不同卵色皱纹盘鲍(Haliotis discus hannai)幼鲍的生长与免疫比较

随机选取100只经过480 d养殖的3♀×10♂绿卵全同胞家系子一代(F1)的皱纹盘鲍(Haliotis discus hannai)幼鲍为实验材料,进行雌雄、卵色分类后,测量所有样本总重、壳长、壳宽等生长指标,并测定不同卵色幼鲍的超氧化物歧化酶(SOD)、碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、过氧化氢酶(CAT)和溶菌酶(LZM)的活性,进行单因素方差分析(One-way ANOVA).结果显示,在生长方面,不同卵色幼鲍的各生长指标无显著差异(P＞0.05),但绿卵幼鲍的各生长指标均高于其他3种卵色;在免疫酶活性方面,4种卵色幼鲍在SOD和CAT活性上无显著差异(P＞0.05),绿卵幼鲍的AKP、ACP和LZM 3种酶的活性与棕色和棕红色卵幼鲍存在显著差异(P＜0.05),而与棕绿色卵幼鲍无显著差异,其酶活性从高到低按卵色排列依次为绿色、棕绿色、棕色和棕红色.上述研究结果可为利用卵色选育皱纹盘鲍高产抗逆新品系提供依据.     

底栖硅藻种类组成对皱纹盘鲍匍匐幼体和稚鲍生长、存活的影响

为验证由生产培养获得的不同质量的底栖硅藻培养皱纹盘鲍不同生长阶段幼体的效果,采用经不同培养措施获得的两类底栖硅藻群落(饵料A和饵料B)培养皱纹盘鲍前期匍訇幼体和稚鲍.试验结果表明,饵料A培养前期匍匐幼体的存活率[(43.47±13.53)%]显著高于饵料B[(6.94±5.17)%],但生长指标差异不显著.两类饵料培养的稚鲍存活率差异不显著,但饵料B的生长速度和特定生长率[(0.151±0.025)mm/d和(4.706±0.492)%/d)]显著高于饵料A[(0.103±0.022)mm/d和(3.625±0.374)%/d)].说明皱纹盘鲍幼体在不同生长阶段适应底栖硅藻的种类不同.匍匐幼体期对底栖硅藻质量要求较高,稚鲍期由于食量增加和食性转化,对饵料数量的要求表现得更为明显.     

皱纹盘鲍一种丝氨酸蛋白酶抑制因子克隆与表达分析

通过RACE方法获得皱纹盘鲍丝氨酸蛋白酶抑制因子的cDNA序列.该序列全长1446个碱基,其中开放阅读框为410个碱基.该基因编码的氨基酸与丝氨酸蛋白酶抑制因子Serpin家族的结构相近.另外,在基因启动子区发现一些与免疫和癌症发生有关的转录因子结合位点.采用qRT-PCR方法分析该基因不同组织中表达情况,结果发现,在所有检测的组织包括外套膜、鳃、消化腺、血淋巴和肌肉中均有表达.另外,使用鳗弧菌感染并检测鳃组织中该基因表达情况,结果发现,细菌刺激后该基因表达量逐渐升高,表明丝氨酸蛋白酶抑制剂基因在皱纹盘鲍抗菌反应中能够发挥免疫调节作用.     

皱纹盘鲍消化道粘液细胞的类型与分布

采用阿新兰—过碘酸雪夫氏反应 (AB -PAS)染色方法显微观察和分析了皱纹盘鲍成贝消化道粘液细胞的类型与分布。据所显示颜色的不同 ,将粘液细胞分为Ⅰ—Ⅳ 4种类型 :分别呈红色、蓝色、紫红色和蓝紫色。口区唇上皮分布有大量粘液细胞 ,以Ⅱ型为主 ,另有少量Ⅳ型 ,主要为杯形并多为大型细胞。口球上皮及齿舌囊上皮粘液细胞分布密度高 ,以Ⅱ型为主 ,Ⅳ型次之。食道前段上皮有大量粘液细胞分布 ,中段和后段粘液细胞逐步减少 ,主要为Ⅱ型。肠各段上皮均有粘液细胞分布 ,但下行肠道比上行肠的粘液细胞相对多些 ,也以Ⅱ型为主 ,多为杯形和近圆形。直肠内、外上皮均有大量Ⅱ型粘液细胞分布 ,近圆形的较多。在嗉囊、胃盲囊和胃未见明显着色的粘液细胞     

皱纹盘鲍与盘鲍杂交技术的研究

采用皱纹盘鲍大连群体与盘鲍日本群体进行杂交育苗试验和鲍苗种生产。结果表明,皱纹盘鲍和盘鲍正、反交组合的受精率和孵化率均低于两亲本自交组,而其苗种成活率高于两亲本自交组（提高达20％以上）;皱纹盘鲍和盘鲍的正、反交组合鲍苗的生长情况是,其平均壳长明显高于两亲本自交组鲍苗的平均壳长,最长达2．01 cm ;皱纹盘鲍与盘鲍作为亲本进行鲍苗种生产,经160 d培育,共培育出鲍苗种2020．75万只（平均壳长2．01 cm ）,鲍出苗量平均4345．7只／m2。通过鲍种内杂交的育种方法,可以达到改良和优化鲍种质的目的。     

Effects of dietary guaiacol on shell biomineralization of juvenile abalone Haliotis discus hannai, Ino

A study was conducted to investigate the effects of guaiacol on shell biomineralization of abalone Haliotis discus hannai Ino. Seven semi-purified diets were formulated with graded levels of guaiacol (0, 5, 10, 50, 100, 500 and 1000 mg kg⁻¹). Abalone (initial weight: 0.29±0.01 g; initial shell length: 8.55±0.27 mm) were fed with these diets in a re-circulated water system for 152 days. The results showed that the contents of calcite in the shell increased significantly with dietary guaiacol levels, but the aragonite content decreased significantly. Dietary guaiacol significantly decreased the contents of calcium and strontium, but significantly increased the magnesium content in the shell. The effects of dietary guaiacol on the contents of phosphorus and zinc in shell were not significant. It was obvious that the ratio of potential acidic/basic amino acids [(Asx+Glx)/(Lys+Arg+His)] in the shell was elevated by dietary guaiacol. In conclusion, dietary guaiacol has effects on both the mineralogy and the amino acid composition in the shell. Furthermore, changes in mineralogy are related to the ratio of (Asx+Glx)/(Lys+Arg+His) in the shell.     

不同饵料对黑鲍幼鲍生长及存活的影响

本实验研究了裂片石莼(Ulva fasciata)、龙须菜(Gracilaria lemaneiformis)、海带(Laminaria japonica)、细基江蓠繁枝变种(Gracilaria tenuistipitata)、舌状蜈蚣藻(Grateloupia livida)和人工配合饲料6种饵料对黑鲍[(Haliotis discus hannai Ino)(♀)×(Haliotis discus discus Ino)(♂)]幼鲍存活和生长的影响.将6种饵料随机分成6组,每组3个重复,每个重复饲养黑鲍20只.实验幼鲍的初始体长、体质量分别为(12.29±0.03)mm、(0.223±0.002)g,实验为期35 d.实验结果如下:(1)不同饵料对幼鲍的存活率有显著影响(P＜0.05),且由大到小依次为海带组、人工配合饲料组、龙须菜组、蜈蚣藻组、江蓠组和石莼组.其中海带组、人工配合饲料组的存活率显著大于其他海藻组(P＜0.05);(2)不同饵料对幼鲍的生长有显著影响(P＜0.05),6种饵料组的幼鲍的特定生长率(SGR)由大到小依次为人工配合饲料组、海带组、江蓠组、石莼组、龙须菜组和蜈蚣藻组.其中人工配合饲料组、海带组的SGR显著大于其他饵料组(P＜0.05).壳长增长率、湿重增重率、干重增重率亦呈现相似趋势.实验结束时,蜈蚣藻组的体质量呈现负增长;(3)幼鲍对不同饵料的转化效率存在显著差异(P＜0.05),幼鲍对人工配合饲料的转化效率是最大.实验结果表明人工配合饲料和海带是黑鲍养殖生产上最佳的选择.     

温度胁迫对皱纹盘鲍生理和生化活动的影响

为探讨温度胁迫下皱纹盘鲍(Haliotis discus hannai Ino)的响应机制,采用室内控温实验,通过设置4个温度梯度(5℃、10℃、20℃和25℃),设计温度骤变处理组(皱纹盘鲍从15℃暂养温度直接转移至各实验温度)和温度缓变处理组(0.5℃/12 h),分析温度剧烈变化和温度缓慢变化对皱纹盘鲍耗氧率和排氨率的影响及其差异性;并对高温和低温处理下皱纹盘鲍消化腺中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、酸性磷酸酶(ACP)和溶菌酶(LSZ)活性的变化情况以及不同组织(血细胞和肌肉组织)中Cu/Zn–SOD基因的表达状况进行了研究。结果表明,皱纹盘鲍耗氧率和排氨率随海水温度的升高而增加,20℃达到最高值;25℃骤变处理组与缓变处理组耗氧率和排氨率间存在极显著差异(P0.01)。5℃和10℃骤变处理组皱纹盘鲍氧氮比(O/N)同缓变处理组间存在极显著差异(P0.01)。在高温胁迫后的3 h,消化腺中SOD、CAT和LSZ活性达到最高,而ACP活性在胁迫6 h后达到最高(P0.01);低温胁迫显著降低皱纹盘鲍LSZ的活性,于胁迫9 h后达到最低(P0.01)。不同温度胁迫下,皱纹盘鲍血细胞和肌肉组织中Cu/Zn–SOD基因的相对表达量均表现上调,与对照组存在显著差异性(P0.05)。本研究表明,温度胁迫能显著影响皱纹盘鲍的生理和生化活动,这将有助于探讨皱纹盘鲍夏季高死亡率的原因,为皱纹盘鲍健康养殖提供一定的理论依据。     

皱纹盘鲍与盘鲍杂交效果分析

从杂种优势的角度对皱纹盘鲍♀与盘鲍♂杂交子一代的经济性状进行分析。结果表明,4月龄杂交鲍子一代(F1)活体重比皱纹盘鲍增加36.87%,软体部干重增加33.21%,总干重增加29.20%,其它性状也显著优于皱纹盘鲍。杂交鲍F1的壳长、壳宽、活体重、软体部干重和总干重等主要性状的变异系数为12.94%~44.51%,其中软体部干重的变异系数最大,壳长的变异系数最小;且杂交鲍各性状的变异系数均大于皱纹盘鲍。杂交鲍子一代的变异有助于定向选择,提高杂交育种的效果。     

皱纹盘鲍卵受精时间对受精率及孵化率的影响

在22℃水温下，通过对皱纹盘卵子受精时间的实验，测定不同受精时间所对应的受精率及孵化率。结果表明，皱纹盘鲍卵子的受精时间不同，对其受精率，孵化率有显著影响，其最适的受精时间是2h以内。     

Effect of Feeding Regime on Compensatory Growth of Juvenile Abalone,Haliotis discus hannai,Fed on the Dry Sea Tangle,Laminaria japonica

Effect of feeding regime on compensatory growth of juvenile abalone (Haliotis discus hannai) fed on the dry sea tangle (Laminaria japonica) was determined. Thirty juvenile abalone averaging 15.7 g were randomly stocked into 18 50‐L plastic rectangular containers each. Six treatments were prepared in triplicate: Abalone were fed the dry sea tangle once a day at a satiation level with a little leftover for 16 wk as the control (Con) and other abalone were fed the dry sea tangle once a day at a satiation level with a little leftover for 15 wk after 1‐wk starvation (S1 treatment), 14 wk after 2‐wk starvation (S2 treatment), 13 wk after 3‐wk starvation (S3 treatment), 12 wk after 4‐wk starvation (S4 treatment), and 10 wk after 6‐wk starvation (S6 treatment), respectively. A linear relationship between weight change of abalone and wk of starvation was observed: Y (Weight of abalone) = ?0.17X (Wk of starvation) + 15.89 (R ² = 0.9462) (P < 0.0001). The highest survival of abalone was achieved in the S2 treatment, but not different from that of abalone in the Con, S1 and S3 treatments. Weight gain of abalone in the Con treatment was higher than that of abalone in the S4 and S6 treatments. Abalone fed on the dry sea tangle seemed to be able to achieve full compensatory growth up to 3‐wk starvation.     

磁化水对皱纹盘鲍生长影响的研究

水在磁场中可以被磁能活化,使水分子缔结链变小,活性增加。小分子水的渗透力、溶解力强,更容易穿过细胞膜,促进机体新陈代谢。本试验利用永磁铁使海水磁化激活,饲养皱纹盘鲍,试验结果显示,壳长2．0～2．5cm的幼鲍在三个月内增长率提高了1．3～1．6％,增重率提高了7％;饵料系数降低了1．7～2．1％;成活率提高了2．4～3．0％。     

碘伏对皱纹盘鲍的急性毒性试验

试验结果表明：碘伏对稚鲍2，42，48，72，96hLC50分别为3．91，0．64，0．38和0．36mg.L^-1有效碘。稚鲍对碘伏很敏感，且忍耐质量浓度范围较窄，不适合带水消毒。     

Effects of LED light quality on the growth,survival and metamorphosis of Haliotis discus hannai Ino larvae

Light is a key environmental factor influencing the growth, development and survival of aquatic organisms. We examined the effects of different light qualities (red, orange, white, blue, green or no light) and developmental stage at initial lighting [fertilized egg (FE), trochophore larva (TL), or eye‐spot larva (EL)] on the growth, development, and survival of larvae of the Pacific abalone Haliotis discus hannai Ino. Larva‐hatching success was significantly higher under blue, green, or no light compared with red, orange or white light (P < 0.05). Larval abnormalities were significantly increased under red, orange or white light compared with all other light qualities (P < 0.05). The incidence of metamorphosis in larvae illuminated from the TL stage was significantly higher under blue compared with other light qualities. Irrespective of the stage at initial illumination, the incidence of metamorphosis was lower in larvae cultured under red, orange or no light compared with other light qualities, but the differences were not significant (P > 0.05). Juvenile survival was significantly higher under blue or green compared with other light qualities (P < 0.05), with no significant effect of stage at initial illumination (P > 0.05). Larval size at completion of the shell was unaffected by stage at initial illumination, but was greater under blue or green light, while size at metamorphosis was greatest following illumination with blue or green light since the TL or EL stage (P < 0.05). Metamorphosis time was shortest with blue or green light and in cultures illuminated from the FE or TL stage (P < 0.05). Larval development from the FE to formation of the fourth tubule on the cephalic tentacles was fastest in larvae exposed since the FE or TL stage to blue or green light, compared with other light qualities (P < 0.05). However, there was no difference in terms of the rate of development from the FE to the TL stage between cultures lit or unlit since the FE egg stage (P > 0.05). These results suggest that a blue or green light source applied from the TL stage can increase the hatching and yield of H. discus hannai Ino, with important implications for the development of the aquaculture industry.     

Heat-shock protein70 gene expression in four hatchery Pacific Abalone Haliotis discus hannai Ino populations using for marker-assisted selection

Heat shock proteins (Hsps) are molecular chaperones that help organisms cope with stressful conditions. Here, we report on the growth rates and Hsp70 expressions in inbred and hybrid populations of abalone Haliotis discus hannai Ino. In abalone, inbred populations expressed more Hsp70 than hybrid populations at all temperatures, except at very high temperatures close to the physiological limit. At benign temperatures, there was a clear trend towards higher Hsp70 expression in inbred than hybrid populations, whereas at higher temperatures, a trend in the opposite direction was observed. The temperature of maximal Hsp70 expression (T_peak) varied with the population type. The T_peak of inbred populations (26°C) was lower than that of the hybrid populations (28°C). The maximal inducible Hsp70 of inbred populations was higher than that of hybrid populations. The results showed a trend towards higher expression in inbred population at a lower temperature. These results provide direct experimental evidence that hybrids can cope with the intrinsic stress even at non‐stressful temperatures. The constitutive Hsp70 may therefore be used for marker‐assisted selection in a breeding programme.     

皱纹盘鲍维生素D营养需要的研究

在以酪蛋白和明胶为蛋白源的精制饲料中添加不同浓度梯度的维生素D,并以海带为对照组饲喂皱纹盘鲍幼鲍103d。结果表明:幼鲍的生长及软体部水分,脂肪和蛋白质含量受到饲料中维生素D添加水平的影响显著,但成活率不受饲料中添加水平的影响;饲料中适量添加维生素D可以提高幼鲍软体部碱性磷酸酶的活力,但饲料中缺乏或过高水平的维生素D导致幼鲍软体部碱性磷酸酶活力的降低;贝壳中灰分及钙、磷含量受饲料中维生素D添加水平的影响显著。以增重,蛋白质增量和软体部碱性磷酸酶的活力为指标,幼鲍饲料中维生素D的适宜含量为100IU·(100g)-1饲料。     

4种多糖对皱纹盘鲍血细胞活性氧和血清凝集活力的影响

水温为(20.0±3)℃,盐度为30.0±1.5下,将灵芝多糖1、灵芝多糖2、脂多糖和酵母多糖分别配制成0、1.0、5.0、10、50、100、200μg·mL-1质量浓度,采用鲁米诺化学发光法(CL)和凝聚法检测其对皱纹盘鲍(HaliotisdiscushannaiIno)血细胞产生活性氧和血清凝集活力的影响。这些多糖均能增强鲍血细胞产生活性氧的能力和吞噬能力,其增强效果由强至弱依次为:灵芝多糖2>灵芝多糖1>脂多糖>酵母多糖;同种多糖中,急性组增强鲍血细胞活性氧产生和促进血清凝集活力的能力强于孵育组。本研究还比较了浸泡和注射灵芝多糖2(50μg·mL-1)和脂多糖(10μg·mL-1)对皱纹盘鲍血细胞活性氧产生和血清凝集活力的影响。结果表明,注射的效果优于浸泡。     

饵料对不同规格皱纹盘鲍能量收支的影响

对3种不同规格的皱纹盘鲍(41.40±2.05、54.22±2.66、63.17±2.52mm)分别进行不同饵料搭配的投喂,并对各生理活动的能量代谢进行了测量与计算。实验结果表明,搭配投喂的实验组中皱纹盘鲍能够摄取更多的有机物作为能量代谢的基质。扣除代谢能、排泄能和排粪能后,搭配投喂能提供给皱纹盘鲍的生长能显著高于对照组(P<0.05),尤其是孔石莼与裙带菜搭配投喂组以及裙带菜与海带搭配投喂组,其获得的生长能的比例在3种规格组中均处在很高的水平,是鲍的筏式养殖中值得推广使用的投喂方法。     

用流式细胞仪检测活体鲍倍性

分别取皱纹盘鲍（Ｈａｌｉｏｔｉｓ ｄｉｓｃｕｓ ｈａｎｎａｉ）足肌和血液制样,经ＤＡＰＩ染色后,用ＰａｒｔｅｃＰＡＳ－Ⅲ型流式细胞仪测定其倍性,试验证明三倍体鲍的ＤＮＡ相对含量是二保体的１．５倍,切足法和抽取血液法两种制样方法均得到满意结果,用流式细胞仪法检测鲍倍性适应鲍多倍体产业化的需要。     

Effects of light quality and intensity on the growth,survival and metamorphosis of Haliotis discus hannai Ino larvae

Light is a key environmental factor that influences the growth, culture and survival of aquatic organisms. This study examined the effects of different light qualities (red, orange, white, blue and green light) and intensities (5 μmol m^?2 s^?1, 15 μmol m^?2 s^?1 and 40 μmol m^?2 s^?1) on the hatching and metamorphosis of larva of Haliotis discus hannai Ino and the growth and survival of juveniles. It was found that under blue and green light, the hatching success rate and metamorphosis of larva were significantly higher than those for any other light quality (P < 0.05), and there was a decreasing trend with an increase in light intensity. Under red and orange light, the abnormality rate of trochophores in each light intensity treatment and the time required for the metamorphosis of larva were significantly higher than those for any other light group (P < 0.05). Both the abnormality and metamorphosis rates significantly increased with light intensity. Under white light, at an intensity of 40 μmol m^?2 s^?1, the settlement and size of larva at metamorphosis were significantly smaller than those for any other light intensity (P < 0.05). Under red and orange light, the SGR of juvenile abalone in each light intensity treatment was significantly lower than that in any other light group (P < 0.05), but no significant difference was identified (P > 0.05). Therefore, selecting blue and green light and controlling the light intensity to 5–15 μmol m^?2 s^?1 during fingerling reproduction of abalone should increase the hatching success rate of larva and the yield per water body.