赤点石斑鱼微卫星DNA的筛选

提取赤点石斑鱼(Epinephelus akaara)基因组DNA,经BamH I和Hind III双酶切并电泳回收300~1 000 bp的片段与经BamH I和Hind III双酶切的pUC18质粒重组,转化感受态大肠杆菌JM109,以α互补法筛选重组阳性克隆,建立赤点石斑鱼部分基因组文库。应用PCR混合池法筛选含核心序列为(CA)n和(GA)n微卫星位点的阳性克隆,经DNA测序,得到61个含微卫星序列的克隆,占总克隆数的2.89%。61个用于测序的克隆中共有111个不同类型的微卫星序列,其中(CA/TG)n重复类型57个,占51.4%;(AG/TC)n重复类型35个,占31.5%;其他重复类型19个,占17.1%。完美型、非完美型及复合型序列分别占48.7%、41.4%和9.9%。并在GenBank上注册了22个微卫星序列。     

斜带石斑鱼生长性能、体成分和组织游离氨基酸含量对饲料中牛磺酸含量的响应

本试验旨在探讨饲料中牛磺酸含量对斜带石斑鱼生长性能、体成分及组织游离氨基酸含量的影响。以酪蛋白和明胶为蛋白质源配制低牛磺酸含量的基础饲料,在基础饲料中分别添加0(对照)、0.4%、0.8%、1.2%、1.6%的牛磺酸,配制成5种试验饲料。将300尾平均体重为(19.14±0.06)g斜带石斑鱼幼鱼随机分为5组(每组3个重复,每个重复20尾),每组投喂1种试验饲料,饲养周期为56 d。结果表明:与对照组相比,各牛磺酸添加组试验鱼的增重率、摄食率和饲料效率均显著增加(P0.05),内脏指数显著降低(P0.05),肝脏指数、肥满度无显著变化(P0.05)。与对照组相比,各牛磺酸添加组试验鱼全鱼粗蛋白质含量显著升高(P0.05),全鱼粗脂肪含量显著降低(P0.05),全鱼水分和粗灰分含量则无显著变化(P0.05)。各牛磺酸添加组试验鱼肝脏和肌肉中总非必需氨基酸、总氨基酸、游离丝氨酸含量以及肝脏游离蛋氨酸含量均显著低于对照组(P0.05)。试验鱼血清、肝脏、肌肉和全鱼中牛磺酸含量随饲料中牛磺酸含量的增加而呈上升趋势,且与饲料中牛磺酸含量存在显著正相关关系(r0.907,P0.05)。随饲料中牛磺酸含量的增加,肝脏牛磺酸保留率呈上升趋势,而肌肉中牛磺酸保留率则呈先上升后下降的趋势;此外,各组试验鱼肌肉牛磺酸保留率均高于肝脏。由此得出,饲料中一定含量的牛磺酸能促进斜带石斑鱼生长和摄食,降低全鱼脂肪沉积,增加全鱼蛋白质沉积,降低组织中游离氨基酸含量。以增重率为指标,通过回归分析得出斜带石斑鱼饲料中牛磺酸的适宜含量为1.04%。     

凡纳滨对虾混养点带石斑鱼的研究

[目的]提高凡纳滨对虾的产量.[方法]利用生物防病原理,在凡纳滨对虾养殖中混养点带石斑鱼,研究其对凡纳滨对虾存活率和产量的影响.[结果]当凡纳滨对虾中混养点带石斑鱼密度为3000 尾/hm2左右时,凡纳滨对虾存活率和产量均达到最高,比单一养殖凡纳滨对虾存活率提高11.49％,平均产量提高3149.O kg/hm2.[结论]在凡纳滨对虾养殖中混养点带石斑鱼,能起到减少病害发生和降低养殖风险的作用.     

斜带石斑鱼甘露糖受体基因的克隆表达及其功能

为了研究斜带石斑鱼甘露糖受体(Epinephelus coioides mannose receptor,EcMR)在抗赤点石斑鱼神经坏死病毒(red-spotted grouper nervous necrosis virus,RGNNV)感染中的免疫功能,实验成功克隆与表达了EcMR.结果显示,EcMR cDNA全...     

哈氏弧菌诱导布氏石斑鱼头肾提取物的抗菌活性研究

以致病性哈氏弧菌（Vibrio harveyi）腹腔注射布氏石斑鱼（Epinephelus bleekeri）18h后,取布氏石斑鱼头肾制备头肾粗提物,并对该粗提物的抗菌活性进行研究,结果表明：布氏石斑鱼经哈氏弧菌诱导后,其头肾粗提物不仅对金黄色葡萄球菌（Staphyloccocus aureus）等革兰氏阳性菌有良好抑制作用,而且对大肠杆菌D31（Escherichio coli D31）、哈氏弧菌和溶藻弧菌（V.alginolyticus）等革兰氏阴性菌也有良好抑制效果;利用电泳凝胶琼脂糖弥散法检测发现,头肾粗提物中具有多种对大肠杆菌D31有较强作用效果的蛋白类抗菌物质;切胶回收AU—PAGE凝胶上的抗菌组分,利用SDS—PAGE凝胶检测各抗菌活性组分的分子质量大小依次约为：10．9KD,10．0KD,8．1KD,7．2KD,6．3KD,由于这些抗菌活性成分可被蛋白酶破坏,且分子质量均接近或低于10KD,可初步将其认定为抗菌肽。     

Possible application of mibolerone for induced sex inversion of grouper Epinephelus coioides

SUMMARY: Thirty immature juvenile grouper Epinephelus coioides (19–168 g bodyweight, BW) were randomly stocked in four units 6 t tanks to determine if mibolerone can be used to induce sex inversion in groupers. After acclimatization and weaning to artificial feed, the feed given daily (4% BW/day) was supplemented with 0, 50, 100, and 200 μg mibolerone/kg feed for about 18 weeks. Thereafter, the hormone treatment was withdrawn and the experiment was terminated at Week 24. Ten fish were killed for gonad histology at stocking to serve as an initial control while about three to five fish were killed every 8 weeks. In general, ovaries of initial controls showed the presence of moderate stromal cells and gonia and few primary oocytes. At Weeks 8 and 16, ovaries of the control fish (0 μg/kg) were similar to that of the initial control except that primary oocytes increased at Week 24. Gonads of fish fed diets containing 100 and 200 μg/kg had none to moderate spermatocytes and few spermatids at Week 8 and 16, although spermatozoa were not observed, indicating that the fish were undergoing spermatogenesis. Spermatogenesis at 50 μg/kg was not as advanced since only few spermatocytes occurred at Weeks 8 followed by moderate gonia and no spermatocytes and spermatids at Week 16. However, the presence of few primary oocytes was observed when mibolerone was withdrawn suggesting that sex-inversed fish reverted back to a female condition. These results show that sex inversion in juvenile grouper can be induced by oral administration of mibolerone and may have possible application on mature females to produce functional males.     

温度与盐度对云龙石斑鱼幼鱼耗氧率和排氨率的影响

为了研究饱食和饥饿状态下温度(24、27、30和33℃)、盐度(15、20、25、30和33)对云龙石斑鱼幼鱼(Epinephelus groupers)耗氧率(OCR)和排氨率(AER)的影响,采用密闭流水式呼吸实验方法对OCR和AER进行了测定。结果显示:饱食和饥饿状态下,温度、盐度对云龙石斑鱼幼鱼OCR和AER影响均显著(P0.05);当温度为24℃~33℃时,饱食状态下的OCR和AER分别比饥饿状态的提高30.29%~131.84%和131.82%~316.19%。饱食状态下,氧氮比(O∶N)、蛋白质供能比(PSR)、Q10(呼吸)、Q10(代泄)分别为12.54~20.18、27.74%~48.48%、1.21、1.60;而在饥饿状态下分别为25.77~35.90、19.49%~27.19%、1.87、3.41;当盐度为15~33时,饱食状态下OCR和AER分别比饥饿状态的提高100.41%~128.21%、182.19%~382.83%。饱食状态下,O∶N、PSR分别为13.32~20.72、39.47%~49.42%,而在饥饿状态下分别为29.18~30.98、22.59%~23.98%。研究表明,云龙石斑鱼幼鱼的最适生长温度为24℃~27℃,最适生长盐度为30~33。     

不同蛋白质源部分替代鱼粉对褐点石斑鱼幼鱼生长性能、体组成以及血清生化指标的影响

本试验旨在研究不同蛋白质源部分替代鱼粉对褐点石斑鱼幼鱼生长性能、体组成以及血清生化指标的影响.配制5组等氮等能饲料,对照组饲料(D1)含68％的鱼粉,试验组饲料(D2～D5)是在对照组饲料的基础上分别用豆粕替代10％的鱼粉、豆粕替代20％的鱼粉、啤酒酵母替代10％的鱼粉、玉米蛋白粉替代10％的鱼粉.选取体重在15 g左右的褐点石斑鱼幼鱼180尾,随机分为5组,每组3个重复,每个重复12尾.试验期为8周.结果表明:在生长性能方面,与对照组相比,D2、D3组的增重率、摄食率、特定生长率、粗蛋白质表观消化率差异不显著(P＞0.05),而D4、D5组的上述指标均显著降低(P＜0.05).与D2、D3组相比,D4组的特定生长率显著降低(P＜0.05),肝体指数显著升高(P＜0.05);D5组的摄食率、特定生长率、蛋白质效率和粗蛋白质表观消化率显著降低(P＜0.05),饲料系数、肝体指数显著升高(P＜0.05).在体组成方面,D5组全鱼和肌肉中粗蛋白质含量显著低于对照组和D2、D3组(P＜0.05);D4组肌肉中粗脂肪含量显著低于对照组和D2组(P＜0.05).在血清生化指标方面,D3组血清总蛋白、球蛋白、葡萄糖含量显著低于对照组(P＜0.05),而白球比显著高于对照组(P＜0.05);对照组、D4组血清甘油三酯含量显著高于D3、D5组(P＜0.05);各组间血清谷丙转氨酶和谷草转氨酶活性没有显著差异(P＞0.05).由此得出,作为鱼粉的替代品,豆粕比啤酒酵母和玉米蛋白粉能更好地满足褐点石斑鱼幼鱼的营养需求;从主要生长指标不受影响的角度考虑,褐点石斑鱼幼鱼饲料中可以用豆粕替代20％的鱼粉.     

复方中草药及其与抗生素联用对斜带石斑鱼溶藻弧菌病的治疗效果

通过药物体外抑菌效果及对斜带石斑鱼抗病力影响的研究,筛选有效治疗斜带石斑鱼溶藻弧菌病复方中草药与中西药联用配方。用诃子、白芍、甘草比例分别为0.9∶1.3∶0.9、1.2∶0.9∶1.0和1∶1∶1的复方中草药对溶藻弧菌进行体外抑菌实验;将质量分数为2.2%的复方中草药与中西药联用添加到基础饲料中设为实验组,研究中西药对斜带石斑鱼溶藻弧菌病治疗效果。复方中草药实验组Ⅰ、Ⅱ、Ⅲ为投喂饲料中添加不同比例诃子、白芍、甘草,中西药联用实验组Ⅳ、Ⅴ、Ⅵ分别为投喂饲料中添加不同比例诃子、白芍、甘草、恩诺沙星。结果显示,复方中草药配方二药物质量浓度200 mg/mL,为最佳配方。40 d病鱼死亡数量比较结果为对照组Ⅱ对照组Ⅳ对照组Ⅲ实验组Ⅲ实验组Ⅱ实验组Ⅰ实验组Ⅴ实验组Ⅵ实验组Ⅳ对照组Ⅰ。实验组Ⅰ投喂饲料加诃子、白芍、甘草比例为0.9∶1.3∶0.9的复方中草药,实验组Ⅳ配方为诃子、白芍、甘草、恩诺沙星比例为0.9∶1.3∶0.9∶1.3,分别为治疗斜带石斑鱼溶藻弧菌病效果最好的复方中草药和中西药联用配方。研究表明,中西药联用复方药效强于复方中草药,均能较好治疗斜带石斑鱼溶藻弧菌病。     

Histological observations of seasonal reproductive and lunar-related spawning cycles in the female honeycomb grouper Epinephelus merra in Okinawan waters

Seasonal reproductive and lunar-related spawning cycles of the female honeycomb grouper Epinephelus merra inhabiting Okinawan waters were examined by histological observation of ovaries. Gonadosomatic index (GSI) increased beginning in May and peaked in June. Histological observations revealed that many oocytes laden with yolk were present in the ovaries from May to August. From September to October, ovaries were occupied by immature oocytes. These results suggest that the reproductive season of E.   merra lasts for 4 months from May through August in Okinawan waters. When the fish were collected according to the lunar cycle, GSI increased with the approach of the full moon. Oocytes at various development stages were observed from the first quarter to the full moon. Fresh ovulatory follicles were present in the ovaries around the last quarter moon. These results suggest that E.   merra has a lunar spawning cycle and spawns between the full moon and the last quarter moon. Ovarian features of the fish collected around the last quarter moon were different among individuals; some fish had many oocytes at the tertiary yolk stage in the ovaries, while the ovaries of the others were occupied by the oocytes at the peri-nucleolus and the oil droplet stages. This observation suggests that a minor release of eggs occurs in this species before or after a major spawning lunar day. Oocytes at the migratory nucleus and the maturation stages were not observed in any ovarian samples. This may mean that maturation of oocytes is related to the spawning behavior of this species and makes rapid progress in the process of aggregation at the spawning sites.