对虾抗菌肽转基因水稻抑制饲料腐败和防治罗非鱼细菌病害的初步研究

探讨了以植物生物反应器开发利用抗菌肽类物质防治水产动物病害的效果和机理。以携带中国明对虾抗菌肽—对虾素3-2的转基因水稻米糠制作罗非鱼饲料,研究其对饲料腐败和吉富罗非鱼嗜水气单胞菌肠炎的抑制效果。通过测定米糠和饲料中的霉菌总数、细菌总数,发现水稻中表达的对虾素3-2能有效抑制饲料中的霉菌与细菌的繁殖,对于保持饲料品质具有显著的效果。选取规格一致、体质健壮的吉富罗非鱼,随机分成5组,每组3个重复,分别投喂含20%非转基因米糠的饲料与含不同质量比(10%、20%、30%)的转基因米糠的饲料。通过嗜水气单胞菌攻毒保护实验,结果发现,转基因米糠对罗非鱼嗜水气单胞菌肠炎具有显著的保护效果。对吉富罗非鱼的肠道主要微生物数量、中肠石蜡切片的进一步分析发现,摄食转基因米糠饲料组的罗非鱼肠道内大肠杆菌比例降低,乳酸菌比例提高,肠道微绒毛的结构完整性显著改善。由此推断抗菌肽转基因水稻的防病效果可能与肠道微生物的改变和对肠道结构的保护作用有关。但是石蜡切片发现过量添加转基因米糠(30%)也会导致吉富罗非鱼肠道上皮细胞损伤。     

燃油植物橡胶树籽的开发利用

橡胶树是中国重要的经济树种,广泛种植于云南和海南等热带区域。橡胶籽是橡胶树的重要副产品,其种仁营养价值高;其中,粗蛋白含量约25%~30%,且必需氨基酸组成较平衡;粗脂肪含量为50%,且橡胶籽油成分近似于亚麻籽油,富含α-亚麻酸,其含量为花生油和葵花油的数十倍。橡胶籽中含有氢氰酸等抗营养因子,但经贮存、加热可钝化绝大部分氰化物。可见,橡胶籽及其加工产品可作为养殖动物饲料原料。前期研究发现：水产饲料中添加一定比例橡胶籽饼对鱼类生长和健康无明显负面影响,但高比例添加会对鱼类消化和健康产生一定负面影响,其可能与橡胶籽饼中高含量粗纤维有关;橡胶籽饼中赖氨酸和蛋氨酸含量相对较低,使用时应注意补充赖氨酸和蛋氨酸。橡胶籽油富含α-亚麻酸,是鱼油的优质替代源,可部分甚至完全替代水产饲料中的鱼油。本文综述了橡胶籽产品的营养价值及其在水产饲料中的应用现状,可为橡胶籽的开发利用提供理论基础。     

Biotransformation of tuna waste by co-fermentation into an aquafeed ingredient

Dried skipjack tuna ( Katsuwonus pelamis) waste (red meat, gills, viscera, fins, etc.) were mixed with 25% wheat flour and inoculated with a starter culture of Lactobacillus plantarum National Collection of Industrial Microorganisms (NCIM) 2912 (10⁸–10⁹ cells mL⁻¹) and Bacillus licheniformis MTCC 6824 (10⁷–10⁸ cells mL⁻¹). Changes in the nutritional quality (crude protein, crude fat, crude ash, crude fibre and nitrogen-free extract and aminoacids) were monitored during a fermentation period of 14 days. The proximate analysis showed significant changes in the composition of L. plantarum -fermented tuna (LPFT) and B. licheniformis -fermented tuna (BLFT) from the unfermented raw materials. Fermentation of tuna waste has resulted in a significant ( P <0.05) increase in the protein content of tuna waste between days 6 and 12. All the amino acid contents in BLFT increased during fermentation, whereas, in LPFT the levels of serine, histidine, tyrosine, methionine, cystine and phenylalanine contents were decreased. A marginal increase in calcium and phosphorus levels was recorded in the fermented products. The results of the study suggest that LPFT or BLFT can be used as a novel aquafeed ingredient for different fish species.     

Potential of indigenous Bacillus spp. as probiotic feed supplements in an extruded low‐fish‐meal diet for juvenile olive flounder,Paralichthys olivaceus

A 12‐week feeding trial was designed to assess the probiotic potential of indigenous Bacillus amyloliquefaciens and/or Bacillus subtilis singly or in combination with Bacillus licheniformis in an extruded feed for olive flounder (Paralichthys olivaceus) juveniles. A high fish meal (FM) diet (control) and a low‐FM diet containing an alternative protein blend (30% FM replacement, FM30) were formulated. Three other experimental diets were prepared by inclusion of B. amyloliquefaciens (BA), B. subtilis (BS), or a mixture of B. amyloliquefaciens, B. subtilis, and B. licheniformis (BASL) into FM30 diet, with a final concentration of 10⁶ CFU/g diet. Results indicated that the FM30 diet was well tolerated by flounder, and the overall performance was not affected by dietary treatments. Lysozyme activity and total immunoglobulin level were significantly reduced in flounders when fed with the FM30 diet compared with the BASL and BA diets, respectively. The Bacillus additives neither enriched the relative abundance of the corresponding Bacillus spp. in the relevant gut microbiota of olive flounder nor modulated the presumptive gene functions of the gut microbiome. Despite the absence of growth‐promoting effect, the tested probiotics could still be economically viable for use as immunostimulants in commercial flounder diets with partial FM replacement.     

Nutrient efficacy of microalgae as aquafeed additives for the adult black tiger prawn,Penaeus monodon

This study used a small amount of several lyophilized microalgae (5% of dry weight in aquafeed) to feed adult black tiger prawns Penaeus monodon for 4 weeks, aiming to promote prawn health and nutritional level. Apart from slight increases in ash content, chlorophyll and β‐carotene in the microalgae‐added feeds, there was no difference from the commercial diet in their biochemical composition. After 28 days of feeding, prawn body length, body weight, condition index and also total amount of fatty acids and amino acids of the flesh (2nd abdominal segment) were not affected by microalgae additives as compared with the control commercial diet‐fed group (P > 0.05). Prawns fed the microalgae diets, however, had a relatively better survival rate than those fed the commercial control diet, although no statistical significance was detected in any except for the diatom Melosira sp. The elevated survival rate was concomitant with a higher phagocytosis rate and superoxide dismutase (SOD) activity in the immune response analysis, and significantly a higher astaxanthin content detected in the tail muscle of microalgae diet‐fed prawns (5.36–10.78 ng of astaxanthin per gram of muscle DW). These findings demonstrated that low‐inclusion microalgal additives in prawn diets over a short feeding period (4 weeks) can result in healthier adult prawn, and hence could contribute to the development of an optimized feeding strategy for prawn aquaculture close to harvesting.     

Nutritional evaluation of two marine microalgae as feedstock for aquafeed

Using microalgae for animal nutrition provides an economically viable route for microalgae‐based technological innovation, especially in combination with CO₂ fixation given current global warming. However, this technology still lacks sufficient evaluation for screening microalgae for specific animals; meanwhile, current studies show some prejudice regarding ‘essential’ or ‘non‐essential’ ingredients. The results show that Dunaliella salina and Nannochloropsis salina were able to accumulate high protein (30%–57%) and lipid (20%–46%) content without affecting the performance of CO₂ fixation, which reached 0.28 and 0.23 g L^?1 day^?1 respectively. Both species exhibited high quality of lipids and proteins for Penaeus monodon based on the profiling. The essential fatty acid indexes (EFAI) for N. salina and D. salina were 3.81 and 9.02 respectively. Butyric acid was found to be present in both D. salina (12.03%) and N. salina (4.87%) based on the total fatty acids (FAs). The essential amino acid indexes (EAAI) for D. salina and N. salina were 2.23 and 1.29 respectively. Arginine was the most abundant essential amino acid (EAA) in both D. salina (10.83%) and N. salina (13.35%) on the basis of total amino acids (AAs). This study comprehensively compares the nutritional quality of the two commercial marine microalgae of D. salina and N. salina with the potential to be used as sustainable sources of lipids and proteins to reduce or even replace the traditional fish oil and fish meal in aquafeeds.     

Using Bacillus subtilis E20‐fermented soybean meal as replacement for fish meal in the diet of orange‐spotted grouper (Epinephelus coioides,Hamilton)

The potential of Bacillus subtilis E20‐fermented soybean meal (FSBM) as a partial alternative component of fish meal (FM) in fed diets of orange‐spotted grouper (Epinephelus coioides) was evaluated in this study. An FM‐based diet and seven diets containing 10%, 20% and 30% and 10%, 20%, 30% and 40% of FM replaced by soybean meal (SBM) and FSBM, respectively, were fed to grouper for 84 days to evaluate possible substitution levels of FM by tracking growth performance, digestive enzyme activity and morphological changes in the liver and distal intestine. No significant differences in survival and muscle composition of grouper were found between controls and treatments. Growth performance and feed efficiency of fish fed diets with FM replaced by FSBM up to 30% were not significantly different from controls, whereas significantly decreased growth performance and feed efficiency occurred with diets containing >20% of SBM. Based on the feed efficiency, the maximum substituted levels of FM by SBM and FSBM in grouper diets were 18.36% and 29.32%, respectively, based on broken‐line analyses. Histopathological changes in the liver and distal intestine, and significantly lower activity levels of digestive enzymes, including pepsin in the stomach and trypsin, chymotrypsin, amylase and lipase in the distal intestine, were found in fish fed a diet containing 30% of FM replaced by SBM. However, these parameters were improved by the substitution of FSBM. It is therefore believed that FSBM has great potential to be used as a protein source in grouper diets in partial replacement of FM.     

Recombinant neutral protease rNpI as fish feed additive to improve protein digestion and growth

The addition of heavy protein to aquafeed causes low digestibility, high cost and significant nitrogen pollution in the water. The addition of protease is an effective way of improving protein absorption, but few proteases have been developed specifically for aquafeeds. The hydrolysis efficiency of protease additives for plant protein in the neutral gut environment and their synergy with endogenous proteases has not been sufficiently investigated. In this study, the performance of the recombinant neutral protease rNpI as an aquafeed additive to reduce the raw protein materials in the diet of Carassius carassius during 56 days of growth was evaluated. rNpI effectively hydrolysed soya bean protein isolates (SPI) with a high degree of hydrolysation and a high small‐peptide yield. The rNpI‐treated SPI exhibited a notable antioxidant activity, which benefited fish health. In vitro digestibility indicated that rNpI complemented trypsin in SPI digestion better than papain or Alcalase. The growth trial showed that the addition of rNpI could warrant a reduction in the crude protein and gross energy of feed while producing a lower feed‐conversion rate (FCR) and better growth performance. Crucian that were fed an RM‐wheat‐E diet (5.5% rapeseed meal and 5% wheat reduction) showed the best performance, with the lowest FCR and highest weight gain. These results indicate that rNpI could be an effective additive for the aquafeed industry and could provide economic benefits by reducing aquafeed protein requirements and nitrogen emissions. This study provides novel information and strategies for additive protease use and development in aquafeed.     

液态和粉末脂肪对吉富罗非鱼幼鱼生长、健康及肌肉品质的影响

在目前的水产配合饲料生产中,油脂一般以液态形式添加到饲料原料中或者在真空环境中喷涂到饲料颗粒上,以脂肪粉进行应用的研究报道很少。本研究以豆油(SO,亚油酸∶亚麻酸=9∶1)和一种由豆油和菜籽油等组成的复合油(BO,亚油酸∶亚麻酸=5∶1)为脂肪源,分别以液态和粉末形式添加到饲料原料中制备4种配合饲料(SOl、BOl、SOp、BOp)。用其在室内水族箱中喂养初始平均体质量为15.38 g的吉富罗非鱼幼鱼8周后,各饲料投喂组鱼的增重率、特定生长率、饲料系数、肝体比、脏体比无显著性差异。与豆油饲料(SOl)投喂组鱼相比,SOp组的肝脏过氧化氢酶及超氧化物岐化酶活性显著升高、肌肉粗灰分含量显著降低,SOp、BOl、BOp组的血清甘油三酯和低密度脂蛋白水平、全鱼的水分含量都显著降低,肝脏总抗氧化能力、全鱼粗蛋白含量及肌肉剪切力都显著升高。此外,BOp组的血清过氧化氢酶活性及肌肉硬度显著高于SOl、BOl、SOp组。肝脏和肌肉的脂肪酸组成各组间差异不大,均反映饲料的脂肪酸组成。研究表明,与液态油相比,脂肪粉饲料对吉富罗非鱼幼鱼的生长无不良影响,且可改善鱼的健康及肌肉品质;复合油的效果优于豆油。结合脂肪粉具有包装、运输、储存、添加方便等优点,值得在水产养殖生产中进行推广应用。