Role of major histocompatibility complex II antigen-presentation pathway genes in orange-spotted grouper infected with Cryptocaryon irritans

Cryptocaryon irritans, a pathogen model for fish mucosal immunity, causes skin mucosal and systematic humoral immune response. Where and how MHC II antigen presentation occurs in fish infected with C. irritans remain unknown. In this study, the full-length cDNA of the grouper cysteine protease CTSS was cloned. The expression distributions of six genes (CTSB, CTSL, CTSS, GILT, MHC IIA and MHC IIB) involved in MHC II antigen presentation pathway were tested. These genes were highly expressed in systematic immune tissues and skin and gill mucosal-associated immune tissues. All six genes were upregulated in skin at most time points. Five genes expected CTSS was upregulated in spleen at most time points. CTSB, CTSL and MHC IIA were upregulated in the gill and head kidney at some time points. These results indicate that the presentation of MHC II antigen intensively occurred in local infected skin and gill. Spleen, not head kidney, had the most extensive systematic antigen presentation. In skin, six genes most likely peaked at day 2, earlier than in spleen (5–7 days), marking an earlier skin antibody peak than any recorded in serum previously. This significant and earlier mucosal antigen presentation indicates that specific immune response occurs in local mucosal tissues.     

斜带石斑神经坏死病毒外壳蛋白基因克隆与序列分析

从患病毒性神经坏死病的斜带石斑鱼(Epinephelus coioids)的头部提取总RNA，根据已发表的神经坏死病毒外壳蛋白基因设计引物进行RT-PCR扩增，得到预期大小的基因片段。将此基因片段转入pET载体进行序列测定和分析，结果表明：编码斜带石斑神经坏死病毒(Orange-spoued grouper nervous necrosis virus，OGNNV)外壳蛋白基因的阅读框核苷酸数为1017bp，编码338个氨基酸；基因的核苷酸序列与野田村病毒科(Nodaviridae)的几种病毒的外壳蛋白基因序列比较结果显示，该病毒与p野田村病毒属(Betanodavirus)中的赤点石斑神经坏死病毒(red-spotted grouper nervous necrosis virus，RGNNV)的同源性最高(99％)，说明该病毒株是RGNNY血清型的成员。     

表面活性剂SDS与DBS对褐点石斑鱼急性毒性研究

[目的]研究表面活性剂SDS和DBS对水环境的污染程度,分析比较2类毒性数据处理方法的优劣。[方法]以褐点石斑鱼(Epinephelus fuscoguttatus)变态期仔鱼作为指示生物,采用静水法生物测试研究SDS和DBS的急性毒性。建立直线回归模型,并用非线性最小二乘拟合技术构建非线性回归模型,预测表面活性剂的毒性效应。[结果]2种表面活性剂的直线方程F检验均为极显著;其剂量—效应曲线(DRC)均可用双参数模型Weibull与Logit函数有效表征。直线和非线性回归模型对2种表面活性剂毒性效应估算表明,预测半致死浓度时,2种模型差异可忽略不计;预测极端效应浓度时,差异显著。直线回归模型估算的安全浓度SDS为0.4292mg/L,DBS为0.9543mg/L;双参数模型对48hLC50拟合预测值SDS为1.5033mg/L,DBS值为3.3416mg/L,两者预测结果一致,毒性均为SDS大于DBS。[结论]试验结果为研究表面活性剂污染对水环境造成的危害及评价提供参考资料,并为毒性数据的分析处理提供一种可供参考的新模式。     

Effect of dietary lipid level on growth performance,feed utilization and body composition of grouper <Emphasis Type="Italic">Epinephelus coioides</Emphasis> juveniles fed isonitrogenous diets in floating netcages

An experiment was conducted to determine the effect of dietary lipid levels on growth performance and body composition of grouper Epinephelus coioides juveniles cultured in floating netcages (1.5 m × 1 m × 1.5 m). Six isonitrogenous diets (53% dietary protein) with increasing dietary lipid concentration (5.16, 7.38, 9.73, 12.36, 14.13, 16.04% of dry material, DM) were fed to satiation to triplicate groups of 20 fish (mean weight: 10.9 ± 0.1 g) for 56 days. Dietary energy levels were 300, 322, 340, 371, 382, 400 kcal/100 g DM, respectively. Fish fed the 9L diet had the highest weight gain (WG) and specific growth rate (SGR), but they were not significantly different from that of fish fed the 7L or 12L diet (p > 0.05). FI varied inversely with dietary lipid levels. The poorest FCR and the lowest PER were observed in fish fed the 5L diet but these parameters showed no significant differences (p > 0.05) for fish fed the dietary lipid level ranging from 9 to 16%. Nitrogen intake decreased with dietary lipid levels. Fish fed the 7L diet showed the highest N gain, which was not markedly different from that of fish fed the 9L and 12L diets (p > 0.05). N retention was significantly lower in fish fed the 5L diet than in the other groups. Lipid intake and lipid gain increased with dietary lipid levels. Lipid retention (%) of the 7L diet was lowest but showed no significant differences among other treatments (p > 0.05). Proximate composition of fish body was significantly affected by dietary lipid level (p < 0.05). Body lipid content increased in direct proportion to dietary lipid levels. In contrast, moisture content of whole-body was correlated negatively to dietary lipid level. Dietary lipid content had no significant effect on protein content in whole-body and white muscle (p > 0.05). Condition factor (CF), hepatosomatic index (HSI) and viscerosomatic index (VSI) increased with increasing dietary lipid level. Based on second-order polynomial regression analysis of WG against dietary lipid level, a breakpoint of 10.0% was indicated to be the optimal dietary lipid concentration for maximum growth for grouper Epinephelus coioides juveniles cultured in floating netcages.     

配合饲料与蓝圆鲹饲养点带石斑鱼对水体中氮、磷含量的影响

分别以配合饲料与蓝圆鲹饲养点带石斑鱼（Epinephelus coioides）,观察其对养殖水体中氮、磷含量变化的影响。点带石斑鱼摄食后水体中的氨氮（NH3-N）、亚硝酸氮（NO2-N）、总磷（TP）含量随着其摄食后时间的延长呈现先升高后下降的趋势,NH3-N含量在鱼摄食后10-14h内达到高峰,NO2-N、TP含量在鱼摄食后14～18h达到高峰。试验组水体中的NH3N含量、NO2-N含量在排泄高峰期低于对照组（P〈0．01）,1TP含量在排泄高峰期也低于对照组（P〈0．05）。     

温度对云纹石斑鱼(Epinephelus moara)(♀)×七带石斑鱼(Epinephelus septemfasciatus)(♂)杂交F1胚胎发育和仔鱼活力的影响

用控温仪设定温度组,观察了在不同温度条件下(13℃、15℃、17℃、19℃、21℃、23℃、25℃、27℃、29℃、31℃)云纹石斑鱼(E.moara)(♀)×七带石斑鱼(E.septemfasciatus)(♂)杂交F1的胚胎发育情况,并对实验12 h后的好卵率、12 h后的胚胎发育时期、受精卵的孵化率和初孵仔鱼的畸形率进行了记录;仔鱼孵化出膜后,对其进行了耐饥饿实验,测定其每天的存活率和生存活力指数(Survival activity index,SAI)。实验结果表明,受精卵胚胎发育周期与温度呈正相关的关系,其受精卵孵化的适宜温度范围是17–25℃,温度低于17℃时,孵化率随温度的降低而降低,畸形率反之;温度为13℃时,不能孵化出仔鱼;温度高于25℃时,孵化率随温度的升高而降低,畸形率反之;仔鱼的SAI值随着温度的变化先升高后降低,在温度为17–21℃时,SAI值较高,分别为32.14±2.83、32.30±1.29、22.98±1.42,其他温度条件下SAI值均低于这3组。     

What causes cannibalization‐associated suffocation in cultured brown‐marbled grouper,Epinephelus fuscoguttatus (Forsskål, 1775)?

During culture of grouper, cannibalism is a frequent phenomenon that usually causes economic loss. Grouper culture often requires grading to prevent size‐dependent cannibalism. In comparison with orange‐spotted (Epinephelus coioides) and giant grouper (E. lanceolatus), failure to swallow prey during cannibalism is frequently observed in brown‐marbled grouper (E. fuscoguttatus). The cannibal cannot engulf the entire prey and the two fish ultimately end up dying together. Herein, we attempted to compare morphometric differences among orange‐spotted, giant and brown‐marbled grouper. We established a cannibal–prey total length (TL) relationship for brown‐marbled grouper: TL_prey=0.71 TL_cannibal−1.75. According to the equation, a difference of approximately 50% in TL is a threshold to use for grading the grouper. This value is much larger than that used (30%) in orange‐spotted and giant grouper, and this size difference results in a higher incidence of failed cannibalism attempts in brown‐marbled grouper. It is our belief that the standard grading method will fail to produce as good an effect in brown‐marbled grouper as is seen in orange‐spotted and giant grouper. Therefore, in addition to grading, other manipulations such as satiation feeding, nutritional supplementation and optimal stocking densities should be applied to reduce cannibalism of brown‐marbled grouper.     

鳙和棕点石斑鱼冷藏过程中ATP关联化合物的变化

为分析鳙和棕点石斑鱼贮藏过程中ATP关联产物含量的变化,本研究以高氯酸溶液作为蛋白沉淀剂,运用高效液相色谱法(HPLC),分析(4±1)°C和(0±1)°C贮藏条件下鳙和棕点石斑鱼体内6种ATP关联化合物含量的变化。结果显示,(4±1)°C条件下,2种鱼体内的ATP都快速降解;ADP、AMP含量则始终较低;IMP含量呈先快速上升后缓慢下降的趋势;鳙体内Hx R含量相对较高,棕点石斑鱼体内Hx R含量较少;Hx在4 d后即开始大量生成。在(0±1)°C条件下,2种鱼肉中ATP同样很快降解,ADP、AMP含量少;IMP降解速率变缓;2种鱼中Hx R生成和分解速率皆变缓;Hx生成速率较为平稳。鳙以IMP、Hx R为限速步骤,棕点石斑鱼以IMP为限速步骤。研究表明,不同的贮藏温度对ATP的降解影响较小,但对其关联产物的变化速率有较大影响,且不同鱼种间ATP关联产物降解规律也有所不同。     

点带石斑鱼溃烂病病原菌的分离与鉴定

溃烂病是我国北方海水工厂化养殖点带石斑鱼的主要病害之一。作者连续3a对天津地区的养殖点带石斑鱼溃烂病进行了流行病学调查,并从患病鱼病灶分离到7株致病菌,采用生理生化和分子生物学相结合的方法对7株致病菌进行了鉴定,结果表明,1株为杀鲑气单胞菌,5株为哈维氏弧菌,1株为河流弧菌。在连续3a的调查中,每年都分离到哈维氏弧菌,表明哈维氏弧菌是天津地区海水工厂化养殖点带石斑鱼溃烂病的主要条件致病菌,而杀鲑气单胞菌和河流弧菌是次要条件致病菌。研究结果将为控制石斑鱼溃烂病在海水工厂化养殖中的爆发和流行奠定基础。     

云纹石斑鱼和赤点石斑鱼杂交子一代线粒体相关基因的母性遗传特征分析

对云纹石斑鱼和赤点石斑鱼及其正反杂交子代的3种线粒体基因(COⅠ、16S rDNA、Cyt b)和核基因Tmo-4c4进行序列分析,其中16S rDNA序列中有明显的插入缺失位点,而其他3个基因序列无插入缺失变异。在12个分析样本中,COⅠ同源序列(387 bp)中共检测到41个核苷酸多态性位点,16S rDNA(529 bp)中有21个核苷酸多态位点,Cyt b(383bp)中有49个核苷酸多态位点。Tmo-4c4(467 bp)有8个核苷酸多态位点。序列差异分析和遗传距离比较结果显示,正反杂交子一代的3个线粒体基因序列与母本基因序列的同源性都为100%,与父本的基因序列同源性分别为COⅠ:90%和89.4%;16S rDNA:96%和96%;Cyt b:87%和87.2%。核基因Tmo-4c4正反交子一代与母本的序列同源性为98.9%~99.6%之间,与父本的同源性为98.7%~99.4%之间,没有明显的遗传差异。以上结果表明了云纹石斑鱼和赤点石斑鱼杂交子一代在3种线粒体基因上严格按照母性遗传的规律,而核基因Tmo-4c4没有明显的遗传偏向性。