Intestinal microbiota of healthy and unhealthy Atlantic salmon Salmo salar L. in a recirculating aquaculture system

The present study sampled the intestinal content of healthy and unhealthy Atlantic salmon( Salmo salar L.), the ambient water of unhealthy fish, and the biofilter material in the recirculating aquaculture system(RAS) to understand differences in the intestinal microbiota. The V4–V5 regions of the prokaryotic 16 S r RNA genes in the samples were analyzed by Mi Seq high-throughput sequencing. The fish were adults with no differences in body length or weight. Representative members of the intestinal microbiota were identified. The intestinal microbiota of the healthy fishincluded Proteobacteria(44.33%), Actinobacteria(17.89%), Bacteroidetes(15.25%), and Firmicutes(9.11%), among which the families Micrococcaceae and Oxalobacteraceae and genera S phingomonas, Streptomyces, Pedobacter, Janthinobacterium, Burkholderia, and Balneimonas were most abundant. Proteobacteria(70.46%), Bacteroidetes(7.59%), and Firmicutes(7.55%) dominated the microbiota of unhealthy fish, and Chloroflexi(2.71%), and Aliivibrio and Vibrio as well as genera in the family Aeromonadaceae were most strongly represented. Overall, the intestinal hindgut microbiota differed between healthy and unhealthy fish. This study off ers a useful tool for monitoring the health status of fish and for screening the utility of probiotics by studying the intestinal microbiota.

Establishment and Characterization of a Fin Cell Line Derived from the Atlantic Salmon Salmo salar and Its Application to Fish Virology Study

Atlantic salmon(Salmo salar)is an important economic fish that is seriously threatened by various viruses.A cell line designated as ASF derived from the caudal fin tissue of Atlantic salmon was established and characterized in this study.ASF cells grew well in Dulbecco’s modified Eagle’s medium(DMEM)containing 20%fetal bovine serum at 20℃.DNA sequencing and comparative analysis of the cytochrome B gene verified that the ASF cell line originated from Atlantic salmon.Chromosome analysis indicated that the modal chromosome number of ASF cells was 58.Viral susceptibility test showed that ASF cells were susceptive to two important fish viruses,viral hemorrhagic septicemia virus(VHSV)and red-spotted grouper nervous necrosis virus(RGNNV).Viral replication in ASF cells was further confirmed by qRT-PCR and transmission electron microscopy.Moreover,VHSV and RGNNV infections could induce the cellular responses in ASF cells,as indicated by the differential expression of cellular antiviral response-related genes,interferon-1 and Mx-1.In conclusion,the newly established ASF cell line can be applied as an in vitro tool in fish virology and immunity studies.

复合益生菌制剂对大西洋鲑(Salmo salar)生长、消化酶和非特异性免疫指标的影响

选取体质健康、均重为(180±28)g的大西洋鲑(Salmo salar)420尾,随机分成4组(对照组和3个不同菌剂水平的实验组),每组3个重复,每个重复35尾。对照组投喂基础饲料,实验组分别投喂添加1%、3%和5%芽孢杆菌和酵母菌复合益生菌制剂(BSCP)的基础饲料42 d。结果发现,大西洋鲑投喂1%–5%的BSCP后生长性能得到明显改善,增重率显著升高(P<0.05),饵料系数和死亡率显著降低(P<0.05);饲料蛋白质的表观消化率呈升高的趋势,5%组蛋白质消化率具有显著性差异(P<0.05),但其脂肪表观消化率显著降低(P<0.05);实验组肠道和肝脏蛋白酶和脂肪酶活性有不同程度的升高,但其中5%组肝脏脂肪酶活性显著降低(P<0.05);实验组肝脏免疫指标表现出不同程度的改善,但其血清免疫指标无明显变化,仅中高剂量组SOD活性显著降低(P<0.05)。结果表明,BSCP可以显著提高大西洋鲑的生长性能,提高部分消化酶活性和蛋白质的消化率,并在一定程度上促进大西洋鲑的非特异性免疫能力,其饲料中适宜添加水平为3%。

改性粘土絮凝法对大西洋鲑(Salmo salar)存活及生理生化的影响初探

本文以平均体重为(388.00±41.11)g的大西洋鲑(Salmo salar)为实验对象,探究了改性粘土絮凝过程对养殖水体水质以及大西洋鲑存活、组织形态和氧化应激反应的影响。在本实验条件下,添加改性粘土的实验组水质得到一定程度的改善,其中磷酸盐、铵盐、亚硝酸盐浓度相较于对照组有明显降低的趋势(P<0.05)。添加远高于现场可有效去除赤潮生物用量(4-10g/m^2)的改性粘土并没有导致大西洋鲑出现死亡现象,显微观察结果显示,实验组与对照组中大西洋鲑鱼鳃、消化道组织特征无明显差别。对大西洋鲑鳃、肝脏、消化道的丙二醛(MDA)含量、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性测定结果表明,改性粘土未对大西洋鲑造成显著的氧化胁迫(P>0.05)。本研究结果说明,在远高于现场有效用量条件下,改性粘土未对大西洋鲑产生不良影响,该研究结果将为改性粘土治理有害藻华技术在国内外养殖海域的应用提供科学依据。

Effects of spectral composition,photoperiod and light intensity on the gonadal development of Atlantic salmon Salmo salar in recirculating aquaculture systems(RAS)

Artificial lighting regimes have been successfully used to inhibit sexual maturity of Atlantic salmon in confinement.However,when these operations are applied in commercial recirculating aquaculture systems(RAS) using standard lighting technology,sexual maturation is not suppressed.In this study,an L_9(3~3) orthogonal design was used to determine the effects of three factors(spectral composition,photoperiod,and light intensity) on the gonadal development of Atlantic salmon in RAS.We demonstrated that the photoperiod at the tested levels had a much greater effect on the gonadosomatic index and female Fulton condition factor than spectral composition and light intensity.The photoperiod had a significant effect on the secretion of sex steroids and melatonin(P<0.05),and a short photoperiod delayed sex steroid and melatonin level increases.The three test factors had no significant effects on the survival rate,specific growth rate,relative weight gain,and male Fulton condition factor(P>0.05).The optimum lighting levels in female and male Atlantic salmon were LD 8:16,455 nm(or 625 nm),8.60 W/m^2;and LD 8:16,8.60 W/m^2,455 nm respectively.These conditions not only delayed gonadal development,but also had no negative effects on Atlantic salmon growth in RAS.These results demonstrate that a combination of spectral composition,photoperiod and light intensity is effective at delaying the gonadal development of both male and female salmon in RAS.

Effects of feeding and stocking density on digestion of cultured Atlantic salmon Salmo salar L.

The combined effects of feeding rate(0.8%,1.0%,and 1.2%initial body weight/day),feeding frequency(two,three,and four times/day) and stocking density(10,15,and 20 kg/m3) in recirculating aquaculture systems(RAS) on growth performance,digestion and waste generation of Atlantic salmon(Salmo salar L.) were investigated in an 8-week orthogonal experiment(L9(3)3) with a constant daily water renewal at 7.50%of total volume.No mortality occurred during the experimental period.Feed conversion ratio(FCR) varied from 0.90 to 1.13 and specific growth rate(SGR) ranged from 0.48%to 0.69%/day.SGR,thermal growth coefficient(TGC) and FCR were not significantly(P>0.05) affected by the three factors,while net protein utilization(NPU) was significantly(P<0.05) affected.Apparent digestibility coefficients(ADC) of dry matter in the present study were in the range 66.12%-73.55%.ADC in protein,lipid and energy were statistically different among all treatments and in the range of 90.07%-93.67%,81.54%-89.15%,and67.55%-71.87%,respectively.The proportion of mean total ammonia nitrogen excreted ranged from 1.37%to 1.64%of feed nitrogen at steady state,and the concentration of nitrogenous and phosphorus compounds were differently correlated to the three factors.The results will provide valuable reference data for culture management decisions in the Atlantic salmon farming industry.

Evaluation of Feed and Feeding Regime on Growth Performance, Flesh Quality and Fecal Viscosity of Atlantic Salmon(Salmo salar L.) in Recirculating Aquaculture Systems

The effects of different feeds and feeding regimes on growth performance, flesh quality and fecal viscosity of Atlantic salmon(Salmo salar L.) in recirculating aquaculture systems(RAS) were investigated. Fish(initial body weight of 1677 g ± 157 g) were fed with four commercial feeds(Nosan salmon-NS, Aller gold-AG, Skretting salmon-SS and Han ye-HY) in two feeding regimes(80% and 100% satiation) for 78 d. The results showed that salmon specific growth ratio(SGR) and weight gain ratio(WGR) were significantly affected by feed type and feeding regime(P < 0.05). Feed conversion ratio(FCR) varied between 0.93 and 3.40, which was significantly affected by feed type(P < 0.05), and slightly improved with increased satiation degree. The activities of digestive enzymes including protease, lipase and amylase were also significantly affected by feed type and feeding regime(P < 0.05), increasing with satiation degree. Flesh qualities for vitamin E, hydroxyproline(HYP), liquid loss and muscle p H among all groups showed significant differences(P < 0.05), ranging from 26.67 to 29.67, while no obvious difference was found in flesh color. Fecal viscosity for different treatments showed no significant difference, though improvement was found in 100% satiation group. From present experiment, it was concluded that both feed type and feeding regime can affect the important quality attributes of Atlantic salmon.

Suliman and Waliya (a Salar folk story)

SULIMAN was a young Salar hunter who possessed a magic bow and knife. At the age of 25, he was eager to find a good wife. One day, at the foot of a mountain, he saved a skylark from the mouth of a water snake. When the skylark song thanks to Suliman, he saw a pretty girl in the

The Salar Women

The Beijing Zhaotai Real Estate Development Corporation headed by Mu Cunyu has a capital of 360 million yuan.

Hanhaqimei,a Salar Woman

The Salar nationality,with apopulation of 87,697,mainly settles in Xunhua Salar Nationality Autonomous County in eastern Qinghai Province.The Salar have their own spoken language,but no written script.Most of the Salar use Mandarin Chinese.According to legend and historians,their ancestors movde to Xunhua from Middle Asia in the 13th century.The Salar living by the Yellow River engage in agriculture and animal husbandry as their Primary means of support. The Salar believe in Islam and are also greatly influenced by confucian culture developed by the Han nationality.

基于生物质谱的蛋白质组学分析大西洋鲑中生物标志性肽段及过敏原特异性肽段

基于蛋白质组学自下而上研究策略,采用纳升液相色谱-四极杆-静电场轨道阱高分辨质谱联用法(Nano-UHPLC-Q/Exactive-HRMS)和超高效液相色谱-串联质谱法(UHPLC-MS/MS)筛选、鉴定大西洋鲑中生物标志性肽段和过敏原β-小清蛋白特异性肽段,并提出了UHPLC-MS/MS测定其含量的方法。称取5 g处理好的样品,加入10 mL正己烷除脂,离心,于沉淀中加入5 mL磷酸盐缓冲溶液(提取剂),涡旋,于60℃水浴加热30 min,再用20 mL冷丙酮富集、纯化,得到的蛋白提取液经胰蛋白酶酶解后,采用Nano-UHPLC-Q/Exactive-HRMS初步筛选大西洋鲑生物标志性肽段与过敏原β-小清蛋白特异性肽段,用UHPLC-MS/MS对初筛的肽段进行多反应监测(MRM)模式验证。在酶解后的溶液中加入内标,采用UHPLC-MS/MS测定,内标法定量。结果显示:筛选出3条可用于定量的大西洋鲑生物标志性肽段VAVNVEETK、VAPLSEDFK、LTEIQSLER和1条过敏原β-小清蛋白特异性肽段TFFHTIGFASK;VAVNVEETK、VAPLSEDFK、LTEIQSLER标准曲线的线性范围在5.00μmol·L^(-1)以内,TFFHTIGFASK标准曲线的线性范围在10.00μmol·L^(-1)以内,检出限(3S/N)为0.32~1.55μg·g^(-1);空白样品中4条肽段的加标回收率为90.2%~105%,测定值的相对标准偏差(n=6)为1.5%~4.6%;方法用于测定大西洋鲑贮存过程中VAVNVEETK、VAPLSEDFK、LTEIQSLER、TFFHTIGFASK的含量,LTEIQSLER和TFFHTIGFASK含量没有发生明显变化,而VAVNVEETK和VAPLSEDFK含量随着贮存天数的延长逐渐降低,其中VAPLSEDFK适用于大西洋鲑新鲜度评价。

我国撒拉族民间医药调研分析及活态保护策略研究

目的:对我国撒拉族民间医药的调研数据进行对比分析和总结归纳,为撒拉族民间医药的活态保护提供思路和数据支撑,进一步提出该民族民间医药活态保护策略。方法:采用轮次实地走访青海省循化撒拉族自治县及毗邻撒拉族主要生活区的方法,补充和完善调研数据。最终通过文献研究法、报表统计法、座谈讨论法、综合归纳法、回访核实法,对调研数据进行分析及总结。结果:完成青海省循化撒拉族自治县及毗邻撒拉族主要生活区的84个撒拉族行政村60岁以上撒拉族老人和民间医务工作者的调研,共获得了2095份第一手资料,共整理出撒拉族民间医药相关名词术语600条。其中,人体组织器官等名称66条、疾病相关的词条信息54条、诊断方法200条、医用器具22条、饮食起居120条、药物信息108条、特色外治与康养保健30条。结论:本次调研为撒拉族民间医药传承、发展及活态保护提供了基础数据。该民族对其自身医药文化的保护意识淡泊,加之当地无专门的民族医药发展研究机构,制约了该民族医药的发展。希望当地有关部门采取设立撒拉族民间医药社团和发展协会等措施,对本民族民间医药进行活态保护。

程度显赫:撒拉语及亲属语言形容词的程度等级序列

阿尔泰语系语言的传统语法研究普遍认为形容词具有通过形态标记表达的级语法范畴,若从语义及形态句法的角度分析,这类形态标记与印欧语中的平比级、比较级以及最高级等典型级范畴成员存在显著差异,应该界定为形容词的程度范畴。文章从黏着标记-^(0)rAQ入手,提出了级范畴与程度范畴的十条界定标准,将撒拉语及亲属语言形容词的程度范畴分为弱化程度、强化程度以及主观评价程度三大类。相较于印欧语中的级范畴,以撒拉语为代表的阿尔泰语系语言可选用黏着形态、重叠以及词汇—句法手段对形容词属性的不同程度进行精细化的区分,编码形式十分丰富,一些语言中的弱化程度标记甚至衍生出表达委婉语气的扩展功能,语法化程度较高。整体来看,程度范畴是撒拉语等阿尔泰语系语言形容词的显赫范畴,其内部又具有主观性与客观性的区别,主观性的程度主要通过不同的黏着形态标记以及复杂多变的重叠手段表达,而客观性程度主要通过程度副词来进行区分,客观程度内部呈现出最大量>极大量>偏大量>过量>偏小量的程度渐变等级序列。由于语言接触等因素的影响,撒拉语程度范畴黏着形态标记的使用频率逐渐降低,具有一些较为凸显的个性特点。

贝藻混养对大西洋鲑养殖废水的生物滤除

养殖废水的综合利用与无公害化处理排放是实现水产养殖业健康可持续发展的重要保障。作者采用太平洋牡蛎(Crassostrea gigas)及龙须菜(Gracilaria lemaneiformis)混养和单养的方式处理大西洋鲑工业化循环水养殖系统排放的废水,设置贝类组、藻类组、贝藻组3组处理,探讨了贝藻混养方式和贝类、藻类单养对废水中主要水质因子(N、P营养盐、化学需氧量(COD)和总悬浮颗粒物(TSS))的处理效率,实验周期为30 d。结果表明,牡蛎和龙须菜混养的方式处理养殖废水效果较好,其对氮、磷营养盐、COD及TSS的去除效率分别为:总氨态氮41.67%±8.82%、硝酸盐氮33.96%±0.34%、总磷7.18%±0.03%、COD 78.87%±1.82%和TSS 70.50%±1.65%,而亚硝酸盐氮出现一定的积累。综合分析,牡蛎和龙须菜混养的方式处理养殖废水的效率优于牡蛎和龙须菜的单独处理。

脂肪和蛋白质营养对封闭循环水养殖大西洋鲑生长和肌肉品质的效应

为探讨饲料脂肪和蛋白质营养对工业化养殖大西洋鲑（SalmonsalarL．）生长性能和肌肉品质的影响，采用3×2双因素试验设计（3脂肪水平：18％、21％、24％，即F18、F21、F24；2蛋白质水平：38％、48％．即P38、P48），形成6种试验处理膨化颗粒配合饲料，每处理3重复，通过在工业化封闭循环海水养殖条件下，选用初质量（650．0±45．50）g大西洋鲑720尾，进行动物饲喂试验、肌肉脂肪酸、氨基酸测试等，试验期56d。结果表明：（1）中脂肪水平（F21）和高蛋白水平（P48）饲料的生长性能较佳。增质量率随脂肪水平提高先增加后降低，中高脂肪组比低脂肪组极显著提高43．5％（P〈0．01）；高蛋白组极显著提高44．99％（P〈0．01）；中脂肪高蛋白组合（P48F21）效应最佳；（2）脂肪水平对肌肉氨基酸含量影响不显著。蛋白水平与肌肉9种必需、4种风味氨基酸含量呈正相关，高蛋白组显著提高3．60％～17．00％（P〈0．05／0．01）：（3）脂肪水平与肌肉不饱和脂肪酸含量呈正相关，而蛋白水平与其呈负相关，高脂肪（P24）和低蛋白（P38）有利于提高肌肉不饱和脂肪酸含量。高脂肪纽肌肉Zoo-3PUFA、∑HuFA、DHA、EPA含量比低脂肪组分别极显著或显著提高15．74％、26．60％、15．41％、7．67％（P〈0．01／0．05）；低蛋白组极显著提高肌肉DHA含量11．91％（P〈0．01）、EPA10．02％（P〈0．05）；低蛋白中高脂肪组合fP38F21、P38F24）效应佳。本试验研究证明，工业化工业化封闭循环水养殖大西洋鲑对脂肪营养需求有一定程度降低，本试验条件下中高脂肪与中低蛋白组合既利于提高生长性能，又增加肌肉ω-3HUFA和主要氨基酸沉积，为工业化养殖大西洋鲑的肉质改善型配合饲料研制提供了重要依据。

基于蛋白质/肽组学的三文鱼物种溯源

不同三文鱼在外观和物种亲缘关系上都十分相近,本研究旨在建立一种大西洋鲑鱼、大马哈鱼和虹鳟鱼的物种溯源方法。基于质谱鸟枪法蛋白质组学,在95%置信水平上共鉴定得到912个蛋白质。经过对不同三文鱼特征蛋白质相应肽段的检测特异性和物种特异性筛选分析,得到潜在的物种特征肽段生物标志物。通过多反应监测的靶向肽组学,进一步验证物种特征肽段生物标志物,得到了大西洋鲑鱼10条物种特征肽段、大马哈鱼2条物种特征肽段和虹鳟鱼3条物种特征肽段,并对市售的商品三文鱼产品进行了方法应用。结果显示,该方法操作方便、准确可靠、特异性好、适用性强,可在常规分析实验室对商品三文鱼产品进行有效的鉴别分析。

大西洋鲑杀鲑气单胞菌的分离鉴定

从皮肤溃烂的大西洋鲑(Salmon salar)肌肉、肝、肾分离到一致病性的菌株(AB080226),经人工感染实验证实其为该病的病原菌,研究了该菌的形态、生理生化特征并对其16SrDNA序列进行了在线Classifer和Blast分析。结果显示:该菌为革兰氏阴性杆菌,葡萄糖氧化发酵阳性,氧化酶检测阳性,0%NaCl生长;能利用甘露醇,不具运动性,能发酵麦芽糖,葡萄糖产酸阳性,精氨酸双水解酶阳性,苯丙氨酸脱氨酶阴性;该菌属于气单胞菌属的细菌,在系统发育树上与杀鲑气单胞菌形成一个簇群,同源性高达99.9%以上。综合形态学、生理生化特征及16SrDNA序列鉴定其为杀鲑气单胞菌(Aeromonas salmonicida)。

超声成像技术在大西洋鲑早期性别及发育期鉴别的应用研究

采用KX-5100便携式B超机（7.5MHz高频线阵探头）对273尾1龄或2龄大西洋鲑（Salmo salar）的性别及性腺发育阶段进行了检测,同时对检测的大西洋鲑进行了解剖和取样,利用组织学观察法验证了超声成像技术对大西洋鲑雌雄性别和发育阶段的鉴别结果。研究结果显示：利用超声成像技术鉴别雌、雄大西洋鲑的平均准确率为86%,且鉴别雌鱼的准确率（93%）较雄鱼（81%）高。超声成像技术鉴别Ⅱ期＋Ⅲ期、Ⅳ期、Ⅴ期卵巢的准确率分别为87%、92%和100%,鉴别Ⅱ期、Ⅲ期、Ⅳ期、Ⅴ期精巢的准确率分别为64%、71%、86%和84%。总体看来,超声成像技术鉴别早期性腺（Ⅱ期~Ⅲ期）发育的准确率较低,仅为75%,而对晚期性腺（Ⅳ期~Ⅴ期）发育鉴别的准确率较高,达到89%。

大西洋鲑在封闭循环水养殖模式下的生长和性腺发育研究

本研究探讨了在封闭循环水养殖模式下大西洋鲑(Salmo salar)的生长发育模式,对封闭循环水系统养殖的大西洋鲑重要生长数量性状进行生物学测量,并跟踪性腺发育状况。本研究自2012年1月通过封闭循环水系统进行大西洋鲑养殖,经过周年养殖,体质量平均达到1194.42g,体长平均达到417.00mm,体高平均达到100.58mm,体宽平均达到53.52mm,少数个体在养殖10个月后性腺发育较快,性腺质量达到9.76g,性腺指数达到1.31%。本研究的研究结果为建立大西洋鲑标准化循环水养殖系统积累资料。