一例日本囊对虾暴发性死亡的病原分析

为了阐明山东省潍坊市一对虾养殖场发生日本囊对虾暴发性死亡的原因,采用分子生物学检测方法,对发病对虾进行了白斑综合征病毒(white spot syndrome virus,WSSV)、桃拉综合征病毒(Taura syndrome virus,TSV)、黄头病毒(yellow head virus,YHV)、传染性皮下及造血组织坏死病毒(infectious hypodermal and hematopoietic necrosis virus,IHHNV)、传染性肌肉坏死病毒(infectious myonecrosis virus,IMNV)、偷死野田村病毒(covert mortality nodavirus,CMNV)及急性肝胰腺坏死病(acute hepatopancreatic necrosis disease,AHPND)7种病原的检测,且对发病对虾进行了常规组织病理学观察。同时采用16S r DNA细菌鉴定方法及浸泡回接感染实验对分离自发病对虾体内的可疑病原菌进行了分子鉴定及毒力测试。结果显示,发病对虾样品核酸检测呈现WSSV强阳性,IHHNV和CMNV为弱阳性,其他4种病原为阴性。组织病理学观察发现,在对虾的胃、鳃等上皮组织中存在WSSV包涵体,头部肌肉纤维出现离散。对分离编号为2901、2902、2903的3株优势可疑病原菌鉴定结果显示,3株菌分别与印度格里蒙菌、交替单胞菌及溶藻弧菌相似,相似度分别为99%、99%及100%。攻毒结果显示,3株可疑病原菌的LC50分别为9.8×107、1.1×108与2.3×108 CFU/m L,各细菌毒力均较弱,非导致对虾出现暴发性死亡的病原。研究表明,导致本次日本囊对虾暴发性死亡的病因与混合感染病原WSSV、IHHNV、CMNV有关,其中WSSV感染是造成日本囊对虾暴发性死亡的主因,研究结果可为解析当前养殖日本囊对虾疾病暴发及其成因提供参考。     

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。     

Evaluation of growth,feed utilization efficiency and immune parameters in tiger shrimp (Penaeus monodon) fed diets supplemented with or diet fermented with gut bacterium Bacillus sp. DDKRC1. isolated from gut of Asian seabass (Lates calcarifer)

An experiment was conducted to assess the effect of supplementation of cellulolytic gut bacterium with plant protein‐based diet on growth, digestibility and digestive enzyme status of tiger shrimp, Penaeus monodon. Bacillus sp. DDKRC1. (JN641289), a potential cellulolytic bacterium, isolated from the gut of Asian seabass was used as feed supplement in this experiment. The shrimps of group I, II and III were fed with control diet (without bacterial supplement, D1), diet supplemented with live Bacillus sp. DDKRC1. at 2.94 × 10⁷ cfu per 100 g feed (D2) and diet fermented (48 hr) with same bacteria (D3) respectively. At the end of the 42 days experiment, shrimps of group II showed significantly better (p < .01) growth and protein efficiency ratio (PER) and lower (p < .01) feed conversion ratio (FCR) as compared to group I and III. Dry matter, cellulose, hemicellulose and lipid digestibilities were significantly (p < .01) higher in group II than other two groups. Cellulase, amylase and protease activities in the gastrointestinal (GI) tract were significantly (p < .01) higher in group II as compared to other two groups. Total heterotrophic count, amylolytic, cellulolytic and proteolytic bacterial counts in gut were significantly (p < .01) higher in group II as compared to other two groups. Haematological analysis indicated better (p < .05) immune response in groups II and III than control group. Diets D2 and D3 showed better effect on growth, digestibility, FCR, survival and immune response of P. monodon as compared to D1.     

遮阴对珠子参生长发育及光合生理特性的影响

为探明遮阴对珠子参生长发育和光合生理特性的影响,对盆栽珠子参分别采取全光照、30%遮阴和50%遮阴3种处理。在珠子参生长90天后,考察各处理下珠子参植物学性状、生理指标和光合特性。结果表明,与全光照相比,30%遮阴处理下珠子参株高、主茎粗、果实数、根茎直径、叶绿素含量、SOD和POD活性增加,地下根茎节间长度和叶MDA含量减小。遮阴处理后珠子参净光合速率(Pn)、气孔导度(Gs)和蒸腾速率(Tr)低于全光照处理,胞间CO_2浓度(C_i)则高于全光照处理。遮阴有效地缓解了珠子参的"午休"并降低了光抑制,其中,30%遮阴处理效果最显著,为珠子参的生长提供了较为适宜的环境。     

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。     

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.     

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.     

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.