黏红酵母表面黏附蛋白及肠黏液受体的初步研究

为获得对大菱鲆肠道具有较强黏附能力的益生菌株,了解其黏附机制,采用体外黏液黏附模型及Western-blot技术,对黏红酵母等4株益生菌的大菱鲆肠黏液黏附性能及黏红酵母表面粘附蛋白和肠黏液受体进行了初步研究.结果表明,4株益生菌均可黏附于大菱鲆肠黏液;但各菌株的黏附能力存在较大差异,其中黏红酵母黏附能力最强,鼠李糖乳杆菌次之,假丝酵母和枯草芽孢杆菌相对较低.黏红酵母表面蛋白预先与肠黏液孵育2h后,显著抑制了黏红酵母与肠黏液的结合,其黏附百分率下降了83.6%;但上述处理对其他3株益生菌的黏附没有明显影响.Western-blot显示黏红酵母表面蛋白中分子量为38.5ku和28.6ku的两个蛋白参与对肠黏液的黏附过程,大菱鲆肠黏液中分子量为27.3ku和22.3ku两个蛋白可与黏红酵母表面蛋白特异性结合.糖原染色显示上述4个蛋白均为糖蛋白.     

饲料中添加不同脂肪源、VC和VE对大菱鲆生长和非特异性免疫力的影响

研究了不同脂肪源、VC和VE以及上述3种复合营养性添加剂对大菱鲆非特异性免疫力、生长和存活率的影响。结果表明,以鳀鱼鱼油为脂肪源,添加高剂量的VE能提高大菱鲆血清补体活性,并且提高大菱鲆的溶菌酶活性,但不影响大菱鲆血液的中性粒细胞吞噬率及白细胞总数;饲料中VC含量增加到250mg/kg,大菱鲆血液中的白细胞总数明显增加,而进一步提高饲料中的VC含量时,其白细胞总数没有显著变化;血清溶菌酶活性在VC含量为250mg/kg时最高。饲料中VC含量对白细胞的吞噬活性没有影响。在75d的养殖时间内,摄食复合营养性免疫增强剂的大菱鲆,其体重增重率(平均为95·3%)和成活率(99·4%)均明显高于未摄食营养型免疫增强剂的对照组体重增重率(68·0%)和成活率(93·7%)。     

大菱鲆的摄食量与排空速率的初步研究

以常规实验生态方法,研究商品鱼养殖中大菱鲆的摄食量与排空率,得出个体体重约(420±65)g,在(18 6±0 5)℃日投喂1次的条件下,摄食量为15～30g/尾,平均摄饵量为(41 03±1 71)g/(kg·d),空胃率约11%,在同样条件下的排空周期约20h,初始排粪出现在投食后6h,排泄速率高峰出现于12～14h,排粪量为0 77～1 35g/(kg·h)。建议在适温期中,以鲜杂鱼为饵料的该鱼养成期的合理日投饵1次,投喂量以体重的4%为宜。     

大菱鲆消化酶的活力

以1龄大菱鲆(Scophalmus maximus)为研究材料，研究养殖水温下大菱鲆消化酶的分布及不同温度和不同pH条件下胃、幽门盲囊、前肠、中肠、后肠中蛋白酶、淀粉酶、脂肪酶的活性。确定饵料中主要营养成分的消化部位和主要消化酶的性质。结果表明，各部位蛋白酶活性的最适温度和pH依次为40℃，2．0；40℃，8．0；60℃，8.0；40℃，8.5；60℃，8．0。18℃养殖水温下，在适宜的pH范围内，各部位蛋白酶活力从高到低依次为幽门盲囊、胃、前肠、中肠、后肠；淀粉酶活性的最适温度和pH依次为40℃，6．5；40℃，8．0；40℃，7．5；50℃，7．0；40℃，8．0；淀粉酶活力从高到低依次为幽门盲囊、前肠、中肠、后肠、胃；脂肪酶活性的最适温度和pH依次为40℃，7．5；40℃，6．5；40℃，7.0；40℃，7.5；40℃，7.5；脂肪酶活力从高到低依次为前肠、中肠、幽门盲囊、后肠、胃。     

Surface disinfection of Atlantic halibut and turbot eggs with glutaraldehyde: evaluation of concentrations and contact times

To evaluate the effects of glutaraldehyde treatment at different disinfection temperatures, Atlantic halibut, Hippoglossus hippoglossus (L.), and turbot, Scophthalmus maximus (L.), eggs, incubated at 5 and 12°C, respectively, were disinfected with 400–1200 mg glutaraldehyde l-1 at three different contact times (2.5, 5 and 10 min). Egg batches of both poor and good quality were tested for halibut. Positive effects were more pronounced in poor than in good-quality batches at hatching. Egg disinfection had a highly positive effect on the viability of yolk-sac larvae in both types of batches. A level between 400 and 800 mg l-1 at a contact time of 5–10 min was optimal for halibut: at lower levels, the bactericidal effect was reduced, and at higher levels, there were indications of toxic effects. Halibut eggs disinfected with optimal doses of glutaraldehyde had furthermore a reduced hatching time and more synchronous hatching as compared with untreated eggs. Turbot was more sensitive to higher doses than halibut, and the best larval performance was obtained for 400–800 mg glutaraldehyde l-1 at a contact time of 2.5 min. A further evaluation should, however, be performed before recommendations are given for species incubated at temperatures higher than 5°C.     

Lipolytic activities in developing turbot larvae as influenced by diet

The development of neutral lipase and phospholipase activities was studied in larval turbot fed live prey. Activities of neutral lipase and phospholipase (activity larva⁻¹) increased significantly between days 6 and 24 after hatching in turbot larvae. The specific activities of both enzymes (activity μg protein⁻¹) decreased in older larvae. Feeding of a microdiet for 3 days (days 10–13) affected the lipolytic activity of neutral lipase and phospholipase negatively, compared to the larvae fed on rotifers. Since neutral lipase activities in whole larval homogenates and in the gut were significantly lower, it suggests a reduced synthesis rate and a reduced secretion of the enzyme in larvae fed the microdiet. A correlation between neutral lipase and phospholipase activities was found in larvae fed rotifers, but not in larvae fed the microdiet. This may indicate different regulating and stimulating mechanisms for these enzymes. The contribution of exogenous enzymes from ingested live prey to the total larval enzyme activity was about 6% for neutral lipase and 10% for phospholipase on day 6. The exogenous prey enzymes accounted for only 2% of the total activities in 12-day-old turbot larvae, suggesting that enzymes from prey did not contribute considerably to the digestion of lipids. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effect of an immunostimulating alginate on protein turnover of turbot (Scophthalmus maximus L.) larvae

High densities of marine larvae are often associated with microbial problems, mostly caused by opportunistic pathogens, resulting in poor growth and mass mortalities. The early life stages of fish have a very limited immune defence system. Therefore stimulation of non-specific immune mechanisms in fish larvae might be a very interesting tool. The present study evaluates the effects of a known immunostimulant on protein turnover in larval turbot (Scophthalmus maximus). Protein turnover and food intake was determined at day 13 using a time course after feeding the larvae with ¹⁵N-labelled rotifers. Turbot larvae fed with rotifers enriched with the immunostimulant FMI at first feeding had significantly higher fractional rates of protein synthesis when compared to a control group. This resulted in three-fold higher protein turnover in the larvae given the immunostimulant compared to the control group. Food intake, larval size and survival at the end of the experiment were similar in the two treatments. The effect of FMI in increasing protein turnover probably imply a higher larval viability and survival in case of environmental/disease stress. However, protein turnover is costly, and this may cause a trade-off between fast growth and stress-resistance/survival in fish larvae.     

Effect of some repetitive factors on turbot stress response

The stress response of the turbot, Scopthalmus maximus (L.), to repetitive factors including netting, air exposure, blood sampling and hand-stripping, were tested using two different tank sizes as well as two blood sampling techniques. Exposure of juvenile fish to air for 1-4 min had no immediate effect on plasma cortisol concentrations or haematocrit values. Similarly, the serial netting of immature fish from tanks did not significantly modify plasma cortisol concentrations, haematocrit or osmolarity values. Hand-stripping of mature males was more disturbing than air exposure. The cumulative effect on plasma cortisol levels and osmolarity of stress factors such as netting, air exposure, blood sampling and stripping applied simultaneously to mature males in a 16 m3 tank and repeated twice daily for several days was recorded. The level of cortisol increased from 5 ng ml-1 to 300 ng ml-1 after 10 days of treatment, while an osmoregulatory imbalance and fish death were observed. Moreover, adaptation of fish to smaller tanks seemed to improve the increased plasma cortisol levels and death rate. Reduction in the number of stress factors applied greatly decreased both the range of physiological responses and the death rate. In order to avoid a cumulative stress response, handling of fish should therefore be reduced to a minimum.