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PCR-DGGE analysis of silage microorganisms and its application to food quality control

青贮饲料微生物PCR-DGGE分析及其在食品质量控制中的应用

基本信息

批准号：
17580236
负责人：
NAOKI Nishino
金额：
$ 2.3万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17580236/
关键词：
Feed Microbiology PCR-DGGE Silage

项目摘要

It has become a common practice in Japan that high-moisture by-products are stored with dry feeds as an total mixed ration silage. This may minimize the risk of effluent production and can omit the time for mixing prior to feeding. Unpalatable by-products could be incorporated into a TMR silage, if their odors and flavors are altered through the fermentation in a silo. An interesting property of the TMR silage is high stability on exposure to air. Heating would not occur even in summer, whereas non-ensiled TMR mixture may deteriorate quite rapidly. Conventional criteria do not account well for the stability, because the TMR silage can remain unheated even with a large number of yeasts and a substantial level of substrates (sugars and lactic acid) for the growth of aerobic microorganisms. We have investigated the factors involved in resistance to deterioration by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), and some interesting results are obtained as fo … More llows.1. When wet brewers grains and soybean-curd refuse were stored alone or in combination with other feeds as a TMR silage, deterioration did not occur only in TMR silages. Without anaerobic fermentation, TMR mixture also spoiled quite easily.2. Lactobacillus buchneri, a hetero-fermentative lactic acid bacteria which is capable of enhancing aerobic stability, was not found in the silage material, but appeared from the early ensiling period in TMR silage containing wet brewers grains. Although aerobic stability was similarly high in TMR silage containing soybean-curd refuse, L. buchneri was not detected regardless of ensiling period.3. Weisella sp. and Lactobacillus brevis was found in both the stable TMR silages. Unstable by-products silage stored alone did not contain these two lactic acid bacteria.4. We could isolate those L. buchneri, L. brevis and Weisella confusa from the following experiment, hence now are planning to inoculation experiment to determine if these bacterial mixture can actually improve the aerobic stability of silage. Less

在日本，高水分副产品与干饲料一起作为全混合日粮青贮饲料储存已成为一种常见做法。这可以最大限度地降低产生废水的风险，并且可以省去进料前的混合时间。如果在筒仓中发酵改变了难闻的副产品的气味和味道，则可以将其纳入 TMR 青贮饲料中。 TMR 青贮饲料的一个有趣特性是暴露在空气中的高稳定性。即使在夏天也不会加热，而未青贮的 TMR 混合物可能会很快变质。传统标准并不能很好地解释稳定性，因为即使存在大量酵母和大量用于需氧微生物生长的底物（糖和乳酸），TMR 青贮饲料也可以保持不加热状态。我们通过聚合酶链反应-变性梯度凝胶电泳（PCR-DGGE）研究了抗变质的因素，并获得了一些有趣的结果，如下所示。 1．当湿啤酒谷物和豆腐渣单独储存或与其他饲料混合作为TMR青贮饲料储存时，变质不仅仅发生在TMR青贮饲料中。如果不进行厌氧发酵，TMR混合物也很容易变质。2．布氏乳杆菌是一种能够增强有氧稳定性的异质发酵乳酸菌，在青贮饲料材料中未发现，但在含有湿啤酒谷物的TMR青贮饲料中在青贮早期就出现了。尽管含有豆腐渣的TMR青贮饲料的有氧稳定性同样较高，但无论青贮期如何，均未检测到L. buchneri。3．威塞拉属在两种稳定的 TMR 青贮饲料中都发现了短乳杆菌。单独储存的不稳定副产品青贮饲料中不含这两种乳酸菌。4．我们可以从下面的实验中分离出L. buchneri、L. brevis和Weisella conusa，因此现在计划进行接种实验来确定这些细菌混合物是否确实可以提高青贮饲料的有氧稳定性。较少的