黄颡鱼养殖高产新技术

黄颡鱼属底层鱼类,肉质鲜嫩少刺,商品价值高。规模养殖对池塘建设、鱼种投放、饲料、水质等都有一定的要求,文章根据养殖实践,就黄颡鱼养殖基础设施准备、鱼苗鱼种放养规格、放养个体大小、饲料选择、科学投喂、病害防治等技术进行了介绍。     

黄颡鱼Sox9基因的分离及分析

Sox9基因是号陛别分化形成有关的重要基因之一。以黄颡鱼（Pelteobagrus fulvidraco）为研究材料，使用RT-PCR在脑、精巢和卵巢分离和克隆到了黄颡鱼Sox9基因，序列比对分析表明，脑和精巢Sox9基因序列一致，称Sox9al（GenBank accession No．AY684328），卵巢的则与其有差异称Sox9a2（GenBank accession No．AY684329）。Sox9al与Sox9a2的核苷酸相似性为77％，氨基酸的相似性为84％，可以认定是由两个基因表达的。和其它动物Sox9氨基酸序列的同源性分析表明：黄颡鱼Soxga1与其它动物Sox9的相似性在8l％，89％之间，平均在86％左右；黄颡鱼Sox9a2的相似性在76％～86％，平均在83％左右。系统树分析揭示了Sox9为单起源，遗传距离远近也与传统分类结果一致。使用RT-PCR对Sox9a1与Sox9a2在雌雄黄颡鱼前脑和性腺组织的表达检测结果显示，Sox9a1在雌雄鱼的前脑和性腺均有表达，但Sox9a2只在卵巢表达，说明该基因可能与雌性性别形成及卵巢发育具有调控作用。     

黄颡鱼的含肉率及肌肉营养价值研究

采用常规方法测得广西桂江黄颡鱼的含肉率为 83.88% ;每 1 0 0 g肌肉 (干样 )中含粗蛋白85.96g,粗脂肪 5.0 4 g,粗灰分 5.57g,氨基酸总量为 70 .2 1 g,必需氨基酸含量 2 7.39g,含钙 39.58mg,磷 1 87mg,铜 0 .33mg,锌 5.33mg,铁 4 .75mg,锰 3.0 3mg,硒 74 7.0 6μg。综合分析结果认为 ,广西桂江的黄颡鱼是营养价值较高的优质鱼类     

六价铬对黄颡鱼仔鱼和稚鱼的急性毒性效应研究

利用静水式鱼类急性毒性测试法,分别以黄颡鱼（Pelteobagrus fulvidraco）仔鱼和稚鱼为受试对象,通过六价铬的急性毒性试验,获得24、48、72、96 h相应的半数致死浓度LC50值,比较了六价铬对黄颡鱼仔鱼和稚鱼的致毒敏感性,计算了六价铬对黄颡鱼的安全质量浓度并对六价铬对于黄颡鱼急性毒性等级进行了评价。旨在通过开展我国本土水生生物的急性毒性效应的研究,获得相应的毒理学数据,为推导符合我国生态分区特点的水质基准提供基础信息。结果表明,在水温为（23±2）℃条件下,六价铬对黄颡鱼仔鱼24、48、72、96 h的半数致死浓度LC50分别为28.32、21.99、17.70、15.79 mg.L-1,而对黄颡鱼稚鱼24、48、72、96 h的LC50分别为138.5、88.36、68.55、57.98 mg.L^-1,说明黄颡鱼仔鱼对于六价铬毒性的敏感性明显高于稚鱼。六价铬对黄颡鱼仔鱼和稚鱼的安全浓度分别为1.579、5.798 mg.L^-1,依据急性毒性结果和鱼类急性毒性分级标准,六价铬对于黄颡鱼毒性等级属于中等毒性。     

黄颡鱼源维氏气单胞菌的分离、鉴定及特性分析

为确定导致江西南昌地区黄颡鱼腹水病暴发的病原，进行了病鱼的临床解剖观察、病原菌分离鉴定和回归感染、病原菌株药敏及毒力基因检测等系统性试验和分析。结果显示，患病鱼体鳃、肝脏、肾脏以及肠道等组织均发生不同程度的病变；由病鱼体内分离获得一株优势菌PL1，经过16S rDNA序列比对和系统进化树分析，鉴定为维氏气单胞菌（Aeromonas veronii）。回归感染结果表明，菌株PL1能够引起黄颡鱼与自然患病相同的典型腹水病病症，具有较强的致病性；药敏试验结果显示，菌株PL1对30种抗菌药物中多粘菌素、复方新诺明、呋喃唑酮等12种药物敏感，对氧氟沙星、红霉素和哌拉西林3种药物中度敏感，而对诺氟沙星、环丙沙星、万古霉素等15种抗生素具有耐药性，呈现出多重耐药性。毒力基因检测结果显示，菌株PL1携带有aerA、act、gcaT、ser、fla、ascV等6种毒力基因，表明菌株PL1具有较强的毒力。本研究为黄颡鱼腹水病的防治提供了科学指导，同时为深入研究患病机制提供了基础数据。     

黄颡鱼抗菌肽hepcidin基因的克隆和表达分析

用RT-PCR方法,从黄颡鱼（Pelteobagrus fulvidraco）肝脏中克隆获得了hepcidin cDNA序列,并对其进行序列测定,在此基础上分析了hepcidin基因在不同组织中的表达差异和不同细菌感染后对其表达的影响。结果表明,黄颡鱼hepcidin的cDNA片段（GenBank登陆号EU257703）长为282bp,编码93个氨基酸,由信号肽(23个氨基酸)、前肽（45个氨基酸）和成熟肽（25个氨基酸）组成,成熟肽含有8个保守的半胱氨酸（cys）残基,可形成4个链内二硫键。与已报道的其他鲶形目鱼类hepcidin核苷酸序列的同源性为80-82%,推导的氨基酸序列的同源性为69-71%。RT-PCR半定量分析显示黄颡鱼hepcidin基因在不同组织中普遍表达,其中肝脏表达水平最高,头肾、肠道、肌肉、脑、胃、鳃、心脏、卵巢表达次之,皮肤、脾脏、肾脏表达较低。感染嗜水单胞菌和腊样芽孢杆菌的黄颡鱼,肝脏hepcidin mRNA水平显著升高。     

褐藻糖胶对黄颡鱼幼鱼中脂肪酸的影响

为研究褐藻糖胶作为饲料添加剂对黄颡鱼幼鱼脂肪酸的影响,选取大小、体重相近的黄颡鱼幼鱼,随机分为对照组和试验组,对照组饲喂不添加褐藻糖胶的基础饲料,试验组饲喂添加0.1%褐藻糖胶的饲料,采用气相色谱质谱联用技术(GC-MS)分析不同喂养时间(1、2、4、8周)下试验组和对照组黄颡鱼幼鱼脂肪酸含量的变化。结果表明,黄颡鱼幼鱼体内共检测出15个总脂肪酸(TFA)和17个游离脂肪酸(FFA)。方差分析结果表明,黄颡鱼幼鱼TFA中C18:1 (n-9)、C20:4(n-5)和C22:6 (n-3)具有交互作用,而大部分FFA间都存在交互作用。喂养时间对黄颡鱼幼鱼脂肪酸有显著影响(P<0.05),随着喂养时间的延长,游离的单不饱和脂肪酸(MUFA)含量呈先减少后增加趋势,而总MUFA含量呈先增加后减少趋势;黄颡鱼幼鱼游离脂肪酸和总脂肪酸中饱和脂肪酸(SFA)均呈先增加后减少趋势,而多不饱和脂肪酸(PUFA)呈先下降后升高趋势。0.1%褐藻糖胶对黄颡鱼幼鱼TFA中C18:1 (n-9)、C20:4(n-5)和C22:6 (n-3)含量影响显著(P<0.05),对FFA组分含量的影响均显著(P<0.05),试验组黄颡鱼幼鱼SFA含量高于对照组,MUFA含量低于对照组,PUFA在喂养第2周时高于对照组,喂养第8周时低于对照组。此外,黄颡鱼幼鱼C20:3(n-3)/C22:6(n-3)的比值小于0.4,且喂养0.1%褐藻糖胶第8周时,黄颡鱼幼鱼C22:6(n-3)和C20:4(n-5)含量明显高于对照组,说明褐藻糖胶有益于黄颡鱼产卵和孵化,有助于黄颡鱼幼鱼的发育生长。本研究结果为黄颡鱼的健康养殖提供了基础数据。     

池塘80：20无公害黄颡鱼养殖技术

黄颡鱼,又叫黄腊丁,该鱼为温水性鱼类,属于杂食性底栖鱼类,喜集群,栖息于水体底层,在弱光条件下摄食和活动.适应性较强,可在各类水域中养殖.是目前较具发展潜力的淡水养殖品种,其肉质嫩,少刺无鳞,营养丰富,近几年市场非常畅销.已在网箱、池塘等水域中成功进行放养,并成为我县品种结构调整的主要名优鱼类之一.     

筛选杂交鲤亲子鉴定的微卫星标记(英文)

微卫星的亲子鉴定技术既能减少分池饲养带来的遗传性状的差异又能减少其所占用饲养池的数量和管理强度,在水产养殖和品种选育中得到了广泛的应用。本研究应用21对微卫星标记对鲤(Cyprinus carpio)杂交家系(49-同胞家系:德国镜鲤(C.carpio var.mirror)♀×荷包红鲤(C.carpio var.wuynanensis)♂;24-同胞家系:荷包红鲤♀×德国镜鲤♂)进行遗传多样性及遗传特性的评价,并检测影响微卫星标记鉴定准确性的几个特性。研究结果标明,(1)筛选出的13对标记:在49-同胞家系中,平均有效等位基因数(K)、期望杂合度(He)、观测杂合度(Ho)和多态信息含量(PIC)分别为0.505、0.648、0.791和0.5889;在24-同胞家系中,K、He、Ho和PIC分别为0.548、0.670、0.819和0.6138。(2)筛选出的13对标记:在49-同胞家系和24-同胞家系中的联合排除率(CPE)分别达到0.999997978和0.999999583。(3)在两个杂交家系中,4个核心位点(MFW29,HLJ392,HLJ044,HLJ855)的联合排除率均高于99%。(4)三种情况下(一个亲本已知,NE-1P;另一个亲本已知,NE-2P;双亲都已知,NE-PP;)的联合非父排除率在49全胞家系中分别为1.34E-02、3.91E-04和2.02E-06;在24全胞家系中分别为1.34E-02、3.91E-04和2.02E-06。(5)模拟分析显示(置信度为99%),当使用11个或更多标记时,在49全胞家系中亲本与子代的配对率为100%;当使用9个或更多标记时,在24全胞家系中亲本与子代的配对率大于98%。(6)UPGMA聚类图表明:在混养条件下,筛选出的13对标记可以明显地将子代与无亲缘关系的家系区分开。本研究筛选出13对微卫星标记完全适用于荷包红鲤与德国镜鲤杂交家系的鉴定,为品种选育提供有效的鉴定工具。     

微卫星标记在鱼类遗传及育种研究中的应用

微卫星标记在鱼类遗传多样性、种群遗传结构分析、基因组作图等方面的研究中发挥着较大的作用,并逐渐应用到鱼类分类、亲本鉴定及资源保护与育种等工作中。     

Isolation and Characterization of Phenol-Degrading Psychrotolerant Yeasts

Water, Air, & Soil Pollution - To investigate the differential contribution of extracellular polymeric substances (EPS) to polycyclic aromatic hydrocarbon (PAH) degradation by fungi and...     

Isolation and Characterization of Methanethiol-Producing Bacteria from Agricultural Soils

Methanethiol(MT) produced from terrestrial soils can have important effects on atmospheric chemistry and ecosystem-level processes, and it originates mainly from the metabolism of sulfur-containing amino acids by microorganisms.Methanethiol producing bacteria(MPB) were enriched and isolated from agricultural soils in a modified basal medium containing methionine(Met) as the sole carbon source.The isolates were identified as Bacillus sp.WH-R1, WH-R2, and WH-R3; Arthrobacter sp.SZLB-W3; and Delftia sp.CHZG-R4 based on cell morphology, physiological and biochemical characteristics, and 16 S r RNA sequence analysis.Delftia sp.CHZG-R4 was identified as a novel strain producing MT using Met as a precursor, and it had the most active MT-producing potential, with the production of MT being 21.8 μg at 30℃ and pH 7.0.Optimal MT production was observed at 35℃ and pH6.0, with 51.3% of sulfur content in Met being converted into MT.Under these conditions, MT production changed according to the supply of both carbon and nitrogen sources.The addition of 2 g L~(-1) each of starch, sucrose, urea, and potassium nitrate promoted MT production by more than 10%, whereas addition of 2 g L~(-1) each of ammonia sulfate and peptone decreased MT production by16% and 87%, respectively.This is the first study to report MT production by Delftia sp.CHZG-R4, providing useful information for the microbial mechanism of MT production from agricultural soils.Our findings also contribute to improving our knowledge of the function of Delftia sp.CHZG-R4.     

尼古丁降解菌L1的分离鉴定与降解特性分析

通过对分离于烟草叶片的154株具有降解尼古丁细菌菌株筛选，获得能高效降解尼古丁的L1菌株。通过形态观察，16S rDNA序列分析和生理生化测定将其鉴定为Bacillus simplex。菌落生长密度测定和高压液相色谱分析表明， L1菌株最适生长尼古丁浓度为1g/L，随着菌落密度增加，尼古丁降解效率逐渐升高，36 h最高降解率达到75.0%。而低尼古丁含量不利于L1菌株的生长，过高尼古丁含量对L1菌株的生长和降解效率具有反馈抑制作用。菌株L1降解尼古丁过程中无色素产生，说明其尼古丁代谢途径有别于节杆菌。     

Isolation and Characterization of Cadmium- and Arsenic-Absorbing Bacteria for Bioremediation

The contamination of hazardous metal(loid) is one of the serious environmental and human health risks. This study isolated a total of 40 cadmium (Cd)- and arsenic (As)-resistant bacterial isolates from coastal sediments by pour plate technique using tryptic soy agar supplemented with Cd or As (50 mg l^?1) for use as metal(loid) bioremediation agents. Out of 40, 4 isolates, RCd3, RCd6, RAs7, and RAs10, showed a relatively higher growth rate in Cd- or As-supplemented culture media which were selected for further study. The selected isolates showed a high minimum inhibitory concentration (60–400 mg l^?1 for Cd and 400–2200 mg l^?1 for As), which demonstrated their remarkable Cd and As resistance capabilities. The metal(loid) removal efficiencies (0.032–0.268 μg Cd h^?1 mg^?1 and 0.0003–0.0172 μg As h^?1 mg^?1 [wet weight cell]) of selected isolates indicated their greater magnitude in absorbing Cd compared to As from water. Phylogenetic analysis of the 16S rDNA sequences revealed that isolates RCd3, RCd6, RAs7, and RAs10 were closely related to Acinetobacter brisouii, Pseudomonas abietaniphila, Exiguobacterium aestuarii, and Planococcus rifietoensis, respectively. Because of high Cd and As resistance and removal efficiency, the selected isolates can survive in a high metal(loid)-contaminated environment and could be a potential tool for bioremediation of high metal(loid)-contaminated effluents to protect the aquatic environment.     

鸡胚原始生殖细胞的分离、培养及鉴定

采用Ficoll密度梯度离心法分离第19期(孵化68～72 h)艾维茵鸡(Gallus domesticus)生殖嵴原始生殖细胞(PGC)，通过体细胞-PGC共培养进行了原代和传代培养，对PGC进行了碱性磷酸酶(AKP)和过碘酸雪夫氏反应(PAS)鉴定，并用细胞增殖核抗原(PCNA)免疫细胞化学检测了传代于鸡胚饲养层上PGC的增殖活性。培养的PGC经AKP和PAS鉴定均为阳性，AKP阳性为红棕色，PAS阳性为深红色，PCNA染色显示PGC集落为红棕色，表明PGC处于增殖状态。同时比较了不同体外培养条件(5%~20%胎牛血清(FCS), ITS培养液(M199+ 10 μg/mL 胰岛素+5 μg/mL转铁蛋白+3×10-8 mol/L亚硒酸钠), 条件培养液, 15%FCS+ITS, 15%FCS+40% 条件培养液)下PGC增殖情况。发现未经密度梯度离心的PGC生长较离心的PGC好，且5%FCS在原代培养中起较好的促增殖作用；而传代培养时在不同培养条件下，5%FCS组或ITS组形成的PGC集落的直径较大，而仅用单纯的条件培养液效果并不明显。结果表明除鸡胚饲养层可作为PGC增殖的支持体系外，在体细胞-PGC共培养方式下5%FCS或单独的ITS也可作为PGC体外增殖的一种培养体系。     

一株硫酸盐还原菌DSRBa的分离鉴定及特性分析

从内循环厌氧反应器颗粒污泥中分离、纯化得到一株硫酸盐还原菌命名为DSRBa,经形态和基于16SrDNA序列分析,该菌株归属于脱硫弧菌属（Desulfovibrio）。分别以甲酸钠、乙醇、乳酸钠、葡萄糖等为碳源,以硫酸盐、硫代硫酸盐、亚硫酸盐、单质硫为硫源,研究了不同温度、pH及不同硫酸根浓度对该菌株的影响。结果表明,菌株最适宜生长温度为30～35℃,最佳生长pH为7.0,无需绝对严格厌氧,当溶液中氧化还原电位（ORP）≤-40mV时,该菌株能较好生长,且生长4d后使溶液内氧化还原电位值达到-380mV,随后溶液内氧化还原电位基本保持不变。当系统内乳酸钠和酵母提取物浓度分别为3.5g·L-1和1g·L-1时,硫酸根浓度在1～4.5g·L-1范围对菌株生长无明显影响,且当SO24-浓度≤3g·L-1时,菌株生长4d对硫酸根的去除率达到90%以上。     

Isolation and Characterization of Atriplex hortensis and Sweet Chenopodium quinoa Starches

Starches from garden orach (Atriplex hortensis) and sweet quinoa (Chenopodium quinoa Willd. ‘Surumi’) seeds were isolated, examined for compositional characteristics, and compared with bitter quinoa (Cheno‐podium quinoa Willd.) starch. Garden orach and sweet quinoa seeds were similar in fat and ash contents, while garden orach seeds contained ≈10% more protein. Starches were isolated from seeds following a 12‐hr soak in dilute alkaline solution using a series of grinding, screening, centrifugation, and washing steps. Isolated starches viewed by scanning electron microscopy yielded angular, polygonal granules ≈1–2 μm in diameter. Starches displayed typical A‐type crystalline packing arrangements as determined by X‐ray powder diffractometry. Apparent amylose contents for garden orach (21.2%), sweet quinoa (20.6%), and bitter quinoa (19.8%) were determined according to colorimetric procedure. Differential scanning calorimetry data indicated a higher and wider gelatinization temperature range for garden orach as compared with sweet and bitter quinoa starches. Starch pasting profiles generated using a Rapid Visco Analyser indicated a reduced peak paste viscosity for garden orach starch relative to sweet and bitter quinoa and common corn starches.     

Isolation and Characterization of a Bensulfuron-Methyl-Degrading Strain L1 of Bacillus

The objectives of this study were to isolate a bensulfuron-methyl (BSM)-degrading strain of Bacillus spp. and to eval-uate its effectiveness in remediation of a BSM-contaminated soil. A BSM-degrading bacterium, strain L1, was successfully isolated in this study. Strain L1 was identified as Bacillus megaterium based on its morphological, physiological, and biochemical properties, G+C content, phylogenetic similarity of 16S rDNA, and fatty acid composition. Two experiments were used to examine BSM degradation by strain L1. When BSM was used as a sole carbon source in a mineral salt medium, the average degradation rate of BSM by strain L1 was 12.8%, which suggested that the strain was able to utilize BSM as a sole carbon and energy source. Supplement of yeast extract (200 mg L-1 ) significantly (P ≤ 0.01) accelerated the degradation of BSM by strain L1. Almost complete degradation (97.7%) of BSM could be achieved in 84 h with addition of yeast extract. In addition, in a sterile soil with 50 mg L-1 BSM, BSM degradation rate by strain L1 was 94.3% in 42 d, indicating the potential of using microbes for the remediation of BSM-contaminated soils in fields.