CN106259069A - 一种嗜酸乳杆菌降解动物体内真菌毒素的方法 - Google Patents
一种嗜酸乳杆菌降解动物体内真菌毒素的方法 Download PDFInfo
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Abstract
本发明属于毒素降解技术领域，公开了嗜酸乳杆菌在降解动物体内真菌毒素中的应用，所述真菌毒素为T‑2毒素、黄曲霉毒素B1、呕吐毒素和赭曲霉毒素；将嗜酸乳杆菌菌液均匀喷洒至动物饲料中，经过十天投喂含有乳酸菌饲料投喂含有T‑2毒素、（AFB1）、呕吐毒素（DON）、赭曲霉（OTA）毒素的南美白对虾和罗非鱼，对虾和罗非鱼肌肉真菌毒素残留较对照组有显著下降，其中对虾肌肉T‑2下降63.86±1.32，AFB1下降53.04±0.97%，DON下降70.74±1.47%，OTA下降70.24±1.22%。罗非鱼肌肉T‑2下降54.93±2.52%，AFB1下降42.18±1.97%，DON下降48.75±0.93%，OTA下降32.79±1.81%；本发明将嗜酸乳杆菌制成饲料添加剂，即可实现降解动物体内累积的真菌毒素，具有实际的应用价值。
Description
技术领域
本发明涉及毒素降解技术领域，更具体地，涉及一种利用嗜酸乳杆菌降解动物体内真菌毒素的方法。
背景技术
真菌毒素对人和动物具有毒性作用，目前已知的真菌毒素已有200种以上，其中有相当部分具有较强的致突变、致癌作用和致畸性。已知产毒真菌主要有：曲霉菌属、青霉菌属、镰刀菌属等。T-2毒素、黄曲霉毒素B1（AFB1）、呕吐毒素（DON）和赭曲霉毒素（OTA）是霉菌生长产生的有毒代谢产物，属对虾饲料和鱼饲料中自然发生的常见真菌毒素，毒性强，不易清除，性质稳定，可引起对虾和鱼疾病的发生以及生产性能下降、肌肉品质降低等，对水产业造成巨大经济损失，阻碍了水产业的发展，同时含有真菌毒素的鱼虾产品通过食物链的传递直接或间接危害人类的生命健康安全，因此，有效控制其危害是迫在眉睫的热点问题。
乳酸菌作为益生菌，被广泛应用于各种产品应用中，目前有很多关于乳酸菌体外降解真菌毒素的报道，但单种菌降解毒素的种类有限，且效果不佳，另外不能在动物体内起到作用，不能起到降解动物体内累积的真菌毒素的作用。
发明内容
本发明所要解决的技术问题是克服现有技术存在的上述缺陷，提供一种利用嗜酸乳杆菌降解动物体内真菌毒素的方法。
本发明的目的是通过以下技术方案予以实现的：
嗜酸乳杆菌在降解动物体内真菌毒素中的应用，所述真菌毒素为T-2毒素、黄曲霉毒素B1、呕吐毒素和赭曲霉毒素。
本发明通过从酸奶中分离出嗜酸乳杆菌，其具有同时降解T-2毒素、黄曲霉毒素B1、呕吐毒素和赭曲霉毒素四种毒素的作用。
需要说明的是，发明人研究了多种市售酸奶，并从中分离出多株嗜酸乳杆菌，其均具有降解上述真菌毒素的作用。
本发明还提供一种利用嗜酸乳杆菌降解动物体内真菌毒素的方法，是将嗜酸乳杆菌菌液与动物饲料混合后饲喂动物。
具体的，所述混合可以是将嗜酸乳杆菌菌液均匀喷洒至动物饲料中。
优选地，所述方法中，所述嗜酸乳杆菌菌液的体积与动物饲料的质量比为1：1。
优选地，所述嗜酸乳杆菌菌液中菌含量为1.2×109～6×109CFU/mL。
优选地，所述动物为罗非鱼和/或对虾。
与现有技术相比，本发明具有以下有益效果：
本发明提供了嗜酸乳杆菌在降解动物体内真菌毒素中的应用，所述真菌毒素为T-2毒素、黄曲霉毒素B1、呕吐毒素和赭曲霉毒素；将嗜酸乳杆菌菌液均匀喷洒至动物饲料中，经过十天投喂含有乳酸菌饲料投喂含有T-2毒素、（AFB1）、呕吐毒素（DON）、赭曲霉（OTA）毒素的南美白对虾和罗非鱼，对虾和罗非鱼肌肉真菌毒素残留较对照组有显著下降，其中对虾肌肉T-2下降63.86±1.32，AFB1下降53.04±0.97%，DON下降70.74±1.47%，OTA下降70.24±1.22%。罗非鱼肌肉T-2下降54.93±2.52%，AFB1下降42.18±1.97%，DON下降48.75±0.93%，OTA下降32.79±1.81%；本发明将嗜酸乳杆菌制成饲料添加剂，即可实现降解动物体内累积的真菌毒素，具有实际的应用价值。
附图说明
图1为嗜酸乳杆菌的菌落形态。
图2为嗜酸乳杆菌镜检图。
图3为罗非鱼肌肉真菌毒素消减率与肠道乳酸菌量比的关系。
图4为对虾肌肉真菌毒素消减率与肠道乳酸菌量比的关系。
具体实施方式
下面结合说明书附图和具体实施例进一步说明本发明的内容，但不应理解为对本发明的限制。在不背离本发明精神和实质的情况下，对本发明方法、步骤或条件所作的简单修改或替换，均属于本发明的范围；若未特别指明，实施例中所用的技术手段为本领域技术人员所熟知的常规手段。
实施例1
（1）对虾和罗非鱼染毒试验
随机挑选体质健康、个体大小均一的南美白对虾（每尾5±0.5g），按照每组40尾（设3个平行）随机分配到长方形硬塑养殖箱（容积40×30×12cm3）中，利用充气式养殖系统保持海水溶氧量DO>6mg/L，pH6.5～7.5；水温24～28℃；盐度9.18～11.99 %。每天换1/3体积的水。采用自然光照，每日按照南美白对虾体重的5%投喂饲料，每日分为三次，早上9点一次，下午3点一次，晚上9点一次，每日饲料饲喂比为2：3：5。
随机挑选体质健康、个体大小均一的罗非鱼（鱼体重10±05g)，按照每组30尾（设3个平行）随机分配到长方形硬塑养殖箱（容积40×30×12cm3）中，水温：25～28 ℃，溶氧浓度＞5 mg /L，pH 7.8～8.2，氨氮浓度≤0.02 mg /L，亚硝酸盐浓度≤0.2 mg /L。采用自然光照，每日按照罗非鱼体重的5%，每日分为两次，早上8点一次，下午4点一次，每日饲料饲喂比为1：1。
以饲料中含不同剂量的T-2毒素（0、4.8、7.2、10.8、16.2和24.3 mg/kg·feed）、AFB1毒素（0、1.2、1.8、2.7、4.1和6.1 mg/kg·feed）、DON毒素（0、6.0、9.0、13.5、20.3和30.4 mg/kg·feed）和OTA毒素（0、1.78、2.67、4.01、6.01和9.01 mg/kg·feed）分别对对虾/罗非鱼进行递增剂量法染毒试验，每天进行染毒，4天为一周期，每期递增1.5倍，连续染毒20天后，得到染真菌毒素活体对虾/罗非鱼。
（2）饲喂外源乳酸菌
含嗜酸乳杆菌的对虾/罗非鱼饵料的制备：挑取嗜酸乳杆菌在MRS肉汤培养基中无氧培养26～30h（乳酸菌量达1.2×109～6×109 CFU/mL），按1：1的比例均匀喷洒至对虾/罗非鱼基础饵料中（配制成嗜酸乳杆菌在鱼/虾饲料，饲料中嗜酸乳杆菌的含量为1×108～3.2×108 CFU/g·feed），于37℃干燥。
将对虾/罗非鱼染毒20天结束后，开始投喂含嗜酸乳杆菌的对虾/罗非鱼饵料，投喂方式和比例同步骤（1），连续投喂10天，以继续投喂基础饲料为对照组。每两天取样一次，在无菌超净台解剖，得到对虾/罗非鱼肌肉和肠道，于-20 ℃保存。
采用LC-MS/MS方法检测对虾/罗非鱼肌肉中真菌毒素的残留量，结果发现：与对照组相比，后面10天投喂嗜酸乳杆菌的对虾/罗非鱼饵料组的对虾/罗非鱼肌肉中的真菌毒素残留量显著下降：对虾肌肉中T-2毒素残留量下降63.86±1.32%，AFB1下降53.04±0.97%，DON下降70.74±1.47%，OTA下降70.24±1.22%（图3）；罗非鱼肌肉中T-2毒素下降54.93±2.52%，AFB1下降42.18±1.97%，DON下降48.75±0.93%，OTA下降32.79±1.81%（图4），具体数据见表1和表2。
用MRS培养基对对虾/罗非鱼肠道微生物进行菌落计数，以实验组比上对照组得到微生物相对量比（即乳酸菌量比），发现乳酸菌量比与对虾/罗非鱼肌肉毒素消减速率成正比（图3和图4）。
Claims (5)
1.嗜酸乳杆菌在降解动物体内真菌毒素中的应用，其特征在于，所述真菌毒素为T-2毒素、黄曲霉毒素B1、呕吐毒素和赭曲霉毒素。
2.一种利用嗜酸乳杆菌降解动物体内真菌毒素的方法，其特征在于，是将嗜酸乳杆菌菌液与动物饲料混合后饲喂动物。
3.根据权利要求2所述的利用嗜酸乳杆菌降解动物体内真菌毒素的方法，其特征在于，所述嗜酸乳杆菌菌液的体积与动物饲料的质量比为1：1。
4.根据权利要求3所述的利用嗜酸乳杆菌降解动物体内真菌毒素的方法，其特征在于，所述嗜酸乳杆菌菌液中菌含量为1.2×109～6×109CFU/mL。
5.根据权利要求2至4任一项所述的利用嗜酸乳杆菌降解动物体内真菌毒素的方法，其特征在于，所述动物为罗非鱼和/或对虾。
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