应用饲料油脂防控牛感染大肠杆菌0157：H7的研究
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摘要
本论文研究了亚麻籽和中链脂肪酸用于防控牛感染大肠杆菌O157:H7的可行性。试验分为两部分,第一部分通过向牛日粮中添加亚麻籽并取瘤胃液和真胃液与大肠杆菌O157:H7进行体外培养以验证长链脂肪酸是否在反刍动物瘤胃和真胃中对大肠杆菌O157:H7有抑制作用,同时本试验还通过检测不同日粮添加亚麻籽对牛胃肠道内容物中脂肪酸的组成及牛肠道内容物中粘膜抗体免疫球蛋白A浓度的影响以确定长链脂肪酸对牛肠道免疫力的影响,本论文还研究了日粮中添加亚麻籽对肉牛生产性能的影响从而为脂肪酸的应用奠定基础。第二部分为了更好的使用脂肪酸抵抗大肠杆菌O157:H7感染,本论文通过体外培养研究了不同碳链长度脂肪酸在不同pH条件下对大肠杆菌O157:H7的抑制活性,及中链脂肪酸与亚麻籽在瘤胃液和真胃液中的协同抑菌效果,从而为脂肪酸的合理搭配提供理论基础。试验结果如下:
1不同日粮添加亚麻籽对牛瘤胃液和真胃液抑菌活性的影响不同。饲喂青干草+大麦组牛瘤胃液中添加亚麻籽显著抑制了大肠杆菌O157:H7的繁殖和真胃液中大肠杆菌O157:H7的存活,而青贮+大麦组牛添加亚麻籽后瘤胃液抑菌活性并未增强,但其真胃液抑菌活性显著提高。不同日粮添加亚麻籽对牛胃肠道内容物脂肪酸组成和IgA含量的影响不同,青贮+大麦日粮添加亚麻籽提高了牛十二指肠、回肠和结肠粘膜抗体IgA含量。添加亚麻籽提高了饲料转化效率,并增加了牛皮下脂肪中亚麻酸含量。
2癸酸、月桂酸和亚油酸在pH2.5培养条件下抑菌活性最强,三种18碳不饱和脂肪酸在pH2.5培养条件下的抑大肠杆菌O157:H7活性为:亚油酸>油酸>亚麻酸,脂肪酸的抑菌活性随pH升高而降低。癸酸在青干草+大麦+亚麻籽组牛瘤胃液和真胃液中表现出与亚麻籽的协同抑菌效应。月桂酸在青干草+大麦+亚麻籽组牛瘤胃液中未表现出与亚麻籽的协同抑菌效应,但在真胃液中表现出与亚麻籽的协同抑菌效应。癸酸和月桂酸在饲喂青贮+大麦组牛的瘤胃液和真胃液中的抑菌活性强于其他各组,但未与亚麻籽表现出有协同抑菌效应。各组日粮牛真胃液调节到pH3.5后,中链脂肪酸的抑菌活性显著升高,癸酸在3h内将大肠杆菌降低到检测范围之下,月桂酸的抑菌活性也显著增强。
本试验结果表明,添加亚麻籽能够在一定程度上抑制瘤胃和真胃液中大肠杆菌O157:H7的数量,但如果同时加入中链脂肪酸则抑菌效果更佳。饲料添加亚麻籽同时能够提高牛肠道免疫性能和生产性能并改善牛肉的脂肪酸组成。本试验将脂肪酸的抑菌功能和提高生产性能的功能相结合,为开发绿色、无污染的生物防治手段提供了一条可行路线。
The aim of current research was to study the feasibility of using feed oil to eliminate the infection of Escherichia coli O157:H7 in cattle. The whole experiment was carried out by two trails. In first trail, the ruminal and abomasal content were sampled from cattle fed different diet with or without flaxseed and were used to do the in vitro culture with Escherichia coli O157:H7 to see the influence of feed oil on the pathogen. At the same time we analyzed the composition of the fatty acids and the concentration of the IgA in bovine gastro-intestinal content to see the influence of feed oil on the bovine intestinal immunity. In the second trail, we studied the antibacterial effect of different single fatty acid on E.coli O157:H7 in LB medium. We also studied the synergism antibacterial activity of medium chain fatty acid and long chain fatty acid in ruminal and abomasal fluid.
Our data indicated that addition of flaxseed to different diet has different influence on the antibacterial activity of bovine ruminal and abomasal fluid. Addition of flaxseed to the grass hay and barley diet increase the bactericidal activity of bovine ruminal and abomasal fluid, however addition of flaxseed to barley silage and barley diet only has influence on antibacterial activity of bovine abomasal fluid not on ruminal fluid. Addition of flaxseed changed the fatty acid composition of bovine gastro-intestinal content and increased the concentration of IgA which indicated the increased intestinal immunity of cattle.
In our experiment, capric acid, lauric acid and linoleic acid show the strongest antibacterial activity on E. coli O157:H7 when incubated in pH 2.5 LB medium. The order of anti-E.coli O157:H7 activity of C18 fatty acid at pH 2.5 was as follow: linoleic acid > oleic acid > linolenic acid. The bactericidal activity of fatty acid was decreased with the increase of pH. Capric acid and flaxseed show synergism anti-E. coli O157:H7 activity when added to ruminal and abomasal fluid from cattle fed hay and barley. However, Lauric acid and flaxseed only show synergism anti-E. coli O157:H7 activity when added to abomasal fluid from cattle fed same diet. Both capric and lauric acid did not show synergism anti-E. coli O157:H7 activity when added to ruminal and abomasal fluid from cattle fed barley silage and barley. The antibacterial activity of capric and lauric acid was increased when the bovine abomasal fluid pH decreased to 3.5.
In general, addition of flaxseed in bovine diet could partially increase the antibacterial activity of bovine ruminal and abomasal fluid, and this activity may be increased if added medium chain fatty acids at the same time. Addition of flaxseed could increase the immunity of bovine intestinal by increase the secretion of IgA. The immunity increasing function and the antibacterial activity of fatty acid was combinated in our study to give a new suggestion to eliminate the infection of cattle by E.coli O157:H7 without antibiotic resistance.
1不同日粮添加亚麻籽对牛瘤胃液和真胃液抑菌活性的影响不同。饲喂青干草+大麦组牛瘤胃液中添加亚麻籽显著抑制了大肠杆菌O157:H7的繁殖和真胃液中大肠杆菌O157:H7的存活,而青贮+大麦组牛添加亚麻籽后瘤胃液抑菌活性并未增强,但其真胃液抑菌活性显著提高。不同日粮添加亚麻籽对牛胃肠道内容物脂肪酸组成和IgA含量的影响不同,青贮+大麦日粮添加亚麻籽提高了牛十二指肠、回肠和结肠粘膜抗体IgA含量。添加亚麻籽提高了饲料转化效率,并增加了牛皮下脂肪中亚麻酸含量。
2癸酸、月桂酸和亚油酸在pH2.5培养条件下抑菌活性最强,三种18碳不饱和脂肪酸在pH2.5培养条件下的抑大肠杆菌O157:H7活性为:亚油酸>油酸>亚麻酸,脂肪酸的抑菌活性随pH升高而降低。癸酸在青干草+大麦+亚麻籽组牛瘤胃液和真胃液中表现出与亚麻籽的协同抑菌效应。月桂酸在青干草+大麦+亚麻籽组牛瘤胃液中未表现出与亚麻籽的协同抑菌效应,但在真胃液中表现出与亚麻籽的协同抑菌效应。癸酸和月桂酸在饲喂青贮+大麦组牛的瘤胃液和真胃液中的抑菌活性强于其他各组,但未与亚麻籽表现出有协同抑菌效应。各组日粮牛真胃液调节到pH3.5后,中链脂肪酸的抑菌活性显著升高,癸酸在3h内将大肠杆菌降低到检测范围之下,月桂酸的抑菌活性也显著增强。
本试验结果表明,添加亚麻籽能够在一定程度上抑制瘤胃和真胃液中大肠杆菌O157:H7的数量,但如果同时加入中链脂肪酸则抑菌效果更佳。饲料添加亚麻籽同时能够提高牛肠道免疫性能和生产性能并改善牛肉的脂肪酸组成。本试验将脂肪酸的抑菌功能和提高生产性能的功能相结合,为开发绿色、无污染的生物防治手段提供了一条可行路线。
The aim of current research was to study the feasibility of using feed oil to eliminate the infection of Escherichia coli O157:H7 in cattle. The whole experiment was carried out by two trails. In first trail, the ruminal and abomasal content were sampled from cattle fed different diet with or without flaxseed and were used to do the in vitro culture with Escherichia coli O157:H7 to see the influence of feed oil on the pathogen. At the same time we analyzed the composition of the fatty acids and the concentration of the IgA in bovine gastro-intestinal content to see the influence of feed oil on the bovine intestinal immunity. In the second trail, we studied the antibacterial effect of different single fatty acid on E.coli O157:H7 in LB medium. We also studied the synergism antibacterial activity of medium chain fatty acid and long chain fatty acid in ruminal and abomasal fluid.
Our data indicated that addition of flaxseed to different diet has different influence on the antibacterial activity of bovine ruminal and abomasal fluid. Addition of flaxseed to the grass hay and barley diet increase the bactericidal activity of bovine ruminal and abomasal fluid, however addition of flaxseed to barley silage and barley diet only has influence on antibacterial activity of bovine abomasal fluid not on ruminal fluid. Addition of flaxseed changed the fatty acid composition of bovine gastro-intestinal content and increased the concentration of IgA which indicated the increased intestinal immunity of cattle.
In our experiment, capric acid, lauric acid and linoleic acid show the strongest antibacterial activity on E. coli O157:H7 when incubated in pH 2.5 LB medium. The order of anti-E.coli O157:H7 activity of C18 fatty acid at pH 2.5 was as follow: linoleic acid > oleic acid > linolenic acid. The bactericidal activity of fatty acid was decreased with the increase of pH. Capric acid and flaxseed show synergism anti-E. coli O157:H7 activity when added to ruminal and abomasal fluid from cattle fed hay and barley. However, Lauric acid and flaxseed only show synergism anti-E. coli O157:H7 activity when added to abomasal fluid from cattle fed same diet. Both capric and lauric acid did not show synergism anti-E. coli O157:H7 activity when added to ruminal and abomasal fluid from cattle fed barley silage and barley. The antibacterial activity of capric and lauric acid was increased when the bovine abomasal fluid pH decreased to 3.5.
In general, addition of flaxseed in bovine diet could partially increase the antibacterial activity of bovine ruminal and abomasal fluid, and this activity may be increased if added medium chain fatty acids at the same time. Addition of flaxseed could increase the immunity of bovine intestinal by increase the secretion of IgA. The immunity increasing function and the antibacterial activity of fatty acid was combinated in our study to give a new suggestion to eliminate the infection of cattle by E.coli O157:H7 without antibiotic resistance.
引文
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