Therapeutic effects of enrofloxacin in an experimental infection with a luminescent Vibrio harveyi in Artemia franciscana Kellog 1906

Although a wide range and number of chemotherapeutants have been developed and applied in aquaculture, there have been no studies on the absorption of antibiotics by crustacean larvae and, consequently, none has been reported to treat an established infection.

The aim of this study was to evaluate the therapeutic effects of enrofloxacin during an experimental infection with the strain PN9801 identified as Vibrio harveyi in Artemia franciscana.

Four treatments were used: (1) Nauplii experimentally infected with V. harveyi; (2) Nauplii enriched with enrofloxacin; (3) Nauplii enriched with enrofloxacin and, 4 h later, experimentally infected with V. harveyi; and (4) Nauplii experimentally infected with V. harveyi and 24 h later enriched with enrofloxacin. Survival rates were estimated after 48 h and they were as it follows: 29%, 99%, 88% and 85% for treatments 1, 2, 3 and 4, respectively.

This study is the first one to show some evidence that antibiotics can, in fact, withhold the course of a bacterial infection already established in Artemia and possibly in crustaceans.     

The effects of ozone and probiotics on the survival of black tiger shrimp (Penaeus monodon)

Total ozone production (TOP) from an ozonator, residual ozone concentration (ROC) in water, and the effects of ozone with or without probiotic supplemented feeds on bacterial growth, and shrimp (Penaeus monodon) survival were investigated. Minimal effective ROC to inhibit 3 log units of Vibrio harveyi D331 for 6 h and 2 log units of Bacillus S11 for 9 h was 0.38 g O₃/l of ROC from 5-min ozonation. Shrimp postlarvae exposed to 0.34–0.50 mg O₃/l ROC (8-h ozonation) caused loss of balance, immobility and destruction of gill lamellar epithelium. In vivo treatment of juvenile P. monodon reared on probiotic feed for 1 month revealed that 0.35 mg O₃/l ROC (30-min ozonation) effectively inhibited 3 log units of V. harveyi D331 for 24 h. At this ROC dosage, there was no effect either on shrimp or on intestinal probiotics (Bacillus S11). Shrimp survival from probiotic treatment, coupling with ozonation, increased significantly (P≤0.05) compared with controls.     

In vitro antimicrobial susceptibility of Photobacterium damselae subsp. piscicida to 15 different antimicrobial agents

The in vitro susceptibilities of 145 Photobacterium damselae subsp. piscicida strains, isolated from different geographical regions, against 15 major fish farming antimicrobial agents were assessed using a disc diffusion method. Ninety-nine percent of all isolates were sensitive to florfenicol and trimethoprime-sulfamethoxazole; 93% of the strains were resistant to erythromycin. Based on the antimicrobial susceptibility, the P. damselae subp. piscicida isolates could be distinguished from each other. While all American and European isolates were sensitive to kanamycin, 49% of the Japanese isolates were resistant to it. Moreover the results clearly indicated increasing resistance to a variety of antibiotics, particular in Japan, but also in some Mediterranean countries.     

4株长牡蛎(Crassostrea gigas)致病性哈维氏弧菌(Vibrio harveyi)鉴定及其毒力基因检测

哈维弧菌(Vibrio harveyi)是海水鱼养殖中一种重要的病原菌。该研究根据已知rbsB (核糖体结合蛋白)基因序列设计引物,扩增得到哈维弧菌354 (V.harveyi 354) rbsB基因的全序列(GenBank序列号MF797015),预测该序列可编码292个氨基酸,分子量为30.7 kD,理论等电点为5.05,亲水性系数为0.043,为疏水性蛋白。根据RbsB氨基酸构建的系统进化树可以发现哈维弧菌RbsB蛋白和欧文弧菌(V.owensii CAIM 1854)的关系最近。构建了pGEX-4t-1-rbsB重组质粒并转化至大肠埃希菌BL21 (DE3)中,得到的重组蛋白的相对分子量约59 kD,在37 ℃、IPTG浓度为0.6 mmol·L^–1诱导8 h时表达量最高。     

斑节对虾GRP94基因的克隆及其在不同应激条件下的表达与分析

细胞分裂周期蛋白42 (CDC42)是一类广泛表达的小三磷酸鸟苷 (GTP)酶,是Ras家族内Rho亚家族的成员之一,具有GTP酶活性。CDC42在细胞吞噬和炎症发生等先天免疫中发挥着不可替代的功能。该研究通过RACE技术克隆得到了斑节对虾(Penaeus monodon)细胞分裂周期蛋白42 (PmCDC42)基因cDNA全长。生物信息学分析结果表明PmCDC42全长2 233 bp,包括59 bp的5' 非编码区 (UTR)、1 598 bp的3'UTR和576 bp的开放阅读框 (ORF),编码191个氨基酸。通过实时定量PCR技术,研究了PmCDC42在斑节对虾各组织的表达模式,并研究了其在不同pH、盐度、重金属镉(Cd)和哈维弧菌 (Vibrio harveyi)胁迫下的表达情况。结果显示,PmCDC42基因在各组织中均有表达,在血淋巴中的表达量最高。在pH和Cd胁迫下PmCDC42表达量显著升高;在高盐胁迫下PmCDC42表达量上调,低盐胁迫下PmCDC42表达量变化不显著;在哈维弧菌的刺激下PmCDC42上调表达极显著。这表明PmCDC42在斑节对虾先天免疫中起重要作用。     

酵素菌生物有机鱼肥在水产养殖上的应用研究

为探讨自制环保酵素对水产病原微生物的抑制作用,该研究对其发酵菌株进行了分离。通过比浊法和平板划线法,确定自制环保酵素对水产病原微生物坎氏弧菌SP-1 (Vibrio campbellii SP-1)、哈维氏弧菌SP-1 (V. harveyi SP-1)、轮虫弧菌SP-1 (V. rotiferianus SP-1) 和嗜水气单胞菌245 (Aeromonas hydrophila) 的最低抑制浓度分别是体积分数是体积分数为11.5%、11%、6.2%和11%,最小杀菌浓度分别是体积分数为16%、16%、16%和12%。攻毒保护试验表明,自制环保酵素无菌上清液可使嗜水气单胞菌对斑马鱼 (Danio rerio) 的致死率降低 (90.5±6.49) %。pH、温度、蛋白酶敏感性试验显示自制环保酵素的主要抗菌活性物质为酸性物质,对加热和蛋白酶处理稳定。进一步从环保酵素中分离主要发酵菌株,通过ITS序列分析鉴定为库德毕赤酵母 (Pichia kudriavzevii)。研究结果可为自制环保酵素在水产养殖中对抗细菌性疾病的应用提供理论基础。

     

大黄鱼内脏白点病的病原分析与鉴定

为实现大黄鱼 (Larimichthys crocea) 内脏白点病的生物防控,推进水产养殖用药减量,从健康大黄鱼肠道中筛选出对其内脏白点病病原菌——杀香鱼假单胞菌 (Pseudomonas plecoglossicid) 有拮抗作用的益生菌。采用琼脂扩散法筛选菌株,通过生理生化特征以及分子生物学分析,对菌株进行鉴定,并评价其溶血性、药敏性、安全性、产酶能力及抗菌广谱性。从健康大黄鱼肠道中初步分离出37株潜在益生菌,通过拮抗试验筛选出3株具有拮抗效果的菌株,分别命名为P1-17、P2-33和P3-11。通过生理生化特性及16S rRNA 测序分析,菌株P1-17和P2-33鉴定为贝莱斯芽孢杆菌 (Bacillus velezensis),菌株P3-11鉴定为粪肠球菌 (Enterococcus faecalis);溶血性试验和药敏试验结果表明3株菌均无显著溶血圈,且所含耐药因子较少,不具备潜在致病性;人工回感试验证实,3株拮抗菌对健康大黄鱼无致病性;抗菌谱测定结果表明,2株芽孢杆菌对溶藻弧菌 (Vibrio alginolyticus)、哈维氏弧菌 (V. harveyi)、美人鱼发光杆菌 (Photobacterium damselae) 等多种水产病原菌具有拮抗效果,同时2株芽孢杆菌具有产淀粉酶和蛋白酶的能力;而该株粪肠球菌只对杀香鱼假单胞菌有拮抗作用。研究结果可为后续大黄鱼肠道益生菌的筛选和应用提供科学依据。     

水产健康养殖理念与发展对策

为了给哈维氏弧菌(Vibrio harveyi)疾病防治新手段的开发提供基础实验数据,筛选了协同抗哈维氏弧菌的乌梅(Fructus mume)水提液与抗生素复合物。首先绘制药物浓度对数对抑菌圈直径标准曲线,采用全组合法筛选与乌梅水提液具有协同抗菌活性的抗生素,然后采用中心组合设计法(CCD)结合响应面法(RSM)分析乌梅水提液与筛出抗生素复合后两两药物间的交互作用,并确定最佳复合配比。全组合法筛出的四环素、土霉素、金霉素和强力霉素分别与乌梅水提液具有协同抗菌作用,RSM分析显示将这4种药物复合后,乌梅水提液与土霉素对哈维氏弧菌存在显著协同抗菌作用;土霉素分别与金霉素和强力霉素存在交互抑制作用;乌梅水提液、土霉素、金霉素和强力霉素最佳复合配比为45∶0.12∶0.00∶0.00 (mg·mL^–1)。模型预测最优复合物对哈维氏弧菌的抑菌圈直径为18.54 mm,琼脂扩散法平行3次测得该复合物的抑菌圈直径为(19.03±0.07) mm,相对误差为2.6%,表明该方法结果可靠,为考察药物间的交互作用提供了新思路。     

斑节对虾CDK2基因全长cDNA克隆及表达分析

凋亡抑制因子1 (tumor protein p53 regulated inhibitor of apoptosis 1, TRIAP1)可以通过抑制细胞凋亡参与生物体内应激反应。为探究TRIAP1基因及调控其表达的miRNAs在斑节对虾 (Penaeus monodon) 应对哈氏弧菌(Vibrio harveyi)刺激时的表达调控情况,该研究通过RACE技术获得了斑节对虾TRIAP1基因(PmTRIAP1)cDNA全长序列,并利用实时荧光定量PCR (qRT-PCR)技术,对PmTRIAP1在斑节对虾不同组织中的表达分布模式,以及哈氏弧菌刺激下PmTRIAP1与相关miRNAs的表达关联情况进行了探究。结果显示,PmTRIAP1 cDNA全长2 522 bp,包括11 bp的5'非编码区(UTR)、2 289 bp的3'UTR和222 bp的开放阅读框(ORF),共编码73个氨基酸。PmTRIAP1基因在各组织中均有表达,其中在血细胞中表达量最高;哈氏弧菌刺激下,PmTRIAP1表达量显著降低,而Pm-miR-145-3p和Pm-miR-454-3p表达量显著升高,表明PmTRIAP1和Pm-miR-145-3p、Pm-miR-454-3p的表达量呈负相关关系;双荧光素酶报告实验结果显示,Pm-miR-145-3p和Pm-miR-454-3p可通过结合PmTRIAP1 3'UTR降低荧光素酶活性,表明Pm-miR-145-3p和Pm-miR-454-3p均可靶向调控PmTRIAP1的表达。

     

Immunological and pathological status of gilthead sea bream (Sparus aurata L.) under different long-term feeding regimes

The possible influence of the feeding regime (FR) on the immune system and pathological status of gilthead sea bream was studied. Two growth trials were performed starting at different seasons (trial 1=March; trial 2=June) under controlled experimental conditions. In both trials, FR-1 groups received a restricted amount of food, whereas FR-2 groups were fed to visual satiety. The pathology study included parasitological and bacteriological examination, and the immunological traits analysed were respiratory burst activity of head kidney leucocytes, serum lysozyme and alternative pathway complement activity (ACH₅₀). The immunological status of gilthead sea bream not only was not impaired by the restricted feeding regime, but also seemed to be enhanced in some aspects, as the respiratory burst of FR-1 fish of trial 2 was significantly higher. No differences in the bacterial isolates were detected between the two feeding regimes, and Vibrio harveyi was the most prevalent species in both cases, especially in warm months. Also, fish under FR-1 regimes had significantly lower mortality, lower prevalence of infection of all the parasites except Cryptosporidium molnari, and less histopathological alterations in liver and intestine than those under FR-2 regimes.     

非洲斑节对虾健康养殖技术

该研究通过RACE (Rapid-amplification of cDNA ends) 法克隆并获得斑节对虾 (Penaeus monodon) 甲壳动物雌性激素 (Crustacean female sex hormone, CFSH) 基因PmCFSH的开放阅读框(ORF)及3'非编码区(UTR),预测编码214 氨基酸(aa)的蛋白,其含有信号肽和1个与免疫应答相关的白介素17E (IL-17E) 结构域。qRT-PCR结果显示,PmCFSH基因在各个组织中均有表达,其中在腹神经组织中的表达量最高;从受精卵时期到仔虾期的表达量呈逐渐上升趋势,其中在仔虾期表达量最高;在卵巢发育Ⅱ—Ⅳ期,随卵巢发育成熟表达量逐渐升高;革兰氏阴性菌和阳性菌均能上调PmCFSH基因的表达量,其中鳃组织PmCFSH在哈维氏弧菌 (Vibrio harveyi) 感染后第3小时和金黄色葡萄球菌 (Staphylococcus aureus) 刺激后第12小时达到表达高峰;而肝胰腺PmCFSH在哈维氏弧菌感染后第3小时和金黄色葡萄球菌刺激后第48小时达到表达高峰。研究表明,PmCFSH基因不仅参与幼体发育以及性腺成熟的调控,对细菌的应激也具有免疫应答响应,体现了激素的“多功能性”现象。

     

贝类幼虫面盘纤毛脱落病的起因及防治

为研究文蛤育苗过程中幼虫病害及其主要致病菌,实验通过构建细菌16S rDNA克隆文库、病原菌分离纯化、人工感染和药敏实验等方法对育苗场发病的浮游期文蛤壳顶幼虫样品进行了系统分析。结果显示,发病的文蛤壳顶幼虫细菌群落多样性低,地中海弧菌占比高达75%以上,推断其可能为引发此次幼虫发病的主要致病菌。从发病幼虫的匀浆组织中分离获得该优势菌株,测序及系统发育鉴定为地中海弧菌。人工感染实验确定了其致病性,菌株US2-01在1.0 × 10⁶ CFU/mL 的菌液浓度下浸泡感染文蛤浮游幼虫,96 h累计死亡率为84%。药敏实验表明,地中海弧菌菌株US2-01在12种抗生素的测试中对常用的青霉素、氨苄西林、红霉素等抗生素具有一定的耐药性,对四环素和多西环素中度敏感,对头孢他啶、庆大霉素、卡那霉素等其余7种抗生素呈现高度敏感。本研究首次报道了地中海弧菌为文蛤浮游期幼虫致病的一种潜在病原,研究结果可为文蛤幼虫疾病研究及贝类苗种培育过程中的病害防控提供科学依据。     

Bacterial diversity of tilapia (Oreochromis niloticus) cultured in brackish water in Saudi Arabia

The bacterial flora occurring in brackish pond water, sediment, gills and intestine of healthy tilapia cultured in Saudi Arabia were estimated both quantitatively and qualitatively, and the isolates were identified to genus or species level. Total viable count of bacteria ranged from 1.4±1.5×10³ to 8.6±2.7×10³ cfu ml⁻¹; 1.2±3.1×10⁶ to 7.3±1.1×10⁷ cfu g⁻¹; 8.7±1.9×10⁵ to 2.1±0.9×10⁶ cfu g⁻¹; and 2.8±2.4×10⁷ to 1.0±1.6×10⁸ cfu g⁻¹ in the pond water, sediment, gills and intestine of brackish water tilapia, respectively. In total, 19 bacterial species were identified. The bacteria were predominantly Gram-negative rods (87%). Pond water and sediment bacteria influenced the bacterial composition of gills and intestine of tilapia. In contrast to gill bacteria, more diversification was observed in intestinal bacteria. The predominant (prevalence >10%) bacterial species were Vibrio parahaemolyticus, Vibrio carchariae, Vibrio alginolyticus, Chryseomonas sp., Vibrio vulnificus, and Streptococcus sp. in all the populations with the exception of the sediment population where Streptococcus sp. was replaced by Shewanella putrefaciens. Vibrio spp. (58% of the total isolates) dominated the total bacterial population.     

Effect of water temperature on the immune response of white shrimp Litopenaeus vannamei to Vibrio alginolyticus

White shrimp Litopenaeus vannamei held in 25‰ seawater at 27 °C or 28 °C were injected with TSB-grown Vibrio alginolyticus at 1 × 10⁴ colony-forming units (cfu) shrimp^− 1 or 1 × 10⁵ cfu shrimp^− 1, and then cultivated onward at water temperatures varying from 20 to 34 °C. Over 24–144 h, mortality of V. alginolyticus-injected shrimp held at 34 °C or 32 °C was significantly higher than that of shrimp held at lower temperatures. In a separate experiment, shrimp held in 25‰ seawater at 28 °C and then cultured onward at 20 to 32 °C were examined for immune parameters at 24–96 h. THC, phenoloxidase activity, respiratory burst, and SOD activity decreased significantly at 24 h after transfer to 32 °C. Shrimp held in 25‰ seawater at 27 °C and then cultured onward at 20 to 34 °C showed a significant reduction in phagocytic activity and clearance efficiency for V. alginolyticus at 24 h after transfer to 34 °C. It was concluded that transfer of shrimp from 27 or 28 °C to higher temperatures (32 and 34 °C) reduced their immune capability and decreased resistance to V. alginolyticus infection.     

Rearing of flounder (Platichthys flesus) juveniles in semiextensive systems

A low-technology rearing system was implemented for rearing juvenile flounder for stock enhancement in a Danish fjord, the Limfjord. Each year during 1996–2002, between 13,000 and 153,000 juveniles were reared from the yolk-sac stage until metamorphosis in outdoor ponds relying on phyto- and zooplankton blooms as their main food source. In contrast to other similar systems, the blooms in this system are closely monitored and, to a certain extent, regulated. The zooplankton blooms consisted mainly of calanoid copepods, dominated by the species Temora longicornis and Centropages hamatus. Most juveniles produced (>99.5%) were normally pigmented with average yearly survival rates from hatch to metamorphosis varying from 7±9% to 48±18%, lowest in the first years of production.     

Biofilm development within a larval rearing tank of the tropical rock lobster, Panulirus ornatus

The role of bacterial biofilms in disease processes is becoming increasingly recognised in both clinical and environmental settings. Biofilm development within a rearing tank of the tropical rock lobster Panulirus ornatus was studied to evaluate if the biofilm is a reservoir for potentially pathogenic bacteria that cause mass larval mortalities. Within a 5000 L larval rearing tank, fiberglass microscope slides were systematically distributed during a standard rearing attempt to assess biofilm development. Culture-based counts for two media types, TCBS and Marine Agar (MA), demonstrated increased bacterial densities until days 11 and 13 respectively. For both media types, a drop in the plate counts was followed by a subsequent increase towards the end of the experiment. Scanning electron microscopy (SEM) confirmed that cell densities decreased between days 13 and 17, most likely due to sloughing of the biofilm into the water column. SEM images revealed distinct changes in dominant morphologies reflecting a succession of bacterial populations. A dynamic succession of microbial species during biofilm development was also demonstrated using denaturing gradient gel electrophoresis (DGGE) profiling of bacterial 16S rRNA genes in combination with statistical ordination analysis. Prominent changes in the DGGE profiles coincided with the decrease in bacterial numbers observed by SEM and plating on MA between days 13 and 17. Fluorescence in situ hybridization (FISH) identified -Proteobacteria as being numerically abundant in the biofilm. This was supported by results from DGGE analysis, which retrieved only sequences affiliated with - and γ-Proteobacteria. DGGE bands affiliated with Vibrio became dominant towards the end of the larval run (days 21 to 24). A Vibrio harveyi strain isolated from the biofilm late in the larval rearing trial (day 24) demonstrated increased larval mortality in small scale phyllosoma survival studies. The detection of Vibrionaceae at the end of the larval trial coincided with mass phyllosoma mortality and show that the biofilm is a reservoir for potentially pathogenic bacteria.     

水产品养殖销售环节致病性弧菌污染状况和病原特征分析

该研究采用双层平板涂布法和划线法,从不同地区的海洋环境样品中分离纯化光合细菌,以副溶血弧菌 (Vibrio parahemolyticus)、创伤弧菌 (V. vulnificus)、鳗弧菌 (V. anguillarum) 为指示菌,采用牛津杯法测定海洋光合细菌菌株的抑菌作用,采用靛酚蓝分光光度法和盐酸萘乙二胺分光光度法测定不同菌株对氨氮 (NH₄ ⁺-N) 和亚硝态氮 (NO₂ ⁻-N) 的降解作用,筛选出具有抗弧菌并高效降解NH₄ ⁺-N和NO₂ ⁻-N复合功能的优良菌株。结果显示,从30个海水、海泥等样品中分离得到3株光合细菌,分离自连云港车牛山岛海水样品的菌株P-3,对3种弧菌均具有较强的抑制作用,其中对鳗弧菌的作用最强,抑菌圈直径为5.3 mm。3株光合细菌均具有一定的降解NH₄ ⁺-N和NO₂ ⁻-N作用,菌株P-3的降解作用最强,在含有50 mg·L⁻¹ NH₄ ⁺-N和NO₂ ⁻-N的培养基中培养4 d,降解率分别为89.68%和94.98%。经形态学观察、生理生化试验和16S rDNA序列分析,确定P-3为沼泽红假单胞菌 (Rhodopseudomonas palustris)。     

Isolation,identification and pathogenicity of Vibrio harveyi,the causal agent of skin ulcer disease in juvenile hybrid groupers Epinephelus fuscoguttatus × Epinephelus lanceolatus

The hybrid grouper, Epinephelus fuscoguttatus (♀) × Epinephelus lanceolatus (♂), is a newly bred cultivated marine fish species of high economic value. However, a skin ulcer disease with high mortality has occurred, and the responsible pathogen remains unknown. In this study, we summarized the epidemic status and external signs of this disease. We screened potential pathogens and finally isolated one bacterial strain ML01 from affected fish. We subjected healthy juvenile hybrid groupers to bacterial challenge tests with the isolate by immersion, immersion after dermal abrasion and intraperitoneal injection, respectively. Within 14 days post‐infection, the isolate ML01 caused mass mortality of juveniles infected via immersion after dermal abrasion or intraperitoneal injection. Diseased juveniles displayed obvious signs of skin ulcers. The median lethal dose of ML01 by intraperitoneal injection was 1.10 × 10⁵ colony‐forming units. ML01 was identified as Vibrio harveyi by bacterial morphology, analytical profile index identification, 16S rDNA sequencing and multilocus sequence analysis. Antibiotic susceptibility tests showed that ML01 was sensitive to ceftriaxone, doxycycline and minocycline. The results of this study suggest that V. harveyi is the causal agent of skin ulcer disease in juvenile hybrid groupers, thus providing a basis for effective control and prevention of this disease.     

Furunculosis in Atlantic salmon (Salmo salar L.) is not readily controllable by bacteriophage therapy

The potential of bacteriophage therapy to control bacterial disease in farmed fish was tested using, as an example, furunculosis of Atlantic salmon, caused by Aeromonas salmonicida subsp. salmonicida.

In vivo testing with Atlantic salmon and rainbow trout (Oncorhynchus mykiss Walbaum) showed no adverse effects, with bacteriophage generally cleared within 96 h of administration by either intraperitoneal (i.p.) injection or oral in-feed.

Juvenile Atlantic salmon were administered a combination of bacteriophage O, R and B (1.9 × 10⁸ pfu fish^− 1) by i.p. injection, after they had been challenged with A. salmonicida subsp. salmonicida 78027, also by i.p. injection. The fish that were injected with bacteriophage immediately after challenge died at a significantly slower rate then those that were either not treated with bacteriophage, or treated 24 h post-challenge. However, the end result (100% mortality) was not affected.

In further experiments the effects of oral (1.88 × 10⁵ pfu g^− 1 fish^− 1 daily for 30 days), bath (1.04 × 10⁵ ml^− 1 daily for 30 days) and i.p. (6.25 × 10⁷ pfu fish^− 1) phage treatment to control furunculosis in experimentally infected Atlantic salmon were compared with antibiotherapy (treatment with 10 mg kg^− 1 bw^− 1 day^− 1 oxolinic acid for 10 days), using an indirect cohabitation challenge. No protection was offered by any of the bacteriophage treatments, compared to the positive challenge group, although significant protection was offered by the oxolinic acid treatment. Analysis of samples taken from the trials demonstrated that bacteriophage were correctly administered to the fish and, on occasion, were isolated from fish that had succumbed to furunculosis. It was also shown that bacteriophage resistant A. salmonicida subsp. salmonicida isolates could be recovered from mortalities in all the treatment groups.

The results suggest that, although there were no safety problems associated with the approach, furunculosis in Atlantic salmon is not readily controllable by application of bacteriophage.     

Influence of different dietary amino acid patterns on growth and body composition of juvenile Japanese flounder, Paralichthys olivaceus

A feeding trial using five semi-purified diets (50% crude protein) was conducted to investigate the effects of different dietary amino acid patterns on growth and body composition of juvenile Japanese flounder. The control diet contained casein and gelatin as intact protein sources and four other diets contained 30% casein–gelatin (2:1, w/w) and 20% crystalline amino acids (CAA). CAA were added to the diets to simulate the amino acid pattern found in red sea bream egg protein (REP), Japanese flounder larvae whole body protein (FLP), Japanese flounder juvenile whole body protein (FJP), and brown fish meal protein (BFP), respectively. The test diets were fed to triplicate groups of juveniles (2.75±0.05 g) twice a day for 40 days to evaluate weight gain, survival, feed conversion efficiency (FCE), protein efficiency ratio (PER), and apparent protein utilization (APU). The apparent retention of total dietary amino acids in the whole body and A/E ratios of the whole body were also evaluated. The highest weight gain was observed in fish fed the diet containing the dietary amino acid pattern of BFP followed by fish fed the control, FJP, FLP and the REP diets. Percent survival, FCE, PER and APU were also significantly (P<0.05) affected by the amino acid pattern in the diets, indicating the highest value in fish fed the BFP diet. Except for a few amino acids, the amino acid composition of the whole body did not show marked differences with different dietary amino acid pattern. Results suggest that BFP could be more suitable as a reference amino acid pattern in the diet of juvenile Japanese flounder compared to the amino acid pattern of FLP, FJP or REP.