CN116849293A - 一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用 - Google Patents
一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用 Download PDFInfo
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Abstract
本发明公开了一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用,共分为菌种活化与发酵、菌液混合、添加剂加入、凝胶化处理、菌剂二次加工五个步骤;本发明在制备菌剂的流程时刻保持凝胶状复合菌剂内部复合菌群的活性,所得凝胶易于随时取用,且可以根据实际情况进行切割、挤压,增大与饲料的接触面积,提高家蚕食用率;将传统液态菌剂改变为凝胶状,改善了传统喷洒菌剂后造成的一定时间内蚕座湿度提升、家蚕生存环境改变而导致的生病、饲料变质问题。
Description
技术领域
本发明涉及家蚕饲料添加剂技术领域,尤其涉及一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用及应用。
背景技术
家蚕的英文名为“silkworm”(意为“丝虫”)是因为它用丝织茧。茧是由一根300-900米长的丝织成的。现如今我国茧丝绸产量与出口量均占世界总量的70%以上,已成为可以主导世界茧丝价格走势的茧丝绸大国。蚕是变态类昆虫,最常见的是桑蚕,又称家蚕,以桑叶为食料的吐丝结茧的经济昆虫之一。桑蚕起源于中国,其发育温度是7~40℃,饲育适温为20~30℃。幼蚕以桑叶为生,不断吃桑叶后身体便成白色,一段时间后它便开始脱皮,脱皮持续约一天的时间。经过一次脱皮后,就是二龄幼虫,幼虫共要脱皮四次,成为五龄幼虫,再吃桑叶8天成为熟蚕,开始吐丝结茧。随着环境污染、劳动力成本的提高以及现代农业的发展,人工饲料养蚕已经成为蚕桑业新的发展趋势。但是目前,人工饲料育的家蚕体质量、抗病能力及蚕茧产量和质量与桑叶育家蚕均存在一定差距,大大影响了家蚕人工饲料的产业化进程。最近研究发现,人工饲料育家蚕与桑叶育家蚕肠道微生物菌群结构存在明显差异,因此改善人工饲料育家蚕肠道菌群有助于提高家蚕抗病性、蚕茧等级、产丝量及蚕丝品质等。
另外在人工饲料养蚕过程中对环境温湿度的要求较高,因此对湿饲料喷洒菌液后,必须要等待饲料表面水分稍干后,才可转移至蚕箱进行饲喂,湿度过高影响家蚕周围环境,进而可能会造成家蚕生病甚至死亡等情况出现;为了保证微生物菌剂与人工饲料正常混合使用,在不影响环境湿度的同时保证饲喂进度,改善微生物菌剂形态是一种较好的方法;将微生物菌剂凝胶化,即可以保证饲喂过程中与饲料均匀混合,同时也可减少湿度过高对家蚕生长造成的影响。
因此,本领域的技术人员致力于开发一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用,以解决上述现有技术的不足。
发明内容
有鉴于现有技术的上述缺陷,本发明所要解决的技术问题是目前人工饲料养蚕蚕体抗病性差、饲育成绩较低以及常规液体菌剂喷洒在人工饲料上后易造成蚕箱湿度迅速升高,饲料易变质及家蚕受到湿度变化影响导致生病甚至死亡的缺陷问题。
为实现上述目的,本发明第一方面提供了一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用,所述方法包括以下步骤:
步骤1、准备菌种,菌种单独活化与发酵,扩大培养;
步骤2、混合扩大培养后的菌液,获得混合菌液;
步骤3、在混合菌液中添加补充剂、稳定剂,混合后获得复合混合菌液;
步骤4、发酵菌液加入增稠剂,凝胶化处理后获得凝胶状微生物复合菌剂;
步骤5、凝胶状微生物复合菌剂根据养蚕箱尺寸进行二次处理后,得到可直接使用的成品;
对于步骤1,所述菌种包括枯草芽孢杆菌、粪肠球菌、光合细菌、双歧杆菌;所述光合细菌为沼泽红假单胞菌;所述活化与发酵过程包括制备培养液、菌种接种、控制温度发酵;
对于步骤2,所述混合菌液中,经活化扩大培养后的枯草芽孢杆菌、粪肠球菌、光合细菌、双歧杆菌相应菌液添加体积比为5~8:0.5~0.8:0.5~1:0.5~1.5;
对于步骤3,所述补充剂包括葡萄糖、复合维生素粉、复合氨基酸粉、混合无机盐、喹乙醇;所述补充剂中各成分的重量百分比为葡萄糖40%~55%、复合维生素粉13%~25%、复合氨基酸粉15%~18%、混合无机盐3%~8%、喹乙醇2%~5%;所述稳定剂柠檬酸、乙氧基喹啉、二丁基羟基甲苯、EDTA、茶多酚包括中的一种或多种;所述补充剂、稳定剂占复合混合菌液重量的5%~8%;
对于步骤4,所述增稠剂包括明胶、琼脂、果胶、海藻酸钠、黄原胶、魔芋胶中的一种或多种;所述凝胶化处理包括调节混合环境、加入复合混合菌液、静置调温获得凝胶状微生物复合菌剂;
对于步骤5,所述二次处理包括切块、称重、包装;
本发明第二方面提供了一种家蚕人工饲料育微生物复合菌剂在家蚕饲养方向上的应用,本发明的凝胶化微生物复合菌剂在使用时即能将其置于机器中从挤出,平铺至已放入养蚕箱的人工饲料上,也能直接放入块状的成品凝胶状微生物复合菌剂,与人工饲料一同饲喂家蚕;
采用以上方案,本发明公开的一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用,具有以下优点:
(1)本发明的一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用,将传统液态菌剂改变为凝胶状,改善了传统喷洒菌剂后造成的一定时间内蚕箱湿度提升、家蚕生存环境改变而导致的生病、饲料变质问题。
(2)本发明的一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用,制备流程时刻保持凝胶状复合菌剂内部复合菌群的活性,所得凝胶易于随时取用,且可以根据实际情况进行切割、挤压,增大与饲料的接触面积,提高家蚕食用率。
综上所述,本发明公开的一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用,制备流程时刻保持凝胶状复合菌剂内部复合菌群的活性,所得凝胶易于随时取用,且可以根据实际情况进行切割、挤压,增大与饲料的接触面积,提高家蚕食用率;将传统液态菌剂改变为凝胶状,改善了传统喷洒菌剂后造成的一定时间内蚕箱湿度提升、家蚕生存环境改变而导致的生病、饲料变质问题。
以下将结合具体实施方式对本发明的构思、具体技术方案及产生的技术效果作进一步说明,以充分地了解本发明的目的、特征和效果。
附图说明
图1为本发明一种专用于家蚕人工饲料育微生物复合菌剂的制备步骤示意图。
具体实施方式
以下介绍本发明的多个优选实施例,使其技术内容更加清楚和便于理解。本发明可以通过许多不同形式的实施例来得以体现,这些实施例为示例性描述,本发明的保护范围并非仅限于文中提到的实施例。
如若有未注明具体条件的实验方法,通常按照常规条件,如相关说明书或者手册进行实施。
如图1所示,本发明的家蚕人工饲料育专用微生物复合菌剂的制备方法及应用,实施例、家蚕人工饲料育专用微生物复合菌剂的制备方法及应用
步骤1、准备菌种,菌种单独活化与发酵,扩大培养;
枯草芽孢杆菌的活化与培养过程为:将保藏的枯草芽孢杆菌接种到LB培养基中,温度设置为37℃,pH为7,摇床振荡培养20~28h;培养好后接种到灭菌好的发酵罐中,接种量为2%,加入营养液,温度控制为37℃左右,搅拌转速为200r/min,时间持续1h~2h,后续转速调整为300~350r/min,时间持续20~24h;在机器启动过程中,每隔1~2h记录一次pH值,记录转速、菌液温度及菌浓;在查看前后都应以高温蒸汽对取样口灭菌,以免杂菌污染菌液;
粪肠球菌活化与培养过程为:取粪肠球菌冻干菌溶解于液体培养基中活化菌种,将活化后的菌种解冻后接入种子培养基中,37℃,150r/min条件下培养18h;培养好后接种到灭菌好的发酵罐中,接种量为3%,加入营养液,温度控制为37℃,pH为6.2,搅拌转速为100r/min,在机器启动过程中,每隔1~2h记录一次pH值,记录转速、菌液温度及菌浓;在查看前后都应以高温蒸汽对取样口灭菌,以免杂菌污染菌液;
沼泽红假单胞菌的活化与培养过程为:取沼泽红假单胞菌冻干菌溶解于液体培养基中活化菌种,将活化后的菌种置于光照强度为2500勒克斯的光照培养箱中,保持温度28~31℃,pH为5.5-7,持续24h进行扩大培养;将扩大培养后的活化菌液接入培养罐中,接种量为6.5%,保持同样温度与光照强度,持续6~8h;每隔1~2h记录一次pH值,记录转速、菌液温度及菌浓;在查看前后都应以高温蒸汽对取样口灭菌,以免杂菌污染菌液;
双歧杆菌的活化与培养过程为:取双歧杆菌冻干粉溶解于液体培养基中活化,后续接入发酵罐中,接种量为5%,设置温度37~42℃,pH为6.5~7.0,发酵培养10~15h;每隔1~2h记录一次pH值,记录转速、菌液温度及菌浓;在查看前后都应以高温蒸汽对取样口灭菌,以免杂菌污染菌液;
步骤2、混合扩大培养后的菌液,获得混合菌液;
经活化扩大培养后的枯草芽孢杆菌、粪肠球菌、光合细菌、双歧杆菌相应菌液添加体积比为5~8:0.5~0.8:0.5~1:0.5~1.5,按照耐受pH值由小至大的顺序进行添加混合,依次为沼泽红假单胞菌菌液、粪肠球菌菌液、双歧杆菌菌液、枯草芽孢杆菌菌液;保持温度在30~33℃,pH为6~6.5,搅拌器混合均匀后备用;
步骤3、在混合菌液中添加补充剂、稳定剂,混合后获得复合混合菌液;
补充剂中各成分的重量百分比为葡萄糖40%~55%、复合维生素粉(维生素A、维生素E、维生素B1、维生素B2、维生素E)13%~25%、复合氨基酸粉(甘氨酸、苏氨酸)15%~18%、混合无机盐3%~8%、喹乙醇2%~5%;本实施例选用的稳定剂为柠檬酸及乙氧基喹啉;补充剂、稳定剂占复合混合菌液重量的5%~8%;
步骤4、发酵菌液加入增稠剂,凝胶化处理后获得凝胶状微生物复合菌剂;
本实施例选用的增稠剂为黄原胶及魔芋胶,复配共混胶浓度为1%~1.2%,将黄原胶粉与魔芋胶粉混合,按批次缓慢加入复合混合菌液,保持温度在30~35℃;在混合过程中根据pH值变化适当加入醋酸溶液,使混合菌液整体呈中性至弱酸性;所有复合混合菌液全部倒入完毕后,持续搅拌10~20min,转速为80~100r/min;搅拌均匀后倒入平盘中;
步骤5、凝胶状微生物复合菌剂二次处理后,得到可直接使用的成品;
将平盘中的增稠剂与复合混合菌液的混合溶液置于室温或10℃冷藏库中静置,硬度达到10.5×103~17.5×103N/m2后切块,切块尺寸与养蚕箱尺寸适应,呈条状或方块状;称重后按照每次饲喂量进行初包装,得到可直接使用的凝胶状微生物复合菌剂;
实际投入使用后,可直接将本凝胶状微生物复合菌剂均匀铺在人工饲料上,如有配套机器,可将本发明的凝胶状微生物复合菌剂置于压力机器中,并挤压呈丝状,使其与饲料的接触面积更大;经试验,在饲喂过程中蚕箱湿度几乎无变化,桑叶及饲料未出现因湿度过高而导致的变质情况出现,家蚕未出现死亡情况;家蚕吐丝成茧后,蚕茧产量单重增加9.8%,较往年同期的疾病死亡率降低23.7%;
将实施例进行分析可知,本发明的一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用制备流程时刻保持凝胶状复合菌剂内部复合菌群的活性,所得凝胶易于随时取用,且可以根据实际情况进行切割、挤压,增大与饲料的接触面积,提高家蚕食用率;将传统液态菌剂改变为凝胶状,改善了传统喷洒菌剂后造成的一定时间内蚕箱湿度提升、家蚕生存环境改变而导致的生病、饲料变质问题。
以上详细描述了本发明的较佳具体实施例。应当理解,本领域的普通技术无需创造性劳动就可以根据本发明的构思做出诸多修改和变化。因此,凡本技术领域中技术人员依本发明的构思在现有技术的基础上通过逻辑分析、推理或者有限的试验可以得到的技术方案,皆应在由权利要求书所确定的保护范围内。
Claims (6)
1.一种家蚕人工饲料育专用微生物复合菌剂的制备方法及应用,其特征在于,
所述方法包括以下步骤:
步骤1、准备菌种,菌种单独活化与发酵,扩大培养;
步骤2、混合扩大培养后的菌液,获得混合菌液;
步骤3、在混合菌液中添加补充剂、稳定剂,混合后获得复合混合菌液;
步骤4、发酵菌液加入增稠剂,凝胶化处理后获得凝胶状微生物复合菌剂;
步骤5、凝胶状微生物复合菌剂根据养蚕匾尺寸进行二次处理后,得到可直接使用的成品;
对于步骤1,所述活化与发酵过程包括制备培养液、菌种接种、控制温度发酵;
对于步骤4,所述凝胶化处理包括调节混合环境、加入复合混合菌液、静置调温获得凝胶状微生物复合菌剂;
对于步骤5,所述二次处理包括切块、称重、包装。
2.如权利要求1所述家蚕人工饲料育专用微生物复合菌剂的制备方法,其特征在于,
对于步骤1,所述菌种包括枯草芽孢杆菌、粪肠球菌、光合细菌、双歧杆菌;所述光合细菌为沼泽红假单胞菌。
3.如权利要求1所述家蚕人工饲料育专用微生物复合菌剂的制备方法,其特征在于,
对于步骤2,所述混合菌液中,经活化扩大培养后的枯草芽孢杆菌、粪肠球菌、光合细菌、双歧杆菌相应菌液添加体积比为5~8:0.5~0.8:0.5~1:0.5~1.5。
4.如权利要求1所述家蚕人工饲料育专用微生物复合菌剂的制备方法,其特征在于,
对于步骤3,所述补充剂包括葡萄糖、复合维生素粉、复合氨基酸粉、混合无机盐、喹乙醇;所述补充剂中各成分的重量百分比为葡萄糖40%~55%、复合维生素粉13%~25%、复合氨基酸粉15%~18%、混合无机盐3%~8%、喹乙醇2%~5%;所述稳定剂柠檬酸、乙氧基喹啉、二丁基羟基甲苯、EDTA、茶多酚包括中的一种或多种;所述补充剂、稳定剂占复合混合菌液重量的5%~8%。
5.如权利要求1所述家蚕人工饲料育专用微生物复合菌剂的制备方法,其特征在于,
所述增稠剂包括明胶、琼脂、果胶、海藻酸钠、黄原胶、魔芋胶中的一种或多种。
6.一种家蚕人工饲料育微生物复合菌剂在家蚕饲养方向上的应用,其特征在于,
本发明的凝胶化微生物复合菌剂在使用时将其置于机器中从挤出,平铺至已放入养蚕箱的人工饲料上,或直接放入块状的成品凝胶状微生物复合菌剂,与人工饲料一同饲喂家蚕,最终饲料变质情况、蚕茧单重及疾病死亡率都有改善。
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