不同水平饲料镉对罗非鱼生长性能和肠道消化酶活性影响的研究

为研究不同水平饲料镉对罗非鱼生长性能和肠道消化酶活性的影响。将300尾罗非鱼随机分为5个处理,饲料镉添加水平分别为0、50、100、200、400 mg/kg,试验期为8周。结果表明：与对照组相比,饲料镉添加组增重率和特定生长率均逐渐降低,200、400 mg/kg添加水平组显著降低(P＜0.05);各处理组之间摄食率、饲料系数和成活率均无显著差异(P＞0.05)。饲料镉添加水平组罗非鱼肠道淀粉酶活性和蛋白酶活性均显著低于对照组(P＜0.05),当饲料中添加50 mg/kg镉时,罗非鱼肠道淀粉酶活性和蛋白酶活性即可显著降低(P＜0.05);罗非鱼脂肪酶活性仅在饲料镉添加水平400 mg/kg时显著低于对照组(P＜0.05),其余各处理组间差异不显著(P＞0.05)。表明罗非鱼对饲料镉耐受程度较高,饲料中镉达到200 mg/kg时,才可显著影响罗非鱼增重率和特定生长率。饲料中不同水平镉对罗非鱼肠道消化酶活性具有不同程度的抑制作用,饲料中添加50 mg/kg镉时即可显著降低肠道淀粉酶和蛋白酶的活性,肠道脂肪酶活性仅在400 mg/kg时显著降低。     

Impact of Sump Tank Size on Growth,Survival, and Production of Marron,Cherax tenuimanus,in an Intensive System

Juvenile marron, Cherax tenuimanus were cultured in an intensive crayfish culture system for a period of six months to study the effects of sump tank size on crayfish growth, survival, and production. Water volume in the large sump system was 2750 L and in the small sump tank system was 500 L. Crayfish were held in individual compartments (surface area 320 cm², depth 10 cm) and fed with commercial pellets (30% protein; 3-5% body weight per week). Specific growth rate (SGR) was 0.75%/day in the large sump system and 0.68%/day in the small sump system; survival was 83% in the large sump system and 71% in the small sump system; and biomass roduction (B) was 239 g/m² in the large sump system and 158 g/m² in the small sump system. These values were all higher (P < 0.05) in the large sump system. The results suggest that crayfish production could be improved by using large sump tanks in recirculating intensive crayfish culture systems.     

脱钙罗非鱼鱼鳞明胶提取工艺优化及其理化性质

文章以罗非鱼（Oreochromisnilotica）鱼鳞为原料,经乙二胺四乙酸二钠（EDTA—Na,）脱钙处理后采用酸法提取明胶。根据中心组合试验设计（centercombination of experimental design,CCD）原理展开试验,考察提取时间（h,X,）、提取温度（℃,X2）、浸酸时问（h,X1）和柠檬酸质量浓度（g·L-1,X）对明胶得率（％,y）的影响,并对所得明胶的理化性质与牛骨明胶进行比较。响应面分析结果表明,提取温度、提取时间和浸酸时问与明胶得率之间具有显著相关性;优化的提取条件为提取温度65℃、提取时间3．6h、柠檬酸质量浓度200g·L-1、浸酸时间11．3h。在此条件下的明胶得率为28．4％,与预测值30．1％相近。制得的鱼鳞明胶粗蛋白含量为89．9％,氨基酸组成中每1000总氨基酸残基中甘氨酸为352个,亚氨基酸（脯氨酸和羟脯氨酸）173个;凝胶强度高达245Bloom（g）,熔点为27．0℃,而比浓对数粘度及成胶质量浓度分别为0．030L·g-1。和8．0g·L-1。     

罗非鱼免疫球蛋白（IgM）重链基因全长cDNA序列分析

从已经构建的罗非鱼白细胞cDNA文库中测定获得罗非鱼免疫球蛋白（IgM）重链的eDNA序列,全长1885bp,有一个完整ORF阅读框长1770bp,5’UTR为29bp,3’-UTR为86bp,编码588个氨基酸。罗非鱼与大黄鱼、斜带石斑鱼、南极鳕鱼、乌鳢、鳜鱼、伯氏豚[鱼叚]虎鱼、牙鲆、花狼鱼、硬头鳟的IgM重链氨基酸序列有较高的同源性,最高达到69％,表明已经获得罗非鱼IgM重链基因;对所得氨基酸进行二级结构预测发现,罗非鱼IgM重链基因全长cDNA序列所编码的氨基酸分子量为41453．94Da。该试验结果为罗非鱼IgM重链基因功能的实验性鉴定工作奠定了基础。     

酸碱胁迫对奥尼罗非鱼幼鱼鳃组织结构及Na~+-K~+-ATP酶活力的影响

[目的]探讨酸碱胁迫对奥尼罗非鱼幼鱼鳃组织结构及Na~+-K~+-ATP酶活力的影响。[方法]将初始体质量(6.81±0.85)g奥尼罗非鱼(Oreochromis aureus×Oreochromis niloticus)幼鱼暴露于不同酸碱度(p H分别为4.5、6.0、7.5、9.0、10.5,依次设为试验Ⅰ、Ⅱ组、对照组及试验Ⅲ、Ⅳ组)的水体中,每组3个重复,每个重复30尾鱼。胁迫后0、12、24、48、72 h采集鱼鳃样本,采用HE染色石蜡组织切片及分光光度法比较酸碱胁迫对鱼鳃组织结构及其中Na~+-K~+-ATP酶活力的影响。[结果]对照组鱼鳃的组织结构未发现有明显病变的症状,且鳃中的Na~+-K~+-ATP酶活力维持在一定水平;各胁迫组鱼鳃组织结构均不同程度发生病变,出现鳃丝融合,动脉血管破损,鳃小片肿胀弯曲,顶端与末端粘充血和肿胀,黏液细胞增多,上皮细胞增生、脱落,泌氯细胞肿胀,细胞质空泡化等一系列病态症状,且其症状会随着酸碱程度的加深、暴露时间的延长而加重;酸碱胁迫后各胁迫组鱼鳃中Na~+-K~+-ATP酶活力均出现先升高后降低的变化,其中酸胁迫的峰值出现在48 h,而碱胁迫出现在12 h。[结论]该研究结果可为奥尼罗非鱼的养殖生产提供理论依据。     

罗非鱼无乳链球菌C5a肽酶(ScpB)的原核表达及其免疫原性

以尼罗罗非鱼(Oreochromis niloticus)为鱼体样本, 克隆了罗非鱼源无乳链球菌(Streptococcus agalactiae) scpB基因, 构建了原核表达质粒pET32a(+)-scpB, 转入大肠杆菌BL21(DE3)后, IPTG诱导表达, 表达的重组蛋白经镍离子金属螯合柱纯化及超滤管浓缩后, 进行SDS-PAGE和Western blot分析鉴定。纯化的重组蛋白以不同剂量(1 μg/g、3 μg/g、5 μg/g) 免疫尼罗罗非鱼后, 每周检测各实验组鱼体血清非特异性免疫指标[溶菌酶活性、超氧化物歧化酶(SOD)活力]及抗体水平变化情况, 并于免疫4周后以4倍LD₅₀的剂量对其进行人工攻毒。结果显示, 该重组蛋白在大肠杆菌中以包涵体的形式存在; 对尼罗罗非鱼的相对免疫保护率为69.66%~89.00%, 其中5 μg/g组的相对保护率最高; 受免鱼体血清中溶菌酶活性、SOD活力和抗体水平较对照组有极显著提高(P<0.01), 结果表明, 重组蛋白ScpB具有较强的免疫原性和保护作用。本研究旨在为GBS多肽疫苗的研制奠定基础。

     

Suppression effect of LHRH-A and hCG on Piwi expression in testis of Nile tilapia Oreochromis niloticus

Piwi proteins as constituent factors of the piRNA pathway are required for germline maintenance,meiosisand gonad development.Previous study showed hCG could regulate the Piwi expression in ovary of teleosts.In this study,we revealed effects of LHRH-A and hCG on Piwi expression in testis of tilapia usingReal-time PCR and Western blot.Both in vivo and in vitro study suggest that LHRH-A and hCG significantlydown-regulated Piwil-1 and Piwil-2 in mRNA or protein levels compared with controls.Meanwhile,tissueand cell distribution showed that Piwi proteins were mainly expressed in spermatocytes rather thanmature sperms.These results indicated thatHPG suppressesPiwiswhich mayplayacrucialrolein testisdifferentiation and development.     

First identification and characterization of Streptococcus iniae obtained from tilapia (Oreochromis aureus) farmed in Mexico

This is the first study to isolate, identify and characterize Streptococcus iniae as the causative disease agent in two tilapia (Oreochromis aureus) populations. The populations were geographically isolated, of distinct origins, and did not share water sources. Affected fish showed various external (e.g., exophthalmia and cachexia, among others) and internal (e.g., granulomatous septicaemia and interstitial nephritis, among others) signs. All internal organ samples produced pure cultures, two of which (one from each farm, termed S‐1 and S‐2) were subjected to biochemical, PCR and 16S rRNA sequencing (99.5% similarity) analyses, confirming S. iniae identification. The two isolates presented genetic homogeneity regardless of technique (i.e., RAPD, REP‐PCR and ERIC‐PCR analyses). Pathogenic potentials were assessed through intraperitoneal injection challenges in rainbow trout (Oncorhynchus mykiss) and zebrafish (Danio rerio). Rainbow trout mortalities were respectively 40% and 70% at 10⁴ and 10⁶ CFU per fish with the S‐1 isolate, while 100% mortality rates were recorded in zebrafish at 10² and 10⁴ CFU per fish with the S‐2 isolate. The obtained data clearly indicate a relationship between intensified aquaculture activities in Mexico and new disease appearances. Future studies should establish clinical significances for the tilapia industry.     

春夏季养殖罗非鱼体内食源性致病菌分析

[目的]为罗非鱼食源性致病菌的预警和防治提供理论基础。[方法]以2008年2～7月广东省不同区域罗非鱼养殖场为调查对象,定期采样分析罗非鱼体内及养殖环境中食源性致病菌的分布情况。[结果]春季罗非鱼养殖环境中共鉴定出8种致病菌,鱼体内共鉴定出6种致病菌;夏季罗非鱼养殖环境中共鉴定出13种致病菌,鱼体内共鉴定出12种致病菌;致病性嗜水气单胞菌、致泻大肠埃希菌、霍乱弧菌是养殖罗非鱼体内的主要食源性致病菌;夏季食源性致病菌的种类与数量多于春季。[结论]罗非鱼食源性致病菌的分布与养殖区域、水温及溶氧量等具有密切关系。     

海南粗榧内生真菌对罗非鱼无乳链球菌抑菌活性研究

利用活菌菌饼法和滤纸片法研究筛选64株海南粗榧内生真菌对罗非鱼源无乳链球菌的抑菌活性,并对活性菌株的代谢产物化学成分进行初步测试。菌饼试验结果显示,11株海南粗榧内生真菌对无乳链球菌具有明显抑制作用,占总菌株数的17.2％,抑菌圈在Φ(8.7±0.3～16.2±0.2)mm间。滤纸片试验结果显示,12株具有明显抑制作用,占总菌数的18.8％,抑菌圈在Φ（8.7±0.3～28.2±0.4）mm间。2种方法的最高活性菌株不一致,但最高活性菌株（B30、F52）和同时显示较高活性的菌株（B14、B30、B32、F7、F52）被测出都含有甾体及三萜类化合物。根据结果推测,对无乳链球菌有显著抑制作用的菌株可能含有甾体及三萜类物质。结果为深入研究内生真菌对罗非鱼无乳链球菌的抑制奠定基础,也为生物防治罗非鱼病害提供有益借鉴。