG)_n、(AAG)_n、(AAT)_n、(GAA)_n、(GTGC)_n、(GCGT)_n、(GGTTA)_n、(GTGCGT)_n,占检测到的微卫星数目的18．9%。获得的微卫星序列中属于完全型序列的有76条(68．5%),不完全型序列的有22条(19．8%),另有13条属于复合型序列(11．7%)。微卫星(GT/CA)_n 2碱基重复次数(3～52次)要远大于(GA/CT)_n 2碱基次数(3～27次)。获得的微卫星序列长度大小范围为129～601 bp,平均为286 bp。研究为进一步开展斑节对虾分子育种及资源评价分析提供了基础资料。     

Mycoflora of the 'green water' culture system of tiger shrimp Penaeus monodon Fabricius

This study was conducted to quantify and characterize the mycoflora associated with the ‘green water’ culture system of Penaeus monodon. Samples of water, tilapia gut and mucus, and shrimp hepatopancreas from three shrimp farms were collected during 15, 30, 45 and 60 days of culture (DOC). Results showed that high fungal loads were observed in tilapia gut (total: 117–1352 colony forming unit (CFU) 5 cm hind gut^?1; yeasts: 0–136 CFU 5 cm hind gut^?1) and mucus (total: 12–311 CFU (5 cm²)^?1; yeasts: 0–88 CFU (5 cm²)^?1), while minimal fungal populations were observed in water samples (total: 0–110CFU mL^?1; yeasts: 0–5 CFU ml^?1). Shrimp hepatopancreas harboured a very low number of filamentous fungi (0–27 CFU 0.1 g^?1) and yeasts (0–7CFU 0.1 g^?1) especially at 60 DOC. The filamentous fungal isolates were dominated by Penicillium and Aspergillus species, while the yeast populations were dominated by Rhodotorula and Saccharomyces species. The dominance of these fungi on tilapia mucus and gut and their presence in the rearing water might play an important role in the overall mechanisms involved in the control of luminous Vibrio in the ‘green water’ grow‐out culture of P. monodon.     

Dietary biofloc supplementation in black tiger shrimp,Penaeus monodon: effects on immunity,antioxidant and metabolic enzyme activities

A 60‐day indoor experiment was conducted to study the effect of dietary supplementation of biofloc on metabolic enzyme activities and immune responses in Penaeus monodon juveniles. Biofloc developed in indoor fibreglass‐reinforced plastic (FRP) tanks (1000 L) was used as dietary supplement in P. monodon (2.90 ± 0.10 g) reared in 1000‐L FRP tanks. Graded level of dried biofloc was included in shrimp basal diets, 0% (control, B0), 4% (B4), 8% (B8) and 12% (B12). The level of metabolic enzymes like malate dehydrogenase (MDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) was not significantly different with control up to 8% dietary supplementation. A higher level of total haemocyte count (THC) was noticed in B8 (22.16 ± 2.17 × 10⁶ cells mL^?1) and B4 (21.11 ± 0.56 × 10⁶ cells mL^?1) compared with control, C (14.61 ± 2.74 × 10⁶ cells mL^?1). Biofloc‐supplemented groups recorded significantly higher (P < 0.05) serum SOD and catalase activity (P < 0.01) in comparison with control. The groups fed with 4% dietary biofloc supplement recorded highest relative percentage survival (RPS), 45% after challenge with Vibrio harveyi followed by 36% and 27% RPS in B8 and B12 groups. Based on these results, it can be concluded that supplementation of biofloc even at 4% level in the feed improves immune responses and metabolic activities in black tiger shrimp juveniles.     

Dietary potassium requirement of juvenile grass shrimp Penaeus monodon

ABSTRACT: A feeding trial was conducted to estimate the minimal dietary potassium (K) requirement for juvenile grass shrimp Penaeus monodon . Purified diets with seven levels (0, 0.3, 0.6, 0.9, 1.2, 1.5, 2.0 g/100 g) of supplemental potassium were fed to P. monodon (mean initial weight 0.75 ± 0.01 g) for 8 weeks. Each diet was fed to three replicate groups of shrimp. The rearing water contained 360 mg/L potassium. Weight gain and protein efficiency ratio (PER) of shrimp improved as dietary potassium supplementation level increased up to 1.5 g K/100 g diet and thereafter declined significantly ( P < 0.05). Feed efficiency (FE) of shrimp also showed a similar trend. However, whole body K concentration of the shrimp kept an increasing trend up to 2.0 g K/100 g diet. Analysis of the weight gain (percent) and PER of the shrimp by polynomial regression indicate that the minimal dietary potassium requirement in growing P. monodon is approximately 1.2 g/100 g.     

Effects of biofloc under different carbon sources and protein levels on water quality,growth performance and immune responses in black tiger shrimp Penaeus monodon (Fabricius, 1978)

A 75‐day 2 × 3 factorial experiment was conducted to evaluate the effect of two levels of dietary protein (32 and 40%) and two different carbon sources (rice flour–R and molasses‐M), and without carbohydrate (control–C) in black tiger shrimp Penaeus monodon juveniles (3.37 ± 0.04 g) at 100 nos. m^?3 in 100 L fibre reinforced plastic tanks. Biofloc volume and total suspended solid were higher in molasses added groups (32 + M and 40 + M) followed by rice flour (32 + R and 40 + R) and controls (32C and 40C). Molasses and rice flour addition significantly reduced (P < 0.01) the total ammonia–N compared to controls. The highest Vibrio, Bacillus and Lactobacillus counts were recorded in 40 + M, 32 + M and 32 + R respectively. Among the treatments, significantly higher (P < 0.01) final body weight was recorded in 40 + R (8.5 ± 0.3 g), 40 + M (7.8 ± 0.3 g) and 32 + R (7.5 ± 0.4 g) compared to control groups, 32C (6.1 ± 0.3 g), 40C (6.4 ± 0.3 g) and molasses added group, 32 + M (5.7 ± 0.4 g). Rice flour supplementation significantly increased (P < 0.01) the total haemocyte count (×10⁶ cells mL^?1) in 32 + R (45.7 ± 3.7) and 40 + R (44.3 ± 3.1) compared to controls, 32C (27.3 ± 3.4) and 40C (25.8 ± 0.9). Similarly, higher superoxide dismutase, catalase, serum protein and glucose were recorded in the rice flour added groups, 40 + R followed by 32 + R. Among the treatments, the highest level of prophenoloxidase (OD 490 nm, 0.3 ± 0.0) and survival after challenge with Vibrio harveyi (55.6%) was observed in 32 + R. The study elucidates that rice flour addition produces optimum level of biofloc with better growth and immune responses compared to molasses and control. Furthermore, rice flour addition at 32% protein level could replace 40% protein feed.     

斑节对虾血淋巴细胞对鳗弧菌的清除作用

通过检测注入斑节对虾(Penaeus monodon)血淋巴内的鳗弧菌(Vibrio anguillarumH)浓度、血淋巴细胞浓度和血淋巴细胞的组成变化,研究斑节对虾血淋巴细胞对进入体内细菌的清除作用。实验显示,斑节对虾能够迅速清除注入其体内的鳗弧菌。注射鳗弧菌悬液(107~108CFU/mL)5 min后,血淋巴中可检测到的鳗弧菌浓度是(1.4±0.6)×106CFU/mL,而2 h后所检测到的鳗弧菌的浓度仅相当于前者的3.1%。7天后,仅在部分个体的血淋巴中检测到少量鳗弧菌。伴随着血淋巴液内鳗弧菌的减少,对虾的血淋巴细胞浓度发生变化。鳗弧菌注入对虾体内5 min后,血淋巴细胞浓度为(3.51±1.69)×107/mL,注射后2 h时达到最低值,为(2.39±1.76)×107/mL,然后逐步恢复,注射后48 h达到(3.97±1.60)×107/mL。对照组血淋巴细胞浓度起初略微降低,然后逐步增加,在2 h达到最大值后又逐渐降低;除注射后48 h外,对照组血淋巴细胞浓度始终高于实验组。鳗弧菌注射入斑节对虾体内后,不同的细胞种类呈现出不同的变化规律:在注射后2 h,对照组透明细胞占总细胞数的比例显著高于实验组,对照组半颗粒细胞的含量低于实验组。注射鳗弧菌后10 min,血淋巴中颗粒细胞的相对含量显著降低,此时实验组的颗粒细胞相对含量也低于对照组。而对于降解细胞细胞核的相对含量,实验组和对照组表现出了显著不同的变化趋势。在对照组中,24 h内其含量一直在降低,而后恢复;而对照组却在注射10 min时达到最高值,而后遵循对照组类似的变化。[中国水产科学,2006,13(1):28-32]     

饲料中不同蛋白含量对斑节对虾幼虾生长及消化酶的影响

选用鱼粉、豆粕和大豆浓缩蛋白为主要蛋白源,配置6个蛋白质水平梯度(36%、38%、40%、42%、44%、46%)的饲料,对斑节对虾幼虾(1.03±0.02 g)进行56 d养殖实验,研究饲料中不同蛋白含量对斑节对虾幼虾生长及消化酶活性的影响。结果表明,(1)摄食40%蛋白饲料的斑节对虾的增重率(417.35%)和特定生长率(2.93%·d-1)获得最大值并显著高于36%蛋白饲料组(P<0.05),但与其它各组无显著差异。饲料蛋白水平在38%时的饲料系数(1.88)最低且与44%和46%蛋白饲料组存在显著性差异(P<0.05)。(2)虾体蛋白含量在44%饲料蛋白组最高,且显著高于36%、38%和42%组(P<0.05)。(3)斑节对虾的蛋白质表观消化率在42%蛋白组最高,44%和46%蛋白饲料组的蛋白质效率显著低于36%～42%蛋白饲料组(P<0.05)。(4)随着蛋白水平的提高,斑节对虾的蛋白酶活性呈现由高到低波动的趋势;淀粉酶活性随着饲料蛋白含量的升高而呈下降趋势,胃淀粉酶活性在38%～42%蛋白饲料组显著高于36%蛋白饲料组(P<0.05),肠淀粉酶活性在42%蛋白饲料组显著高于其它各组(P<0.05)。通过饲料中蛋白质含量与增重率的回归分析,斑节对虾幼虾饲料蛋白的适宜含量为39.70%。     

斑节对虾增殖细胞核抗原基因克隆及表达分析

为研究增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)在斑节对虾(Penaeus monodon)卵巢发育中的功能,利用RACE技术克隆得到了斑节对虾PCNA基因(PmPCNA)的cDNA全长序列。该序列全长978 bp,包括135 bp的5′非编码区(5′UTR)、60 bp的3′UTR和编码260个氨基酸的783 bp的开放阅读框(ORF)。同源性分析结果表明,PmPCNA与其他物种具有很高的蛋白同源性。组织表达模式分析表明,PmPCNA在所监测的各组织中为组成性表达,其中在卵巢和脑中表达量较高;卵巢发育不同阶段基因的表达分析则表明PmPCNA在Ⅲ期表达量最高,是其他各期表达量的2倍;注射五羟色胺(5-hydroxy tryptamine,5-HT)后,PmPCNA在卵巢中表达量明显上升,其中在48 h表达量升高幅度最大,是对照组的5倍左右。利用原核表达技术获得了PCNA的体外重组蛋白,Western Blot分析证实该重组蛋白为PCNA蛋白。以上研究结果表明,PmPCNA蛋白是斑节对虾卵巢发育过程中的重要调节因子,本研究为进一步认识斑节对虾卵巢发育机制提供了基础资料。     

Integration of substrate in biofloc based system: Effects on growth performance,water quality and immune responses in black tiger shrimp,Penaeus monodon culture

To evaluate effect of substrate integration in biofloc based system, a 52‐day growth experiment was conducted using black tiger shrimp, Penaeus monodon juveniles (3.32 ± 0.07 g). The factorial design consisted of floc, F (with or without) as first factor and substrate (bamboo mat, B; nylon mesh, N; and without substrate) as second factor. This resulted six treatments; F + B, F + N, F, B, N and a control without biofloc and substrate. Shrimps were stocked at 110 nos. m^–3 in Fibre Reinforced Plastic (FRP) tanks and, rice flour was used as carbon source in biofloc based treatments. Incorporation of nylon mesh and bamboo mat in biofloc system trapped 31.3%–38.6% and 8.5%–13.5% total suspended solids respectively and reduced bottom solid deposition. Among the substrate based groups, significantly better development of biofilm with higher microbial population noticed in F + B compared with nylon mesh. Similarly, significantly higher final growth (p < 0.01) was recorded in F + B system followed by F + N while no significant difference in body weight recorded among floc, F or substrate based groups (B, N). Biofloc and substrate integration (F + B and F + N) resulted significantly (p < 0.01) lower feed conversion ratio compared to control and floc. Incoporation of bamboo substrate in biofloc, (F + B) improved shrimp immune responses through higher hemocyte counts and prophenoloxidase activity compared to other treatments. The study revealed that integration of substrate in the biofloc system improved growth performance, FCR and immune parameters in shrimp by trapping the suspended biofloc particles, better water quality parameters, enhanced biofilm growth and provision of quality natural food.     

Acrosome reaction in the sperm of the black tiger shrimp Penaeus monodon (Decapoda, Penaeidae)

Owing to the problem of male infertility in the domesticated shrimp Penaeus monodon, this study was conducted to reveal the morphological events of an acrosome reaction (AR) of sperm of this highly valuable species. The AR observed in an in vitro incubation of sperm with egg water (EW) and that during actual spawning was compared. Under transmission and scanning electron microscopy, sperm taken from the female thelycum was composed of a posterior main body, a central cap and an anterior single spike. Upon contact with EW, the sperm underwent two phases of AR: acrosomal exocytosis and spherical mass formation. The former was composed of a degeneration of the spike, swelling of the cap region and rupture of the acrosomal pouch. The latter began with polymerization of materials within the subacrosomal region and ended with re‐configuration of the subacrosomal region into an electron‐dense spherical mass. The AR of the sperm observed during spawning revealed similar morphological events, with degeneration of the spike upon contact with the vitelline envelope and formation of the spherical mass while penetrating into jelly material produced by protruding cortical rods. The results suggest the presence of AR inducers derived from the vitelline envelope and cortical rods of the egg. This study forms the basis for an evaluation of infertility regarding to AR in the domesticated P. monodon male.