南美白对虾池塘养殖水质智能控制系统设计与开发

南美白对虾池塘养殖对于水体质量有着较为严苛的要求,残饵及生物排泄物的增多会大量消耗水体溶解氧,进而打破水体动态平衡,导致NH₃-N、NO₂-N等毒害成分升高,引起南美白对虾病害甚至导致死亡,造成养殖户经济损失并且阻碍了南美白对虾养殖产业的健康发展。针对上述问题,笔者研究并开发一种能够实时监测养殖池塘水质,并能根据南美白对虾生长习性智能控制增氧机、水泵等养殖设备的南美白对虾池塘养殖水质智能控制系统。系统具有控制智能、运行可靠、集成度高等特点,在养殖生产试验中取得了良好效果,保证了南美白对虾养殖过程的安全,有效降低了南美白对虾池塘养殖风险。     

Impact of farm management on expression of early mortality syndrome/acute hepatopancreatic necrosis disease (EMS/AHPND) on penaeid shrimp farms in Thailand

Asian shrimp farming industry has experienced massive production losses due to a disease caused by toxins of Vibrio bacteria, known as early mortality syndrome/acute hepatopancreatic necrosis disease (EMS/AHPND) for the last 5 years. The disease can cause up to 100% cumulative pond mortality within a week. The objective of this study was to identify factors associated with AHPND occurrence on shrimp farms. A case–control study was carried out on shrimp farms in four provinces of Thailand. Factors related to farm characteristics, farm management, pond and water preparation, feed management, post‐larvae (PL) shrimp and stock management were evaluated. Multivariable logistic regression analysis identified factors affecting AHPND occurrence at the pond level. Chlorine treatment, reservoir availability, use of predator fish in the water preparation, culture of multiple shrimp species in one farm and increased PL stocking density contributed to an increased risk of AHPND infection, while delayed first day of feeding, polyculture and water ageing were likely to promote outbreak protection. Additionally, the source of PL was found to be associated with AHPND occurrence in shrimp ponds, which requires further study at the hatchery level. Identification of these factors will facilitate the development of effective control strategies for AHPND on shrimp farms.     

斑节对虾Chitinase-2基因的克隆及其在蜕皮和幼体发育过程中的表达分析

研究以斑节对虾(Penaeus monodon)转录组获得的几丁质酶基因(Pm Chi)片段,运用RACE技术克隆了PmChi基因c DNA全长,命名为PmChi-2。PmChi-2基因c DNA全长2 050 bp,其中5'-非编码区(5'-UTR)144 bp、3'-UTR 319 bp和开放阅读框(ORF)1 587 bp,编码528个氨基酸。生物信息学分析显示,PmChi-2与其他甲壳动物Chi-2的相似性为78%~97%。实时荧光定量PCR结果显示,PmChi-2在蜕皮前期(D期)表皮中表达水平最高,在胃、鳃、腹神经节和眼柄中表达水平依次降低,在其他组织(肝胰腺、肠、肌肉、心脏)中几乎不表达。不同蜕皮阶段,PmChi-2在鳃、胃和表皮3种组织中的表达变化模式基本一致,均在蜕皮期(E期)表达量最低,而最高表达量在鳃中出现在蜕皮后期(A期),在胃和表皮中为D期。幼体不同发育阶段分析揭示PmChi-2 mRNA在幼体发育阶段的无节幼体到糠虾幼体第二期表达量维持在一个低水平,在糠虾幼体第三期PmChi-2 mRNA表达水平显著升高,在仔虾期又显著下降,推测PmChi-2 mRNA可能与斑节对虾幼体发育密切相关。研究结果表明PmChi-2基因可能在斑节对虾蜕皮以及幼体的变态发育中发挥重要作用,为深入研究斑节对虾几丁质酶发育调控提供了重要信息。     

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Toll9受体基因免疫功能的研究与表达分析

Toll受体蛋白(Toll receptors)是一类重要的模式识别受体,在无脊椎动物先天性免疫系统中发挥重要作用。文章对斑节对虾(Penaeus monodon)新型Toll9受体基因(Pm Toll9)进行了研究:以人源胚胎肾细胞(HEK293T)成功构建体外细胞免疫模型,通过免疫印迹方法证实Pm Toll9重组真核蛋白可在HEK293T中成功表达。双荧光素酶报告系统检测发现在200 ng转染浓度处Pm Toll9对NF-κB报告基因的激活效果显著。q RT-PCR数据证明PmToll9成功激活HEK293T细胞Toll-like receptor(TLR)信号通路,促进通路下游髓样分化因子(My D88)、肿瘤坏死因子(TNF-α)和白介素(IL-10)的上调表达,同时酶联免疫吸附测定结果证明Pm Toll9可促进TNF-α蛋白表达水平显著上调。斑节对虾体内细菌刺激实验结果显示无乳链球菌(Streptococcus agalactiae)可激活Pm Toll9在肝胰腺、肠、淋巴和鳃中的表达,哈维氏弧菌(Vibrio harveyi)可显著抑制Pm Toll9在肝胰腺中的表达。提示无乳链球菌可通过Pm Toll9激活Toll信号通路,引起机体免疫防御反应,而哈维氏弧菌对此过程具有一定的抑制作用。     

Effects of adding sucrose on Penaeus monodon (Fabricius, 1798) growth performance and water quality in a biofloc system

A 60‐day indoor growth trial was conducted to study the effects of biofloc on the growth performance of a Penaeus monodon (Fabricius, 1798), water quality and biological indicators including biofloc volume, chlorophyll‐a, heterotrophic bacteria and Bacillus quantity. Two concentrations of sucrose (0 and 75%) were added daily to the P. monodon culture systems (2.94 ± 0.11 g), which were conducted indoors in fibre‐glass tanks (500 L). Results showed that the final body weight and weight gain of the adding 75% sucrose group were significantly higher (P < 0.05) than that of the control, as well as significantly (P < 0.05) improved specific growth rates and survival rates, and reduced feed coefficient. Adding 75% sucrose promoted heterotrophic bacteria, Bacillus and phytoplankton reproduction, and significantly (P < 0.05) reduced the concentration of ammonia‐N (NH₄‐N), nitrite‐N (NO₂‐N) and nitrate‐N (NO₃‐N). The changes of water quality indicators in the two groups showed the similar trend at the end of the experiment, and the ammonia‐N, nitrite‐N, nitrate‐N and phosphate‐P concentrations in the 75% sucrose group were significantly (P < 0.05) lower than those of the control group, Chlorophyll‐a concentrations peaked at 389.12 μg/L in the biofloc sucrose group at 18:00 h, and heterotrophic bacteria peaked 8 h after sucrose was added. The addition of sucrose also reduced the pH of the water. Our research showed that adding sucrose promoted biofloc formation and shortened the formation time; increased the number of heterotrophic bacteria and algae which might play a role in improving water quality by assimilating ammonia‐N and other harmful substances in the water; supplemented food for P monodon growth; and reduced the feed coefficient.     

Effect of vitamin E on spermatophore regeneration and quality of pond‐reared,black tiger shrimp (Penaeus monodon)

The objective of the present work was to evaluate the effects of different levels of vitamin E in feed on the spermatophore regeneration and quality of male Penaeus monodon. The experiment was carried out with the four following treatments: the basal diet no added vitamin E, the diet added 200, 600 and 1,000 mg/kg respectively. Spermatophore regeneration and quality were evaluated by spermatophore weight, sperm count and spermatophore absence rates, which male P. monodon were extruded spermatophore for feeding 20 and 40 days. In the experiment, the weight of the twice regenerated spermatophore of the males added to the vitamin E group was higher than that of the untreated control group. The weight of the first regenerated spermatophore with the addition of 1,000 mg/kg group was the highest and significantly higher than the control group (p < .05), but there was no significant difference among the three groups with different levels of vitamin E. The weight of the second regenerated spermatophore with the addition of 600 mg/kg group was the highest, followed by 1,000 mg/kg group, both of which were higher than the control group and the addition of 200 mg/kg group. Within the same group, the regeneration spermatophore weight showed overall upward trend as the feeding time, twice regenerate experiment spermatophore weight with added to the vitamin E groups were significantly higher than the initial value (p < .05), but three spermatophore weight of male shrimp at the control group had no significant difference. The sperm quantity and the percentage of normal sperm of the twice regenerated spermatophore of the males with added to the vitamin E group was higher than that of the untreated control group, and those of the addition of 200 mg/kg group was significantly higher than that of control group (p < .05). The total number of sperm and the percentage of living sperm of male shrimp in the experimental group decreased with the increase of vitamin E in the feed. Within the same group, the total number of sperm and the percentage of living sperm of male shrimp with added to the vitamin E groups showed overall upward trend as the feeding time and were significantly higher than the initial value (p < .05), but the control group was slightly down and had no significant difference. Comprehensive sperm weight, sperm quantity and living sperm percentage of three indicators, that adding 200 mg/kg of vitamin E in feed could effectively promote the spermatophore regeneration in the male P. monodon and improve the sperm quantity. The experimental results provide a scientific basis for the breeding of P. monodon.     

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.     

Dietary administration of Bacillus (B. licheniformis and B. subtilis) and isomaltooligosaccharide influences the intestinal microflora,immunological parameters and resistance against Vibrio alginolyticus in shrimp,Penaeus japonicus (Decapoda: Penaeidae)

The experiment was conducted to investigate the effects of oral administration of probiotics (Bacillus licheniformis and Bacillus subtilis) in combination with prebiotic isomaltooligosaccharide (IMO) on the intestinal microflora and immunological parameters of Penaeus japonicus and its resistence against Vibrio alginolyticus. A basal diet was supplemented with 0% probiotics and prebiotic (control), 10⁸ colony forming unit (CFU) g^?1B. licheniformis and 0.2% IMO (T1), 10⁸ CFU g^?1B. subtilis and 0.2% IMO (T2), 10⁸ CFU g^?1B. licheniformis in combination with 10⁸ CFU g^?1B. subtilis and 0.2% IMO (T3). The results showed that the total bacterial counts significantly increased (P<0.05) and Vibrio counts significantly decreased (P<0.05) in the intestine of shrimp with supplementation of dietary synbiotics. Shrimp fed the diet with both Bacillus probiotics and IMO (T3) produced significantly higher immune parameters (phenoloxidase activity, lysozyme activity and nitric oxide synthase activity and superoxide dismutase activity) than the control group (P<0.05). Significant lower cumulative mortality after challenge with V. alginolyticus was also observed in shrimp fed diet with both Bacillus probiotics and IMO (T3) than that in the control group (P<0.05). The results indicated that the oral administration of probiotics (B. licheniformis and B. subtilis) and prebiotic IMO has positive effects on bacterial flora in P. japonicus gut, and can activate non‐specific immunity of shrimp.