CN104208239B - 一种用于防治家禽大肠杆菌疾病的中药组合物、微生态制剂及制备方法 - Google Patents
一种用于防治家禽大肠杆菌疾病的中药组合物、微生态制剂及制备方法 Download PDFInfo
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Abstract
本发明公开了一种用于防治家禽大肠杆菌疾病的中药组合物、微生态制剂及制备方法，属于兽用中医药技术领域。该微生态制剂主要由以下原料制备而成：原料药以重量份数计为：黄连10～20份、黄柏10～20份、大黄5～10份、黄芪15～20份、板蓝根5～20份、甘草2～10份，菌液以体积份数计为：嗜酸乳杆菌菌液1～3份、粪肠球菌菌液1～6份、枯草芽孢杆菌菌液1～6份，菌液的总重量与原料药的总重量的比值为（1～3）:（40～50）。本发明采用纯天然中草药与三种益生菌复配制备微生态制剂，具有清热泻火、消炎止痢的功效，不仅对鸡肠道内致病性细菌引起的疾病具有良好的防治功效，还能促进动物机体的生长。
Description
技术领域
本发明具体涉及一种用于防治家禽大肠杆菌疾病的中药组合物，以及包含该中药组合物的微生态制剂及制备方法，属于兽用中医药技术领域。
背景技术
禽大肠杆菌病是由致病性大肠杆菌引起的疾病。致病性大肠杆菌在自然界中分布广泛，如鸡舍空气、灰尘、垫料、饲料、饮水、鸡体表以及孵化场、孵化器等用具中。该病一年四季均可发生，多雨、闷热、潮湿季节多发。各种日龄鸡均可感染，但主要侵害幼鸡和中鸡。病鸡和带菌鸡是主要的传染源，可经卵传递，鸡粪便污染种蛋是造成雏鸡感染最主要的传染来源，因污染的种蛋大肠杆菌可经蛋壳和壳膜侵入而造成鸡胚感染，也可经消化道和呼吸道感染。饲养管理不善，卫生状况差，营养不良以及感染其他疾病等都会诱发本病，由于禽大肠杆菌病易于与霉形体病合并感染，又常继发其他传染病如新城疫、禽流感惑、传染性支气管炎、巴氏杆菌病等，治疗十分困难，已成为危害养殖业的重要传染病，带来了巨大的经济损失。
目前，养殖户主要采用抗菌类药物对上述疾病进行治疗，但是抗生素类药物的滥用已使得众多耐药菌株频繁出现，治疗效果欠佳，且药物易在鸡体内及蛋类产品中残留，对人体危害极大。
发明内容
本发明的目的是提供一种用于防治家禽大肠杆菌疾病的中药组合物。
同时，本发明还提供一种用于防治家禽大肠杆菌疾病的微生态制剂。
最后，本发明还提供一种用于防治家禽大肠杆菌疾病的微生态制剂的制备方法。
为了实现以上目的，本发明所采用的技术方案是：
一种用于防治家禽大肠杆菌疾病的中药组合物，主要由以下重量份数的原料药组成：黄连10～20份、黄柏10～20份、大黄5～10份、黄芪15～20份、板蓝根5～20份、甘草2～10份。
优选的，一种用于防治家禽大肠杆菌疾病的中药组合物，主要由以下重量份数的原料药组成：黄连20份、黄柏20份、大黄10份、黄芪20份、板蓝根20份、甘草10份。
各原料药功效简述如下：
黄连：为毛茛科黄连属植物黄连、三角叶黄连或云南黄连的根茎；含有生物碱类如小檗碱、黄连碱、表小檗碱、黄柏酮等多种有效成分；性味苦、寒，归心、肝、胃、大肠经；能清热泻火、燥湿、解毒；主治热病邪入心经之高热、烦躁、谵妄或热盛迫血妄形之吐衄、湿热胸痞、泄泻、痢疾、心火亢盛之心烦失眠、胃热呕吐、消谷善饥、肝火目赤肿痛，以及热毒疮疡、疔毒走黄、牙龈肿痛、口舌生疮等；现代药理学证明黄连具有抗微生物、抗病原虫、抗病毒、抗心律失常、抗溃疡、抗肿瘤、抗炎等作用。酒黄连善清上焦火热，用于目赤、口疮；姜黄连清胃和胃止呕，用于寒热互结、湿热中阻、痞满呕吐；萸黄连舒肝和胃止呕，用于肝胃不和、呕吐吞酸。
黄柏：为芸香科黄檗属植物黄皮树或黄檗的树皮；含有生物碱类如小檗碱、黄柏碱、木兰花碱、药根碱等，还含有柠檬苦素、黄柏酮、菜油甾醇、豆甾醇等多种有效成分；性味苦、寒，归肾、膀胱、大肠经；能清热燥湿、泻火解毒；主治湿热痢疾、泄泻、黄疸、骨蒸劳热、口舌生疮、目赤肿痛、痈疽疮毒等；现代药理学证明黄柏具有抗病原微生物、解热和抗炎症、抗血小板聚集等功效，对心血管系统、消化系统和中枢肾经系统具有促进作用。
大黄：为蓼科大黄属植物掌叶大黄、唐古特大黄或药用大黄的根茎及根；含有游离蒽醌类如大黄酸、芦荟大黄素、大黄素等，综合蒽醌类如掌叶大黄素、大黄素甲醚-8-葡萄糖苷等，还含有双蒽醌类如掌叶大黄二蒽醌A、B、C，番泻苷A、B、C、D等，二苯乙烯苷类等多种活性成分；性味苦，寒，归胃、大肠、肝、脾经；能攻积滞，清湿热，泻火，凉血，祛瘀，解毒；主治实积便秘、热结胸痞，湿热泻痢、黄疸、淋病、水肿腹满、小便不利，目赤、咽喉肿痛、口舌生疮、胃热呕吐，吐血、咯血、衄血、便血、尿血，蓄血、经闭、产后瘀滞腹痛、癥瘕积聚、跌打损伤，热毒痈疡、丹毒、烫伤等；现代药理学证明大黄具有利胆、保肝、抗胃和十二指肠溃疡、抗病原微生物（细菌、真菌及病毒）、抗肿瘤、抗炎、止血、降脂等作用。
黄芪：为豆科黄芪属植物蒙古黄芪和膜荚黄芪的根；含有皂苷类成分如黄芪苷，异黄酮成分如毛蕊异黄酮，脂肪酸类如棕榈酸和亚油酸，多糖类如黄芪多糖、杂多糖等，以及近二十多种微量元素；黄芪性味甘，温，归肺、脾经；能益气升阳，固表止汗，利水消肿，托毒生肌；主治一切气虚血亏之症，如脾虚泄泻，肺虚咳嗽，脱肛，子宫下垂，自汗，盗汗，水肿，血痹，痈疽难溃或久溃不敛；现代药理学证明黄芪能影响机体代谢具有抗氧化作用，还能抗血栓、抗病毒、抗关节炎、抗癌、抗炎镇痛，保护心脑血管系统和促进造血功能。
板蓝根：为十字花科菘蓝属植物菘蓝的根；含有靛蓝、靛玉红及多种氨基酸，如精氨酸、谷氨酸等，还含有黑芥子苷、靛苷、色胺酮等多种有效成分；板蓝根性味苦，寒，归心、肺。肝、胃经；能清热，解毒，凉血，利咽；主治风热感冒，流感，流脑，乙脑，大头瘟疫，烂喉丹痧，单独等症；现代药理学证明板蓝根具有抗细菌、抗病毒、抗内毒素、抗癌等功效，同时对免疫系统具有调节作用。
甘草：为豆科甘草属植物甘草、光皮甘草、胀果甘草的根及根茎；含有三萜皂苷如甘草甜素、乌拉尔甘草皂苷A、B等，黄酮类化合物如甘草苷元、甘草苷等，香豆素类化合物如甘草香豆素、甘草酚等，生物碱类如喹啉类、异喹啉类等，多糖如甘草葡聚糖GBW、甘草多糖UA、UB、UC等，还含有甘草苯丙呋喃、正二十三烷等多种有效成分；甘草性味甘、平，归脾、胃、心、肺经；能和中缓急、润肺、解毒、调和诸药；炙用治脾胃虚弱、倦怠食少、腹痛便溏、四肢挛急疼痛、心悸、脏躁、肺痿咳嗽，生用治咽喉肿痛、痈疮肿毒、小儿胎毒及药物、食物中毒等；现代药理学证明甘草具有抗微生物、抗炎、镇咳、祛痰、抗肿瘤、抗突变、抗氧化等功效，对消化系统、心血管系统、生殖系统及脑、肾功能等具有调节作用。
一种用于防治家禽大肠杆菌疾病的中药组合物，可以采用常规方法制备成颗粒剂、粉剂、溶液剂、注射剂等常见兽药剂型。
一种用于防治家禽大肠杆菌疾病的微生态制剂，主要由以下原料制备而成：原料药以重量份数计为：黄连10～20份、黄柏10～20份、大黄5～10份、黄芪15～20份、板蓝根5～20份、甘草2～10份，菌液以体积份数计为：嗜酸乳杆菌菌液1～3份、粪肠球菌菌液1～6份、枯草芽孢杆菌菌液1～6份，菌液的总重量与原料药的总重量的比值为（1～3）:（40～50）。
优选的，一种用于防治家禽大肠杆菌疾病的微生态制剂，主要由以下原料制备而成：原料药以重量份数计为：黄连20份、黄柏20份、大黄10份、黄芪20份、板蓝根20份、甘草10份，菌液以体积份数计为：嗜酸乳杆菌菌液1份、粪肠球菌菌液5份、枯草芽孢杆菌菌液3份，菌液的总重量与原料药的总重量的比值为3:50。
所述的嗜酸乳杆菌菌液的浓度为0.5×108～1.0×108CFU/ml。
所述的粪肠球菌菌液的浓度为0.5×108～1.0×108CFU/ml。
所述的枯草芽孢杆菌菌液的浓度为0.5×108～1.0×108CFU/ml。
一种用于防治家禽大肠杆菌疾病的微生态制剂，可以采用常规方法制备。本发明仅提供一种具体的制备方法，包括以下步骤：
（1）按照重量份数准确取黄连、黄柏、大黄、黄芪、板蓝根和甘草，烘干后粉碎、过筛，再加水煎煮3次，合并3次煎液，滤过得药液备用；
（2）按照体积份数准确取嗜酸乳杆菌菌液、粪肠球菌菌液和枯草芽孢杆菌菌液，混匀后根据菌液总量与原料药总量之间的比值，将混合菌液加入到步骤（1）的药液中，在密闭条件下厌氧发酵，发酵产物经冷冻干燥即得。
所述步骤（1）中的烘干温度为60～80℃。
所述步骤（1）中的过筛为过60～100目筛。
所述步骤（1）中加水煎煮为每次煎煮加5～6倍量的水，煎煮1～2小时。
所述步骤（2）中嗜酸乳杆菌菌液的制备方法为：先将嗜酸乳杆菌接种于斜面培养基中，将斜面培养基上产生的悬浮液加入到MRS液体培养基中扩大培养，至对数生长期时菌液的浓度为0.5×108～1.0×108CFU/ml即得。
所述步骤（2）中粪肠球菌菌液的制备方法为：先将粪肠球菌接种于斜面培养基中，将斜面培养基上产生的悬浮液加入到MRS液体培养基中扩大培养，至对数生长期时菌液的浓度为0.5×108～1.0×108CFU/ml即得。
所述步骤（2）中枯草芽孢杆菌菌液的制备方法为：先将枯草芽孢杆菌接种于斜面培养基中，将斜面培养基上产生的悬浮液加入到LB液体培养基中扩大培养，至对数生长期时菌液的浓度为0.5×108～1.0×108CFU/ml即得。
所述步骤（2）中厌氧发酵的温度为30～37℃，发酵时间为36～72小时。
微生态制剂的用法与用量：混饮，每100g兑水200kg，连用2～3天，为预防剂量；治疗量加倍。
本发明的有益效果：
本发明是在中兽医学卫气营血辩证理论方法的指导下，筛选出黄连、黄柏、大黄、黄芪、板蓝根、甘草共6味纯天然中草药，按照君臣佐使原则配伍组方。方中大黄清热泻火、攻击导滞、凉血解毒，为君药；黄连清醒，黄芩清心火、胃火、肺火，黄柏清下焦之火，辅以黄连、黄芩、黄柏苦寒燥湿，以解肠中热毒；板蓝根清热解毒，甘草缓急、止痛，共为佐使。诸药合用共奏清热解毒、抗菌及增强机体免疫力之功效；主要用于防治家禽大肠杆菌疾病，见效快，疗效确切。
本发明采用嗜酸乳杆菌、粪肠球菌和枯草芽孢杆菌制备微生态制剂，其中嗜酸乳杆菌能调整肠道菌群平衡，抑制肠道不良微生物的增殖，分泌抗生物素类物质如嗜酸乳菌素、嗜酸杆菌素、乳酸菌素等，对肠道致病菌产生拮抗作用；粪肠球菌有利于改善肠道内微生态平衡，防治动物肠道菌丛区系紊乱，还能分解蛋白质为小肽，合成B族维生素，增强巨嗜细胞的活性，促进动物的免疫反应，提高抗体水平等；枯草芽孢杆菌能迅速消耗环境中的游离氧，造成肠道低氧，促进有益厌氧菌生长，并产生乳酸等有机酸类，降低肠道PH值，间接抑制其它致病菌生长。刺激动物免疫器官的生长发育，激活T、B淋巴细胞，提高免疫球蛋白和抗体水平，增强细胞免疫和体液免疫功能，提高群体免疫力。三种益生菌联用可通过动物消化道生物的竞争性排斥作用，抑制病原菌繁殖，阻止病原菌吸附于肠壁，促进其排出体外；补充有益菌群，调整动物肠道菌群微生态平衡；发挥营养助消化作用，提高饲料转化率，合成营养物质，促进动物生长；还可通过增强非特异性免疫功能来预防疾病，提高机体免疫力；抑制肠道内氨及胺等毒性物质的产生，改善环境卫生，净化环境。
本发明采用纯天然中草药，原料药易得，价格低廉，安全、无有害残留，对畜禽机体的毒副作用小，不易产生抗药性，主要通过整体作用、双向调节、组合效应、扶正祛邪发挥药物功效。与三种益生菌复配制备微生态制剂，具有清热泻火、消炎止痢的功效，不仅对鸡肠道内致病性细菌引起的疾病具有良好的防治功效，还能促进动物机体的生长，有效控制鸡大肠杆菌病，达到标本兼制的目的。
本发明制备微生态制剂的方法简单、易操作，使用方便，成本低，适于大规模生产及推广应用。
具体实施方式
下述实施例仅对本发明作进一步详细说明，但不构成对本发明的任何限制。
实施例1
本实施例中用于防治家禽大肠杆菌疾病的中药组合物，由以下重量份数的原料药组成：黄连10份、黄柏20份、大黄5份、黄芪20份、板蓝根5份、甘草10份。制备方法为：按照重量份数准确取各味原料药，粉碎后混匀、过60目筛，以250g/袋分装即得。
用法与用量：混饲，每100kg饲料拌药1kg，自由采食，连用3～5天。
实施例2
本实施例中用于防治家禽大肠杆菌疾病的中药组合物，由以下重量份数的原料药组成：黄连20份、黄柏20份、大黄10份、黄芪20份、板蓝根20份、甘草10份。制备方法为：按照重量份数准确取各味原料药，粉碎后混匀、过60目筛，以250g/袋分装即得。
用法与用量：混饲，每100kg饲料拌药1kg，自由采食，连用3～5天。
实施例3
本实施例中用于防治家禽大肠杆菌疾病的微生态制剂，由以下原料制备而成：原料药以重量份数计为：黄连10份、黄柏20份、大黄5份、黄芪20份、板蓝根5份、甘草10份，菌液以体积份数计为：嗜酸乳杆菌菌液1份、粪肠球菌菌液6份、枯草芽孢杆菌菌液1份，菌液的总重量与原料药的总重量的比值为3:50。
本实施例中用于防治家禽大肠杆菌疾病的微生态制剂的制备方法，包括以下步骤：
（1）按照重量份数准确取黄连、黄柏、大黄、黄芪、板蓝根和甘草，60℃烘干后粉碎，过60目筛，再加水煎煮3次，每次煎煮加5倍量的水，煎煮1.5小时，合并3次煎液，滤过得药液备用；
（2）按照体积份数准确取嗜酸乳杆菌菌液、粪肠球菌菌液和枯草芽孢杆菌菌液，混匀后根据菌液总量与原料药总量之间的比值，将混合菌液加入到步骤（1）的药液中，在密闭条件下厌氧发酵，发酵的温度为30℃，发酵时间为72小时，将发酵产物冷冻干燥即得。
所述步骤（2）中嗜酸乳杆菌菌液、粪肠球菌菌液、枯草芽孢杆菌菌液的制备方法为：先将嗜酸乳杆菌、粪肠球菌、粪肠球菌分别接种于斜面培养基中，将各个斜面培养基上产生的悬浮液依次加入到MRS液体培养基、MRS液体培养基、LB液体培养基中扩大培养，至对数生长期时菌液的浓度分别为0.5×108～1.0×108CFU/ml即得。
用法与用量：混饮，每100g兑水200kg，连用2～3天，为预防剂量；治疗量加倍。
实施例4
本实施例中用于防治家禽大肠杆菌疾病的微生态制剂，由以下原料制备而成：原料药以重量份数计为：黄连20份、黄柏20份、大黄10份、黄芪20份、板蓝根20份、甘草10份，菌液以体积份数计为：嗜酸乳杆菌菌液1份、粪肠球菌菌液5份、枯草芽孢杆菌菌液3份，菌液的总重量与原料药的总重量的比值为1:40。
本实施例中用于防治家禽大肠杆菌疾病的微生态制剂的制备方法，包括以下步骤：
（1）按照重量份数准确取黄连、黄柏、大黄、黄芪、板蓝根和甘草，80℃烘干后粉碎，过100目筛，再加水煎煮3次，每次煎煮加6倍量的水，煎煮2小时，合并3次煎液，滤过得药液备用；
（2）按照体积份数准确取嗜酸乳杆菌菌液、粪肠球菌菌液和枯草芽孢杆菌菌液，混匀后根据菌液总量与原料药总量之间的比值，将混合菌液加入到步骤（1）的药液中，在密闭条件下厌氧发酵，发酵的温度为37℃，发酵时间为36小时，将发酵产物冷冻干燥即得。
所述嗜酸乳杆菌菌液、粪肠球菌菌液、枯草芽孢杆菌菌液的制备方法同上。
用法与用量：混饮，每100g兑水200kg，连用2～3天，为预防剂量；治疗量加倍。
实施例5
本实施例中用于防治家禽大肠杆菌疾病的微生态制剂，由以下原料制备而成：原料药以重量份数计为：黄连20份、黄柏10份、大黄10份、黄芪15份、板蓝根20份、甘草2份，菌液以体积份数计为：嗜酸乳杆菌菌液3份、粪肠球菌菌液1份、枯草芽孢杆菌菌液6份，菌液的总重量与原料药的总重量的比值为2:45。
本实施例中用于防治家禽大肠杆菌疾病的微生态制剂的制备方法，包括以下步骤：
（1）按照重量份数准确取黄连、黄柏、大黄、黄芪、板蓝根和甘草，70℃烘干后粉碎，过80目筛，再加水煎煮3次，每次煎煮加5倍量的水，煎煮1.5小时，合并3次煎液，滤过得药液备用；
（2）按照体积份数准确取嗜酸乳杆菌菌液、粪肠球菌菌液和枯草芽孢杆菌菌液，混匀后根据菌液总量与原料药总量之间的比值，将混合菌液加入到步骤（1）的药液中，在密闭条件下厌氧发酵，发酵的温度为35℃，发酵时间为50小时，将发酵产物冷冻干燥即得。
所述嗜酸乳杆菌菌液、粪肠球菌菌液、枯草芽孢杆菌菌液的制备方法同上。
用法与用量：混饮，每100g兑水200kg，连用2～3天，为预防剂量；治疗量加倍。
试验例
试验动物：从养殖场选择1000只鸡，进行大肠杆菌攻毒，待鸡群发病后进行临床疗效验证。
试验药物：实施例3～5制备的中草药微生态制剂以及对照市售中药。
试验方法：从患病鸡群中随机选取500只确诊病鸡，分为5组；试验组1～3依次给予实施例3～5制备的微生态制剂；对照组给予对照组药物；空白对照组不给于任何药物。试验组1～3及对照组中药物的用法用量为：混饮，每100g兑水100kg，连用7天。患鸡正常采食和饮水，每天观察患鸡的临床变化，统计治愈、有效及无效数，计算治愈率和总有效率，试验结果详见下表1。
疗效判断标准：
治愈：试验结束后，病鸡症状消失，精神转好，食欲增长，完全恢复正常；
有效：试验结束后，鸡病症状、精神、食欲明显好转；
无效：试验期间出现症状，试验结束后仍不见症状消失。
表1各处理组对鸡大肠杆菌感染的治疗效果
结论：从表1可以看出，试验组1～3的治愈率达80%以上，总有效率达95%以上；而对照组的总有效率仅为82%，治愈率仅为70%。统计分析表明，试验组的治愈率及总有效率显著高于对照组（P＜0.01）。
试验结果表明，本发明实施例制备的微生态制剂对鸡大肠杆菌疾病具有良好的防治功效。整个用药期间试验组鸡群未发现任何异常，表明本发明实施例制备的散剂及微生态制剂的毒副作用小。
Claims (8)
1.一种用于防治家禽大肠杆菌疾病的微生态制剂，其特征在于：由以下原料制备而成：原料药以重量份数计为：黄连10～20份、黄柏10～20份、大黄5～10份、黄芪15～20份、板蓝根5～20份、甘草2～10份，菌液以体积份数计为：嗜酸乳杆菌菌液1～3份、粪肠球菌菌液1～6份、枯草芽孢杆菌菌液1～6份，上述三种菌液的浓度均为0.5×108～1.0×108CFU/ml；菌液的总重量与原料药的总重量的比值为(1～3):(40～50)；该微生态制剂是由包括以下步骤的方法制备的：
(1)按照重量份数准确取黄连、黄柏、大黄、黄芪、板蓝根和甘草，烘干后粉碎、过筛，再加水煎煮3次，合并3次煎液，滤过得药液备用；
(2)按照体积份数准确取嗜酸乳杆菌菌液、粪肠球菌菌液和枯草芽孢杆菌菌液，混匀后根据菌液总量与原料药总量之间的比值，将混合菌液加入到步骤(1)的药液中，在密闭条件下厌氧发酵，发酵产物经冷冻干燥即得。
2.根据权利要求1所述的用于防治家禽大肠杆菌疾病的微生态制剂，其特征在于：由以下原料制备而成：原料药以重量份数计为：黄连20份、黄柏20份、大黄10份、黄芪20份、板蓝根20份、甘草10份，菌液以体积份数计为：嗜酸乳杆菌菌液1份、粪肠球菌菌液5份、枯草芽孢杆菌菌液3份，菌液的总重量与原料药的总重量的比值为3:50。
3.一种如权利要求1或2所述的用于防治家禽大肠杆菌疾病的微生态制剂的制备方法，其特征在于：包括以下步骤：
(1)按照重量份数准确取黄连、黄柏、大黄、黄芪、板蓝根和甘草，烘干后粉碎、过筛，再加水煎煮3次，合并3次煎液，滤过得药液备用；
(2)按照体积份数准确取嗜酸乳杆菌菌液、粪肠球菌菌液和枯草芽孢杆菌菌液，混匀后根据菌液总量与原料药总量之间的比值，将混合菌液加入到步骤(1)的药液中，在密闭条件下厌氧发酵，发酵产物经冷冻干燥即得。
4.根据权利要求3所述的用于防治家禽大肠杆菌疾病的微生态制剂的制备方法，其特征在于：所述步骤(1)中加水煎煮为每次煎煮加5～6倍量的水，煎煮1～2小时。
5.根据权利要求3所述的用于防治家禽大肠杆菌疾病的微生态制剂的制备方法，其特征在于：所述步骤(2)中嗜酸乳杆菌菌液的制备方法为：先将嗜酸乳杆菌接种于斜面培养基中，将斜面培养基上产生的悬浮液加入到MRS液体培养基中扩大培养，至对数生长期时菌液的浓度为0.5×108～1.0×108CFU/ml即得。
6.根据权利要求3所述的用于防治家禽大肠杆菌疾病的微生态制剂的制备方法，其特征在于：所述步骤(2)中粪肠球菌菌液的制备方法为：先将粪肠球菌接种于斜面培养基中，将斜面培养基上产生的悬浮液加入到MRS液体培养基中扩大培养，至对数生长期时菌液的浓度为0.5×108～1.0×108CFU/ml即得。
7.根据权利要求3所述的用于防治家禽大肠杆菌疾病的微生态制剂的制备方法，其特征在于：所述步骤(2)中枯草芽孢杆菌菌液的制备方法为：先将枯草芽孢杆菌接种于斜面培养基中，将斜面培养基上产生的悬浮液加入到LB液体培养基中扩大培养，至对数生长期时菌液的浓度为0.5×108～1.0×108CFU/ml即得。
8.根据权利要求3所述的用于防治家禽大肠杆菌疾病的微生态制剂的制备方法，其特征在于：所述步骤(2)中厌氧发酵的温度为30～37℃，发酵时间为36～72小时。
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