【作者】 周文；

【导师】 向建国；

【作者基本信息】 湖南农业大学 ， 农业硕士（专业学位）， 2022， 硕士

【摘要】 棘胸蛙（Quasipaa spinosa）药食同源,被誉“百蛙之王”,但因其蝌蚪变态期成活率低限制了该产业的发展。棘胸蛙蝌蚪变态期成活率主要受营养、疾病、发育、环境应激等因子的影响。营养不良可能导致棘胸蛙蝌蚪发育不良、畸形率高、肝病频发甚至死亡。目前尚无棘胸蛙蝌蚪专用饲料,一般用其它饲料替代,造成变态高死亡率和畸形率。本文分别对棘胸蛙蝌蚪3个时期,即Ⅰ期（后肢芽发育26～30期）、Ⅱ期（五趾分化31～40期）、Ⅲ期（变态41～46期）进行了营养成分测定,了解不同时期棘胸蛙蝌蚪的营养需求水平,设计了11个不同蛋白或脂肪水平的饲料(植物蛋白V_35%、V_40%、V_45%、V_50%;动物蛋白A_35%、A_40%、A_45%、A_50%;脂肪F_14%、F_19%、F_23%),用2种黑斑蛙蝌蚪商品饲料（CK₁、CK₂）做对比,探讨了不同营养水平对棘胸蛙蝌蚪生长发育及肝功能的影响,以期为开发棘胸蛙蝌蚪专用饲料,实现精准投喂和健康养殖提供理论参考。主要结论有:（1）Ⅰ、Ⅱ、Ⅲ期棘胸蛙蝌蚪鲜重蛋白质含量分别为8.23%、9.47%、10.6%;脂肪含量分别为6.00%、3.30%、6.10%;碳水化合物含量分别为0.90%、3.10%、1.20%;∑EAA/∑AA为46.77%,其中∑BACC占∑EAA的41.85%;∑PUFA占∑FA的0.4222%,其中C_18:2n6c占∑PUFA的89.48%。（2）35%～50%范围内的植物蛋白水平高低对变态成活率无显著影响（P>0.05）,均在97.65%以上;高植物蛋白有利于棘胸蛙蝌蚪生长发育,V_45%、V_50%时棘胸蛙蝌蚪的生长指数均优于其它各组;随植物蛋白水平的增加,SOD、LZM活性在45%达到峰值,而AST、ALT、CAT、T-AOC活性显著下降（P<0.05）,MDA活性显著升高（P<0.05）,过高的植物蛋白水平会导致棘胸蛙蝌蚪肝脏的抗氧化能力、代谢能力和防御体系受损,不利于肝脏的健康。棘胸蛙蝌蚪适宜的植物蛋白水平为40%左右。（3）35%～50%范围内的动物蛋白水平高低对棘胸蛙蝌蚪的生长无显著影响（P>0.05）,但影响变态成活率,A_50%组幼蛙变态成活率仅21.54%,显著低于其它各组（P<0.05）;随动物蛋白水平增加,棘胸蛙蝌蚪肝脏SOD、LZM活性在40%达到峰值,AKP活性45%达到峰值（P<0.05）,CAT、T-AOC、AST活性显著下降（P<0.05）,MDA、ALT活性显著升高（P<0.05）,高动物蛋白影响肝脏的抗氧化能力、代谢能力和防御体系,肝脏的健康状况受到影响。棘胸蛙蝌蚪适宜的动物蛋白水平为35%左右。（4）动、植物蛋白水平均为35%时,变态成活率无显著差异（P>0.05）;相同蛋白水平时,植物蛋白组幼蛙的体重、体长以及肝脏酶活性显著高于动物蛋白组（P<0.05）,但畸形率显著高于动物蛋白组（P<0.05）。综合分析棘胸蛙幼蛙的变态成活率、畸形率、肝脏健康状况发现,过高的蛋白水平会影响棘胸蛙蝌蚪肝脏的抗氧化力、代谢能力和防御体系,适宜蛋白水平35%～40%;棘胸蛙蝌蚪期不宜使用单一的蛋白源,纯动物蛋白源会降低变态成活率,纯植物蛋白源会增加幼蛙畸形率,早期使用植物蛋白为主的动植物蛋白源,后期使用动物蛋白为主的动植物蛋白源。（5）在45%动物蛋白水平时,不同脂肪水平对棘胸蛙幼蛙变态成活率、畸形率、肝体比、脏体比、胴体比、MDA活性均无显著影响（P>0.05）,但F_14%组的SOD、LZM、AKP、CAT、T-AOC活性显著低于其它各组,而AST、ALT活性显著高于其它各组,F_23%组的LZM活性显著高于其它组（P<0.05）,适宜范围内随脂肪水平提高棘胸蛙蝌蚪肝脏的抗氧化和防御能力增强,但肝脏的代谢能力会受到影响。棘胸蛙蝌蚪适宜的脂肪水平在19%左右为宜。更多还原

【Abstract】 Quasipaa spinosa has the same origin as medicine and food,and is known as the"King of Frogs",but the low survival rate of tadpoles during metamorphosis limits the development of the industry.The survival rate of Q.spinosa tadpoles during metamorphosis was mainly affected by factors such as nutrition,disease,development and environmental stress.Malnutrition may lead to stunted growth,high deformity rate,frequent liver disease and even death in Q.spinosa tadpoles.At present,there is no special feed for the frog tadpoles,and it is generally replaced by other feeds,resulting in high mortality and deformity rate of metamorphosis.In this paper,the three stages of the tadpoles of Q.spinosa were measured,namely stageⅠ（stage 26-30 of hindlimb bud development）,stage Ⅱ（stage 31-40 of five-toed differentiation）,and stage Ⅲ（stage of metamorphosis 41-46）,to understand the nutritional requirements of tadpoles in different periods,and designed 11 feeds with different protein or fat levels（Plant protein V35%,V40%,V45%,V50%;Animal protein A35%,A40%,A45%,A50%;Fat F14%,F19%,F23%）,compared with 2 kinds of commercial feeds（CK₁,CK₂）of Q.spinosa tadpoles,to explore the effects of different nutritional levels on the growth and development and liver function of Q.spinosa tadpoles.It provides a theoretical reference for the development of special feed for the frog tadpoles and the realization of precise feeding and healthy breeding.The main conclusions are:（1）The fresh weight protein contents of the tadpoles of stageⅠ,Ⅱand Ⅲ were 8.23%,9.47%and 10.6%;the fat contents were 6.00%,3.30%and 6.10%;the carbohydrate contents were 0.90%,3.10%and 1.20%,respectively.%;∑EAA/∑AA is46.77%,of which∑BACC accounts for 41.85%of∑EAA;∑PUFA accounts for 0.4222%of∑FA,of which C_18:2n6caccounts for 89.48%of∑PUFA.（2）The level of plant protein in the range of 35%～50%had no significant effect on the survival rate of metamorphosis（P>0.05）,all above 97.65%;high plant protein was beneficial to the growth and development of tadpoles,V_45%,V_50%The growth index of Q.spinosa tadpoles at was better than other groups;with the increase of plant protein level,the activities of SOD and LZM reached the peak at 45%,while the activities of AST,ALT,CAT and T-AOC decreased significantly（P<0.05）.the activity of MDA was significantly increased（P<0.05）.Excessive levels of plant protein would damage the antioxidant capacity,metabolic capacity and defense system of the liver of Q.spinosa tadpoles,which is not conducive to the health of the liver.The suitable plant protein level of the frog tadpole is about 40%.（3）The level of animal protein in the range of 35%～50%had no significant effect on the growth of tadpoles of the frog chestnut（P>0.05）,but affected the survival rate of metamorphosis.The survival rate of metamorphosis in the A_50%group was only21.54%,a significant lower than other groups（P<0.05）;with the increase of animal protein levels,the activities of SOD and LZM in the liver of Q.spinosa tadpoles reached a peak at 40%,and the activity of AKP reached a peak at 45%（P<0.05）.The activity of AST was significantly decreased（P<0.05）,and the activities of MDA and ALT were significantly increased（P<0.05）.High animal protein affected the antioxidant capacity,metabolic capacity and defense system of the liver,and the health of the liver was affected.The suitable animal protein level of the frog tadpole is about 35%.（4）When the levels of animal protein and vegetable protein were both 35%,there was no significant difference in the survival rate of metamorphosis（P>0.05）;at the same protein level,the body weight,body length and liver enzyme activity of frogs in the vegetable protein group were significantly higher than those in the animal protein group.group（P<0.05）,but the deformity rate was significantly higher than that of the animal protein group（P<0.05）.Comprehensive analysis of the metamorphosis survival rate,deformity rate,and liver health status of Q.spinosa young frogs found that excessive protein levels would affect the antioxidant capacity,metabolic capacity and defense system of the liver of Q.spinosa tadpoles,and the optimal protein level was 35%～40%.It is not advisable to use a single protein source in the tadpole stage of Q.spinosa.Pure animal protein source will reduce the survival rate of metamorphosis,and pure plant protein source will increase the deformity rate of juvenile frogs.In the early stage,plant protein-based animal and plant protein sources are used,and animal protein sources are used in the later stage.Protein-based animal and plant protein sources.（5）At 45%animal protein level,different fat levels had no significant effect on metamorphosis survival rate,deformity rate,liver-to-body ratio,visceral-body ratio,carcass ratio,and MDA activity in juvenile Q.spinosa（P>0.05）.However,the activities of SOD,LZM,AKP,CAT and T-AOC in the F14%group were significantly lower than those in the other groups,while the activities of AST and ALT were significantly higher than those in the other groups,and the LZM activity in the F23%group was significantly higher than that in the other groups（P<0.05）,with the increase of fat level within the appropriate range,the antioxidant and defense capacity of the liver of Q.spinosa tadpoles enhanced,but the metabolic capacity of the liver would be affected.The appropriate fat level of the frog tadpole is about 19%.更多还原