鳗弧菌L-18株的限铁条件和铁调节外膜蛋白的免疫效果

利用Western-blot技术,对鳗弧菌L-18株的OMPs和IROMPs的免疫反应性进行了分析,并通过免疫牙鲆比较二者免疫保护力的差异,以期从IROMPs方向探索开发鳗弧菌新型鱼用疫苗,并为研究其免疫原理提供重要的科学依据。结果表明,培养基中加入不同浓度的联铁剂,鳗弧菌L-18株的生长和铁载体的表达出现规律性变化,其中加入100~130μmol.L-12,2′-联吡啶为最适合限铁条件,此时铁载体的表达量最高且生长受抑制程度较小。在此条件下,菌株表达出74.3 ku和77.4 ku的IROMPs,其中77.4 ku蛋白具有免疫反应性,为重要抗原。用全菌灭活疫苗、OMPs、IROMPs分别腹腔注射免疫牙鲆7周后攻毒,IROMPs疫苗的相对免疫保护力达到75.9%,远高于OMPs疫苗的51.8%。相对于OMPs,IROMPs的抗体效价显著提高,接近全菌疫苗组的水平,而血清杀菌活力没有显著变化。     

牙鲆的养殖与饲料

介绍了牙鲆亲鱼、苗种及养成各阶段的养殖方式、养殖条件发展情况，讨论了微颗粒饲料、硬颗粒饲料、膨化颗粒饲料、软湿颗粒饲料、鲜杂鱼生鲜料、粉末饲料等多种饲料在牙鲆的全养殖过程的应用情况，总结了牙鲆配合饲料及营养研究的进展情况，列出了行业标准中的牙鲆饲料的营养要求。     

牙鲆不同组织抗菌肽的提取及部分性质检测

以新鲜的牙鲆为试验材料,用100%丙酮和90%甲醇配制的提取液,从牙鲆的鳃、皮肤、肝、肾组织中提取抗菌肽,并对牙鲆抗菌肽粗提物进行部分性质检测。抑菌试验证明,牙鲆各组织提取物对大肠杆菌、鳗弧菌有较强的抑制作用,并且不同组织的抑制作用有所差别,而对金黄色葡萄球菌的抑制效果不明显。提取物经过加热处理后,抑菌圈略有变小。相反,经过胰蛋白酶处理后,抑菌圈略有变大。牙鲆的多种组织中含有对细菌有抑制作用的抗菌肽。     

饥饿对漠斑牙鲆前期仔鱼生长发育的影响

研究了饥饿对漠斑牙鲆前期仔鱼生长发育的影响,并计算了仔鱼的饥饿不可逆点。结果表明,仔鱼3日龄开口,卵黄于5日龄消耗完毕,油球于6日龄消耗完毕,混合营养期为3～6日龄,饥饿对仔鱼卵黄和油球的吸收速度影响不显著。摄食仔鱼生长呈线性增加,饥饿仔鱼则在饥饿开始一段时间后呈现负增长的趋势。10日龄饥饿仔鱼出现较为明显的饥饿特征,如胸角凸出,消化道萎缩等。在18～20℃培育水温下,仔鱼饥饿不可逆点出现在7日龄和8日龄之间。     

Stock structure of Japanese flounder inferred from morphological and genetic analyses

ABSTRACT:   Stock structure of Japanese flounder Paralichthys olivaceus has been inferred mainly from either morphological or genetic analyses. However, because the results of both analyses did not always agree with each other, an inclusive conclusion has never been obtained. In this study, the stock structure has been inferred from both morphological and genetic analyses using 722 wild Japanese flounder collected from nine locations along the Japanese coast. The dorsal and anal fin ray counts were larger in the southern than in the northern populations. In total, 1041 bp of mitochondrial NADH dehydrogenase subunit-2 ( ND2 ) and 1830 bp of ND5 sequences were aligned. There are 578 variable sites in the concatenated sequence from the two genes, which defined a total of 490 haplotypes. Both results of morphological and genetic analyses indicated that the western Kyushu group, which included the Nagasaki and Kagoshima populations, was divided from the other seven populations. This is the first report to reveal the heterogeneity of the western Kyushu group based on statistical analysis.     

Effects of temperature changes on the endogenous rhythm of oxygen consumption in the Japanese flounder <Emphasis Type="Italic">paralichthys olivaceus</Emphasis>

ABSTRACT:   The effects of temperature changes on the endogenous rhythm of oxygen consumption in the Japanese flounder Paralichthys olivaceus which were acclimated to 3, 14 or 20°C in advance were investigated. The oxygen consumption rate (OCR) of individuals (total length >28 cm) that were acclimated to 3°C, maintained a relatively low value of 6.9 ± 2.3 mL (mean ± SD) O₂/kg ww (wet weight)/h till 5.8°C, which is considered a 'minimum' rate. Thereafter, the OCR gradually increased up to 18.7 mL O₂/kg ww/h at the temperature of 10.6°C. The OCR of individuals acclimated to 14°C stabilized with 124.2 ± 24.1 mL O₂/kg ww/h, when tested at the relatively constant temperature of 14.4 ± 0.17°C. Maximum entropy spectral analysis revealed that the peak OCR occurred at 23.9 h intervals, which could be a circadian rhythm. The mean OCR showed a peak at 06.00–08.00 hours. For the temperature above 25°C in which the fish were acclimated to 20°C, the rhythmic patterns of OCR were observed until the experimental temperature reached 26.4°C. For the temperature above 28.4°C, the metabolic activity rhythm of the OCR was dampened and then the OCR abruptly rose again. It is assumed that the oscillator of the endogenous rhythm lost the governing of normal metabolic activity in Japanese flounders above 28.4°C. These results suggest that the temperatures of around 5.8, 26.4 and 28.4°C are the 'lower limit temperature', 'upper incipient lethal temperature', and the 'critical thermal maximum', respectively, for the endogenous rhythm of oxygen consumption in the Japanese flounder.     

牙鲆致病性迟钝爱德华菌毒力基因的检测及序列分析

为探讨牙鲆致病性迟钝爱德华菌毒力基因携带与分布,以毒力基因为分子靶标研究其致病机理及建立快速检测方法,根据 GenBank 上的基因序列,设计2对引物扩增致病性迟钝爱德华菌毒力基因 fimA和小肠结肠炎耶尔森菌毒力基因 irp2,结果在7个供试牙鲆病例的130株致病性迟钝爱德华菌中,irp2毒力基因的阳性率为46．15％（60／130）,fimA 毒力基因的阳性率为100％,且均与已发表的参考菌株相应序列高度同源,同源性分别为97．32％和97．59％。可见耶尔森菌强毒力岛（HPI）在牙鲆致病性迟钝爱德华菌中广泛分布,但不同病例分离菌株间存在差异;fimA 毒力基因存在于所有的被检致病性迟钝爱德华菌中,可以作为快速检测致病性迟钝爱德华菌的标志物。     

Effect of feeding ratio on growth and body composition of juvenile olive flounder Paralichthys olivaceus fed extruded pellets during the summer season

The effect of feeding ratio on growth and body composition of juvenile olive flounder fed extruded pellets was determined during the summer season. Thirty juvenile olive flounder (initial body weight of 17 g) per tank were distributed into 21, 180-l flow-through tanks. Seven treatments that included triplicate groups of feeding ratio in 5% decrement were prepared for this study: 100% (satiation), 95%, 90%, 85%, 80%, 75% and 70% of satiation. Fish in the control group were hand-fed to apparent satiation twice a day. Then feed allowance in the rest of the six groups was determined based on average feed consumption in the control group. The feeding trial lasted for 7 weeks. Survival was over 97% and was not significantly (P > 0.05) affected by the feeding ratios. Weight gain and SGR of fish fed to 100% of satiation were not significantly different from those fed to 95% of satiation but significantly (P < 0.05) higher than those of fish fed to 90%, 85%, 80%, 75% and 70% of satiation. Weight gain, SGR and feed consumption of flounder linearly (P < 0.001) decreased with a decrease in feeding ratio. However, feed efficiency ratio, protein efficiency ratio and protein retention in fish body were not significantly (P > 0.05) affected by the feeding ratio. The crude protein levels of the whole body without liver or liver were significantly affected by the feeding ratio. Hepatosomatic index and condition factor of fish were not significantly (P > 0.05) affected by the feeding ratio. It can be concluded that optimum feeding ratio for growth of juvenile olive flounder could be lowered to 95% of satiation without growth suppression.     

Optimum dietary protein level for maximum growth of juvenile olive flounder Paralichthys olivaceus (Temminck et Schlegel)

This experiment was conducted to determine the optimum dietary protein level for juvenile olive flounder Paralichthys olivaceus (Temminck et Schlegel) fed a white fish meal and casein‐based diets for 8 weeks. Olive flounder with an initial body weight of 4.1 ± 0.02 g (mean ± SD) were fed one of the six isocaloric diets containing 35%, 45%, 50%, 55% and 65% crude protein (CP) at a feeding rate of 4–5% of wet body weight on a dry‐matter basis to triplicate groups of 20 fish per aquarium. After 8 weeks of feeding, per cent weight gain (WG) and feed efficiency ratios of fish fed the 55% CP diet were not significantly higher than those from fish fed the 50% and 65% CP diets, but significantly higher than those from fish fed the 35% and 45% CP diets. Fish fed the 50%, 55% and 65% CP diets had significant higher specific growth rates than did fish fed the 35% and 45% CP diets; however, there was no significant difference among fish fed the 50%, 55% and 65% CP diets. The protein efficiency ratio was inversely related to the dietary protein level; that is, maximum efficiency occurred at the lowest dietary protein level. Broken‐line model analysis indicated that the optimum dietary protein level was 51.2 ± 1.8% for maximum weight gain in juvenile olive flounder. The second‐order polynomial regression analysis showed that the maximum WG occurred at 57.7% and it revealed that the minimum range of protein requirement was between 44.2% and 46.4%. These findings suggest that the optimum dietary protein level for maximum growth could be greater than 46.4%, but less than 51.2% CP in fish meal and casein‐based diets containing 17.0 kJ g^?1 energy for juvenile olive flounder.     

容器颜色对漠斑牙鲆白化率和成活率的影响

本文就培养容器的颜色对漠斑牙鲆稚鱼的白化率和生长率的影响进行初步研究。结果表明:用深蓝色和黑色等深色容器培养的稚鱼白化率显著低于白色和黄色等浅色容器(p<0.05),黑色、深蓝色、黄色和白色4个组别(N=100)的白化率分别为20.8%、14.4%、57.6%和86.6%;白色最高为86.6%,深蓝色最低为14.4%;而用不同颜色的容器培养漠斑牙鲆的稚幼鱼,对其成活率影响不大(p>0.05),黑色、深蓝色、黄色和白色4个组别的成活率分别为96.0%、90.0%、82.5%、93.0%;白化稚幼鱼的生长发育并无异常。