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东太湖常见沉水植物作为中华绒螯蟹饲料源的可行性研究

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本研究对东太湖常见的五种沉水植物(伊乐藻、苦草、马来眼子菜、金鱼藻 和轮叶黑藻)草粉的营养成分和重金属含量进行了分析,结果表明,沉水植物粗 蛋白为 17.99%~23.00%、粗脂肪为 3.37%~6.12%、灰分为 29.82%~34.71%、总糖为 8.49%~23.29%,氨基酸总量为 103.32~134.12 gkg -1 ,必需氨基酸为 39.43~49.60 gkg -1 。沉水植物还含有锰、铜、铁、锌、钾、钙等多种矿质元素,其中钙含量达 13.28~95.57 gkg -1 。沉水植物中铅、汞、砷、铬、镉等重金属含量均在我国渔用配 合饲料安全限量范围内。 根据伊乐藻、苦草、马来眼子菜、金鱼藻和轮叶黑藻等沉水植物营养成分组 成及相关研究成果,以 15%的添加量将上述沉水植物添加到基础饲料中,在室内 循环水池中饲养 9.37±0.15 g 的中华绒螯蟹(Eriocheir sinensis),共 8 周。分别测 定各组中华绒螯蟹的生长性能、免疫功能、肝胰腺组织结构和风味指标,为将沉 水植物作为中华绒螯蟹饲料源开发提供科学依据。 本研究的结果,①在生长性能上,中华绒螯蟹成活率由高到低顺序依次为, 伊乐藻组>轮叶黑藻组≈苦草组>金鱼藻组≈对照组>马来眼子菜组,组间差异不显 著(p>0.05);平均增重率从高到低顺序依次为,轮叶黑藻组>伊乐藻组>对照组> 苦草组>金鱼藻组>马来眼子菜组,马来眼子菜组中华绒螯蟹平均增重率显著低于 对照组(P<0.05),其余四种沉水植物组与对照组间差异不显著(p>0.05);饲料系 数从低到高顺序依次为,伊乐藻组<轮叶黑藻组<苦草组≈对照组<金鱼藻组<马来 眼子菜组,马来眼子菜组饲料系数显著高于对照组(P<0.05),其余四种沉水植物组 饲料系数与对照组差异不显著(p>0.05);各组肥满度和肝体比差异均不显著 (p>0.05)。②在中华绒螯蟹非特异性免疫功能与抗病力方面,各试验组的蟹血清 转氨酶活力与对照组无显著性差异(p>0.05);沉水植物组的蟹血清超氧化物歧化 酶活力显著高于对照组(p<0.05);轮叶黑藻组、伊乐藻组和苦草组蟹血清中的溶 菌酶活性显著高于对照组(p<0.05),金鱼藻组和马来眼子菜组也高于对照组,但 差异不显著(p>0.05);除马来眼子菜组降低外,其余沉水植物试验组中华绒螯蟹 对嗜水气单胞菌的抵抗力强于对照组。③从消化酶活性与组织结构上分析,除马 来眼子菜组外,沉水植物组的试验蟹都保持了较高的消化酶活性。肝胰腺组织学II 研究发现,各组蟹肝小管基膜完整,细胞结构正常,没有明显的病理变化;透射 电镜观察结果显示,对照组、伊乐藻组、苦草组和轮叶黑藻组蟹的肝细胞微绒毛 排列紧密且整齐,马来眼子菜组和金鱼藻组的肝细胞微绒毛较为稀疏,有部分已 经脱落,产生轻微的损伤。④饲料中添加沉水植物能增加中华绒螯蟹肌肉与肝胰 腺中游离氨基酸及呈味氨基酸的含量,其中,伊乐藻组与轮叶黑藻组达显著水平 (p<0.05)。沉水植物组试验蟹的肌肉与肝胰腺中 SFA 含量有所减少, MUFA、 PUFA 和 UFA 含量有所增加,但均未达到显著水平(p>0.05)。 本研究得出如下结论,①东太湖常见五种沉水植物草粉营养丰富,且达到渔 用饲料安全标准。②除马来眼子菜外,伊乐藻、苦草、金鱼藻和轮叶黑藻对中华 绒螯蟹的生长性能无明显影响,但可在一定程度下提高其免疫力和消化酶活力。 ③沉水植物加入饲料后主要是通过增加中华绒螯蟹肝胰腺和肌肉的游离氨基酸和 呈味氨基酸含量增强其风味。④东太湖轮叶黑藻、伊乐藻、苦草和金鱼藻等四种 常见沉水植物可作为中华绒螯蟹饲料源进行开发。 关键词,沉水植物;中华绒螯蟹;饲料源;东太湖 作 者,孙丽萍 指导老师,宋学宏III Assessment of common submerged macrophytes in East Taihu Lake as feed resources for Eriocheir sinensis Abstract The contents of nutritional components and heavy metals from five submerged macrophytes, Elodea canadensis, Vallisneria natans, Potamogeton malaianus, Ceratophyllum demersum L. and Hydrilla verticillata, in East Taihu Lake were determined. The results showed that the crude protein ranged from 17.99%~23.00%, crude fat from 3.37 to 6.12%, ash from 29.82 to 34.71%, total sugar from 8.49 to 23.29%, total amino acids from 103.32 to 134.12gkg -1 , and the essential amino acids from 39.43 to 49.60 gkg -1 in these five submerged macrophytes on the basis of dry weight. The results also revealed that a wide range of mineral elements, such as Mn, Cu, Fe, Zn, K, Ca were found in these plants, and Ca was as high as 13.28 ~ 95.57 gkg -1 . Even more importantly, lower contents of heavy metals, including Pb, Hg, As, Cr, Cd were detected in these plants, which met the national formula feed safety limit for aquatic animals. The dry powder of these plants were incorporated, respectively, into the basic diet at a level of 15% (w/w) to feed Chinese mitten crab (initial body weight: 9.37±0.15 g) every day for 8 weeks. The feeding experiment was conducted in an indoor recirculating aquaculture system. In order to provide scientific basis for submerged macrophytes as feed resources of Chinese mitten crab, body growth rate, immunity functions, hepatopancreas structure and flavor were analyzed. These feed macrophytes produced different effects on the survival rates of Chinese mitten crab, which were listed in the order of importance as E. canadensis > H. verticillata > V. natans > C. demersum L.> control group > P. malaianus, and there were no significant difference among these groups (p>0.05); The effects on the average growth rate were listed in the order of importance as H. verticillata > E. canadensis >IV control group > V. natans group > C. demersum L. > P. malaianus, and P. malaianus group was lower than the control significantly (p<0.05), the other submerged macrophytes groups were no significant difference compared with control (p>0.05). As for their effects on FCR, they were listed in decreasing order as E. canadensis > H. verticillata > V. natans > the control > C. demersum L. > P. malaianus and P. malaianus group was higherthan the control significantly (p<0.05), the other submerged macrophytes groups were no significant difference compared with control (p>0.05). With respect to the nonspecific immunity and disease resistance of Chinese mitten crab, No significant difference was found in transaminase activities in serum among all groups (p>0.05). The superoxide dismutase (SOD) activities of treatment groups with submerged macrophytes were higher than the control significantly (p<0.05). The activity of lysozyme (LSZ) from groups by feeding H. verticillata, E. canadensis, or V. natans, were higher than the control significantly (p<0.05), P. malaianus group and C. demersum L. group also present higher lysozyme activity than the control, but no significant difference was detected (p>0.05). In this condition, submerged macrophytes groups enhanced the resistance of Chinese mitten crab to Aeromonas hydrophila infection to a certain extent except P. malaianus group. The submerged macrophytes groups maintained a high digestive enzyme activity except P. malaianus group. The histology of hepatopancreas showed that basement membrane was complete, the structure of cell was normal, there were no obvious pathological changes in hepatopancreas. TEM results showed that the micro-villous structure of the control, E. canadensis group, H. verticillata group and V. natans group were tight and neat, but the microvilli of liver cells from P. malaianus group and C. demersum L. group were sparsely distributed, even some have been broken off, and caused slight damage. The incorporation of submerged macrophytes into basic diet could increased the levels of free amino acids and delicious amino acids in muscles and hepatopancreas of Chinese mitten crab, and supplements with E. canadensis and H. verticillata produced these effects significantly (p<0.05). No significant decrease in SFA or increase in MUFA, PUFA, UFA were found in muscles and hepatopancreas among all groups (p>0.05). It might be suggested that the main submerged macrophytes in East Taihu Lake areV rich in nutritional components, and all meet the safety standard for fish feed. Except P. malaianus, the other four submerged macrophytes had no effect on the growth of Chinese mitten crab, and they could enhance the immunity and digestion of Chinese mitten crab to a certain extent. H. verticillata, E. canadensis, V. natans could enhance the flavor of Chinese mitten crab by elevating the contents of free amino acids and delicious amino acids in muscles and hepatopancreas of Chinese mitten crab. All the tests showed that H. verticillata, E. canadensis, V. natans and C. demersum L. could be exploited as feed resources for Chinese mitten crab. Key words: submerged macrophytes; Eriocheir sinensis; feed resources; East Taihu Lake Written by: Ling-ping Sun Supervised by: Xue-hong Song目 录 前 言 1 文献综述 2 1 沉水植物的生态地位 2 2 沉水植物资源化利用研究进展 2 2.1 治理水体富营养化 3 2.2 用作肥料 3 2.3 用作燃料 3 2.4 药用或食用 4 2.5 用作饵料与饲料 4 2.6 其它方式 6 3 中华绒螯蟹的食性与营养需求的研究进展 6 3.1 中华绒螯蟹食性研究 6 3.2 中华绒螯蟹营养需求研究进展 6 4 研究目的和意义 8 试验一 东太湖常见沉水植物主要营养成分及安全性研究 9 1 材料与方法 9 2 结果 10 3 讨论 12 3.1 沉水植物中的营养成分及其含量 13 3.2 沉水植物作为饲料源开发的安全性 13 3.3 沉水植物作为饲料源开发的可能性 14 4 结论 14 试验二 东太湖常见沉水植物对中华绒螯蟹生长和免疫功能的影响 15 1 材料与方法 15 1.1 试验蟹 15 1.2 试验饲料 15 1.3 饲养管理 161.4 采样与分析方法 17 1.5 数据处理 19 2 试验结果 19 2.1 不同饲料对中华绒螯蟹生长性能的影响 19 2.2 不同饲料对中华绒螯蟹形体指标的影响 20 2.3 中华绒螯蟹常规营养成分的组成 21 2.4 不同饲料对中华绒螯蟹非特异性免疫力的影响 21 2.5 不同饲料对中华绒螯蟹的免疫保护作用 22 3 讨论 22 3.1 不同饲料对中华绒螯蟹生长性能的影响 22 3.2 不同饲料对中华绒螯蟹免疫功能的影响 23 4 结论 24 试验三 东太湖常见沉水植物对中华绒螯蟹消化酶活性及消化系统结构的影响 25 1 材料与方法 25 1.1 试验材料 25 1.2 采样与分析方法 25 2 试验结果 26 2.1 各试验组中华绒螯蟹肝胰腺消化酶活性的变化 26 2.2 各试验组中华绒螯蟹肠道消化酶活性的变化 27 2.3 不同饲料对中华绒螯蟹肝胰腺组织结构的影响 28 3 讨论 29 3.1 沉水植物对中华绒螯蟹肝胰腺和肠道消化酶活性的影响 29 3.2 沉水植物对中华绒螯蟹肝胰腺组织结构的影响 30 4 结论 30 试验四 东太湖常见沉水植物对中华绒螯蟹风味的影响 33 1 材料与方法 33 1.1 试验材料 33 1.2 试剂与仪器 33 1.3 试验方法 33 1.4 数据处理 342 试验结果 34 2.1 不同饲料组中华绒螯蟹肝胰腺和肌肉游离氨基酸组成和含量 34 2.2 不同饲料组中华绒螯蟹肝胰腺和肌肉中的脂肪酸组成和含量 36 3 讨论 40 3.1 不同饲料组中华绒螯蟹肝胰腺和肌肉中游离氨基酸的变化 40 3.2 不同饲料组中华绒螯蟹肝胰腺和肌肉中脂肪酸的变化 40 4 结论 41 全文结论 42