黄粉虫酶饲料、虫粪培养基及其β-蜕皮激素的研究
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摘要
黄粉虫(Tenebrio molitor L.)属鞘翅目拟步甲科粉甲属。既是大型仓储害虫,又是重要的科研实验材料,也是极具有开发潜力的资源昆虫。本试验主要研究了纤维素酶、半纤维素酶和木聚糖酶3种酶饲料对黄粉虫幼虫生长发育的影响,黄粉虫虫粪培养基对食用菌生长发育的影响及黄粉虫p-蜕皮激素提取的方法和条件。获得以下主要研究结果：
1.酶饲料饲养黄粉虫的效果
(1)利用麦麸与稻糠、水花生、玉米秆、三叶草、水稻秆按1：1混合饲喂30～40日龄黄粉虫幼虫得出：稻糠与麦麸按照1：1质量混合是作为全麦麸替代饲料的最佳选择,其饲喂30天后平均鲜重为0.0246克/头、平均重增为0.0188克/头、干物质含量为39.98%,食物消化率、转化率和利用率分别为63.55%、28.20%和17.93%。
(2)利用稻糠与麦麸分别以1：4、2：3、1：1、3：2、4：1的比例混合饲喂40～50日龄黄粉虫幼虫得出：稻糠与麦麸按2：3质量比例的混合饲料是作为全麦麸饲料替代品的最佳选择,其饲喂30天后平均鲜重为0.04915克/头、平均重增为0.03329克/头、食物消化率、转化率和利用率分别为62.34%、29.74%和19.50%。
(3)利用pH试纸和pH计两种方法测定黄粉虫幼虫中肠液的pH值,测得结果分别为4.3694和4.412,得出黄粉虫幼虫中肠环境的pH值约为4.4。
(4)利用分别加入纤维素酶、半纤维素酶和木聚糖酶的酶饲料饲喂50-60日龄黄粉虫幼虫得出：①酶含量为0.8%纤维素酶饲料对黄粉虫幼虫生长有明显的促进作用,其饲喂30天后平均鲜重为0.07015克/头、平均增重为0.05549克/头,食物消化率、转化率和利用率分别为64.04%、31.41%和26.72%；②酶含量为1.0%半纤维素酶饲料对黄粉虫幼虫生长有明显的促进作用,其饲喂30天后平均鲜重为0.06724克/头、平均增重为0.05251克/头,食物消化率、转化率和利用率分别为61.60%、44.78%和26.69%；③酶含量为0.8%木聚糖酶饲料对黄粉虫幼虫生长有明显的促进作用,其饲喂30天后平均鲜重为0.06595克/头、平均增重为0.05133克/头,食物消化率、转化率和利用率分别为63.24%、37.88%和23.95%。
2.黄粉虫虫粪培养基对食用菌生长的影响
(1)试验得出黄粉虫虫粪浸提液的最佳的制备方法是：虫粪与水25：100混合,70℃下浸提16h。
(2)利用黄粉虫虫粪浸提液固体和液体培养基培养平菇、金针菇、灵芝及香菇结果表明：平菇、金针菇、灵芝、香菇分别在虫粪浸提液含量为25%、20%、25%、20%的平板培养基中菌丝生长最佳。在以黄粉虫虫粪浸提液为氮源的液体培养基中,金针菇、灵芝、香菇在虫粪浸提液含量为20%时,球菌生长最佳；平菇则在其含量为25%时球菌生长最佳。
(3)利用黄粉虫粪培养生产食用菌的结果表明：黄粉虫粪能够培养香菇、平菇、金针菇、茶树菇、鸡腿菇、灵芝这6种食用菌；培养基中虫粪含量越高,金针菇菌丝生长越快；添加虫粪的培养基可使金针菇和平菇增产。
3.黄粉虫p-蜕皮激素提取的方法
利用Box-Behnken响应面分析法分析不同温度与时间互作对黄粉虫幼虫β-蜕皮激素提取率的影响,并建立回归模型表。得出最佳浸提条件为：溶剂乙醇、温度80℃、时间3h,β-蜕皮激素提取率为0.0563%；此外试验还得出最佳的浸提料液比为虫重/乙醇容积=30：100,提取率为0.0848%。
Tenebrio molitor L. belong to Tenebrio of Tenebrionidae in Coleoptera. They are not only large-scale storage pests and important research materials, but also the extremely potential resources. This test mainly studied on the influence of Tenebrio molitor's growth from Cellulose, Hemicellulase and Xylanase, the influence of edible fungi's growth from Tenebrio molitor's feces medium as well as the methods and conditions of P-molting hormone extraction in Tenebrio molitor.
1.Preliminary results from Tenebrio molitor's feeding of different Enzyme fodders
(1) After feeding 30～40-day-old Tenebrio molitor by mixing the wheat bran and rice chaff, Alternanthera philoxeroides, corn stem, Trifolium L., rice stem, wheat bran in the ratio of 1:1, the result was obtained:the rice chaff could be the best choice on replacing wheat bran as feedstuff. After feeding for 30 days, the average fresh weighted 0.0246 g/larvae, the average fresh increasing by 0.0188 g/larvae, dry matter content for 39.98%, AD for 63.55%, ECD for 28.20%, and ECI for 17.93%.
(2) After feeding 40～50-day-old Tenebrio molitor by mixing the wheat bran and rice chaff in the quality ratio of 1:4,2:3,1:1,3:2 and 4:1, the result was obtained:the best choice is mixing the wheat bran and rice chaff in the ratio of 2:3 as feedstuff instead of the whole wheat fodder. After feeding for 30 days, the average fresh weighted 0.04915g/larvae, the average fresh quality increasing by 0.03329g/larvae, AD for 62.34%, ECD for 29.74%, and ECI for 19.50%.
(3) Using the two methods by pH test strips and pH-meter to determinate the pH value of Tenebrio molitor's intestines, the results were 4.3694 and 4.412 respectively. So the conclution is that the pH value of intestinal environment in Tenebrio molitor is about 4.4.
(4) Using the Enzyme fodders, which were joined with Cellulose, Hemicellulase and Xylanase respectively, to feed 50～60-day-old Tenebrio molitor, the conclution were as follows:①The 0.8% cellulose fodders could promote Tenebrio molitor's growth obviously After feeding for 30 days, the average fresh weighted 0.07015g/larvae, the average fresh quality increasing by 0.05549g/larvae, AD for 64.04%, ECD for 31:41%, and ECI for 26.72%.②The 1.0% Hemicellulase fodders could promote Tenebrio molitor's growth obviously. After feeding for 30 days, the average fresh weighted 0.06724g/larvae, the average fresh quality increasing by 0.05251g/larvae, AD for 61.60%, ECD for 44.78%, and ECI for 26.69%.③The 0.8% Xylanase fodders could promote Tenebrio molitor's growth obviously. After feeding for 30 days, the average fresh weighted 0.06595g/larvae, the average fresh quality increasing by 0.05133g/larvae, AD for 63.24%, ECD for 37.88%, and ECI for 23.95%.
2. The influence of edible fungi's growth from Tenebrio molitor's feces mediums
(1) The optimal preparation method of Tenebrio molitor's feces leaching solution through the test is as follows:leaching 16h on the condition of 70 degrees after mixing the feces and water by the ratio of 25:100.
(2) Cultivating Pleurotus ostreatus(Fr.)Kummer, FlammuLina velatiper(Fr.) Sing, Ganoderma lucidum (W.curt.:Fr.) Karst and Lentinus edodes (Berk.)Sing by use of solid and liquid mediums from Tenebrio molitof s feces leaching solution, the results show that:the hypha growth of the four mentioned mushrooms was the best in the flat mediums with Tenebrio molitor's feces leaching solution respectively being 25%,20%,25%,20%. In the liquid medium that used Tenebrio molitor's feces leaching solution as nitrogen source, the hypha growth of FlammuLina velatiper(Fr.) Sing, Ganoderma lucidum (W.curt.:Fr.) Karst and Lentinus edodes (Berk.)Sing was optimal with Tenebrio molitor's feces leaching solution being 20%; While the hypha growth of Pleurotus ostreatus(Fr.)Kummer was optimal with 25%.
(3) The result of using Tenebrio molitor's feces to cultivate and product edible fungis shows that:the Tenebrio molitor's feces could cultivate Lentinus edodes (Berk.)Sing, Pleurotus ostreatus(Fr.)Kummer, Flammulina velatiper(Fr.) Sing, Agrocybecha Sing Hnang, Copyinds comatus(MUⅡ.Fr) Gray and Ganoderma lucidum(W.curt.:Fr.) Karst; The content of feces is higher, the Flammulina velatiper (Fr.) Sing hypha growth faster; The production of Flammulina velatiper (Fr.) Sing and Pleurotus ostreatus(Fr.)Kummer could increase with the Tenebrio molitor's feces.
3.Preliminary studies on the methods of P-molting hormone extraction in Tenebrio molitor
This test used analysis method of Box-Behnken to analysis the effect on the extraction rate of P-molting hormone in Tenebrio molitor from interactions between time and temperature, and establish regression model table. The optimal Leaching condition was obtained:leaching 3 hours with ethanol on the condition of 80 degrees, with the extraction rate being 0.0563%. Besides, this test also obtained the optimal leaching ratio between material and liquid is 30:100, with the extraction rate being 0.0848%.
1.酶饲料饲养黄粉虫的效果
(1)利用麦麸与稻糠、水花生、玉米秆、三叶草、水稻秆按1：1混合饲喂30～40日龄黄粉虫幼虫得出：稻糠与麦麸按照1：1质量混合是作为全麦麸替代饲料的最佳选择,其饲喂30天后平均鲜重为0.0246克/头、平均重增为0.0188克/头、干物质含量为39.98%,食物消化率、转化率和利用率分别为63.55%、28.20%和17.93%。
(2)利用稻糠与麦麸分别以1：4、2：3、1：1、3：2、4：1的比例混合饲喂40～50日龄黄粉虫幼虫得出：稻糠与麦麸按2：3质量比例的混合饲料是作为全麦麸饲料替代品的最佳选择,其饲喂30天后平均鲜重为0.04915克/头、平均重增为0.03329克/头、食物消化率、转化率和利用率分别为62.34%、29.74%和19.50%。
(3)利用pH试纸和pH计两种方法测定黄粉虫幼虫中肠液的pH值,测得结果分别为4.3694和4.412,得出黄粉虫幼虫中肠环境的pH值约为4.4。
(4)利用分别加入纤维素酶、半纤维素酶和木聚糖酶的酶饲料饲喂50-60日龄黄粉虫幼虫得出：①酶含量为0.8%纤维素酶饲料对黄粉虫幼虫生长有明显的促进作用,其饲喂30天后平均鲜重为0.07015克/头、平均增重为0.05549克/头,食物消化率、转化率和利用率分别为64.04%、31.41%和26.72%；②酶含量为1.0%半纤维素酶饲料对黄粉虫幼虫生长有明显的促进作用,其饲喂30天后平均鲜重为0.06724克/头、平均增重为0.05251克/头,食物消化率、转化率和利用率分别为61.60%、44.78%和26.69%；③酶含量为0.8%木聚糖酶饲料对黄粉虫幼虫生长有明显的促进作用,其饲喂30天后平均鲜重为0.06595克/头、平均增重为0.05133克/头,食物消化率、转化率和利用率分别为63.24%、37.88%和23.95%。
2.黄粉虫虫粪培养基对食用菌生长的影响
(1)试验得出黄粉虫虫粪浸提液的最佳的制备方法是：虫粪与水25：100混合,70℃下浸提16h。
(2)利用黄粉虫虫粪浸提液固体和液体培养基培养平菇、金针菇、灵芝及香菇结果表明：平菇、金针菇、灵芝、香菇分别在虫粪浸提液含量为25%、20%、25%、20%的平板培养基中菌丝生长最佳。在以黄粉虫虫粪浸提液为氮源的液体培养基中,金针菇、灵芝、香菇在虫粪浸提液含量为20%时,球菌生长最佳；平菇则在其含量为25%时球菌生长最佳。
(3)利用黄粉虫粪培养生产食用菌的结果表明：黄粉虫粪能够培养香菇、平菇、金针菇、茶树菇、鸡腿菇、灵芝这6种食用菌；培养基中虫粪含量越高,金针菇菌丝生长越快；添加虫粪的培养基可使金针菇和平菇增产。
3.黄粉虫p-蜕皮激素提取的方法
利用Box-Behnken响应面分析法分析不同温度与时间互作对黄粉虫幼虫β-蜕皮激素提取率的影响,并建立回归模型表。得出最佳浸提条件为：溶剂乙醇、温度80℃、时间3h,β-蜕皮激素提取率为0.0563%；此外试验还得出最佳的浸提料液比为虫重/乙醇容积=30：100,提取率为0.0848%。
Tenebrio molitor L. belong to Tenebrio of Tenebrionidae in Coleoptera. They are not only large-scale storage pests and important research materials, but also the extremely potential resources. This test mainly studied on the influence of Tenebrio molitor's growth from Cellulose, Hemicellulase and Xylanase, the influence of edible fungi's growth from Tenebrio molitor's feces medium as well as the methods and conditions of P-molting hormone extraction in Tenebrio molitor.
1.Preliminary results from Tenebrio molitor's feeding of different Enzyme fodders
(1) After feeding 30～40-day-old Tenebrio molitor by mixing the wheat bran and rice chaff, Alternanthera philoxeroides, corn stem, Trifolium L., rice stem, wheat bran in the ratio of 1:1, the result was obtained:the rice chaff could be the best choice on replacing wheat bran as feedstuff. After feeding for 30 days, the average fresh weighted 0.0246 g/larvae, the average fresh increasing by 0.0188 g/larvae, dry matter content for 39.98%, AD for 63.55%, ECD for 28.20%, and ECI for 17.93%.
(2) After feeding 40～50-day-old Tenebrio molitor by mixing the wheat bran and rice chaff in the quality ratio of 1:4,2:3,1:1,3:2 and 4:1, the result was obtained:the best choice is mixing the wheat bran and rice chaff in the ratio of 2:3 as feedstuff instead of the whole wheat fodder. After feeding for 30 days, the average fresh weighted 0.04915g/larvae, the average fresh quality increasing by 0.03329g/larvae, AD for 62.34%, ECD for 29.74%, and ECI for 19.50%.
(3) Using the two methods by pH test strips and pH-meter to determinate the pH value of Tenebrio molitor's intestines, the results were 4.3694 and 4.412 respectively. So the conclution is that the pH value of intestinal environment in Tenebrio molitor is about 4.4.
(4) Using the Enzyme fodders, which were joined with Cellulose, Hemicellulase and Xylanase respectively, to feed 50～60-day-old Tenebrio molitor, the conclution were as follows:①The 0.8% cellulose fodders could promote Tenebrio molitor's growth obviously After feeding for 30 days, the average fresh weighted 0.07015g/larvae, the average fresh quality increasing by 0.05549g/larvae, AD for 64.04%, ECD for 31:41%, and ECI for 26.72%.②The 1.0% Hemicellulase fodders could promote Tenebrio molitor's growth obviously. After feeding for 30 days, the average fresh weighted 0.06724g/larvae, the average fresh quality increasing by 0.05251g/larvae, AD for 61.60%, ECD for 44.78%, and ECI for 26.69%.③The 0.8% Xylanase fodders could promote Tenebrio molitor's growth obviously. After feeding for 30 days, the average fresh weighted 0.06595g/larvae, the average fresh quality increasing by 0.05133g/larvae, AD for 63.24%, ECD for 37.88%, and ECI for 23.95%.
2. The influence of edible fungi's growth from Tenebrio molitor's feces mediums
(1) The optimal preparation method of Tenebrio molitor's feces leaching solution through the test is as follows:leaching 16h on the condition of 70 degrees after mixing the feces and water by the ratio of 25:100.
(2) Cultivating Pleurotus ostreatus(Fr.)Kummer, FlammuLina velatiper(Fr.) Sing, Ganoderma lucidum (W.curt.:Fr.) Karst and Lentinus edodes (Berk.)Sing by use of solid and liquid mediums from Tenebrio molitof s feces leaching solution, the results show that:the hypha growth of the four mentioned mushrooms was the best in the flat mediums with Tenebrio molitor's feces leaching solution respectively being 25%,20%,25%,20%. In the liquid medium that used Tenebrio molitor's feces leaching solution as nitrogen source, the hypha growth of FlammuLina velatiper(Fr.) Sing, Ganoderma lucidum (W.curt.:Fr.) Karst and Lentinus edodes (Berk.)Sing was optimal with Tenebrio molitor's feces leaching solution being 20%; While the hypha growth of Pleurotus ostreatus(Fr.)Kummer was optimal with 25%.
(3) The result of using Tenebrio molitor's feces to cultivate and product edible fungis shows that:the Tenebrio molitor's feces could cultivate Lentinus edodes (Berk.)Sing, Pleurotus ostreatus(Fr.)Kummer, Flammulina velatiper(Fr.) Sing, Agrocybecha Sing Hnang, Copyinds comatus(MUⅡ.Fr) Gray and Ganoderma lucidum(W.curt.:Fr.) Karst; The content of feces is higher, the Flammulina velatiper (Fr.) Sing hypha growth faster; The production of Flammulina velatiper (Fr.) Sing and Pleurotus ostreatus(Fr.)Kummer could increase with the Tenebrio molitor's feces.
3.Preliminary studies on the methods of P-molting hormone extraction in Tenebrio molitor
This test used analysis method of Box-Behnken to analysis the effect on the extraction rate of P-molting hormone in Tenebrio molitor from interactions between time and temperature, and establish regression model table. The optimal Leaching condition was obtained:leaching 3 hours with ethanol on the condition of 80 degrees, with the extraction rate being 0.0563%. Besides, this test also obtained the optimal leaching ratio between material and liquid is 30:100, with the extraction rate being 0.0848%.
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