不同缓释速度瘤胃调控剂对瘤胃亚急性酸中毒体外发酵参数的影响
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摘要
本论文通过三个试验探讨了制备不同释放速度瘤胃调控剂对瘤胃亚急性酸中毒体外发酵参数的影响。首先通过采用玉米和鲜酒糟在体外发酵构建瘤胃亚急性酸中毒(Subacute Rumen Acidosis, SARA)模型,然后利用碳酸氢钠、氧化镁、烟酸、硫酸锌、硫酸钴以及高分子骨架材料羟丙基甲基纤维素(HPMC-K100M),辅料如淀粉、无水乙醇、硬脂酸镁等制备了6h、8h和10h完全释放的瘤胃调控剂,最后考察了不同释放速度的瘤胃调控剂对体外业急性酸中毒模型中瘤胃发酵参数的影响。本研究分为三个部分。
试验一体外瘤胃亚急性酸中毒模型的建立
本试验选取3头体重350kg左右、安装有永久性瘤胃瘘管的宣汉黄牛,于晨饲2h后采集瘤胃内容物,滤液作为体外发酵液的接种物,通过添加玉米与鲜酒糟的方式来诱导体外发生SARA。试验采用对照试验设计,分为两个处理组：A组(玉米：鲜酒糟=20：80)、B组(玉米：鲜酒糟=50：50)。结果表明：①随着玉米／鲜酒糟比例的提高,各处理组瘤胃液内pH显著降低,瘤胃液pH值低于5.5的持续时间也随之延长。②B组(玉米：鲜酒糟=50：50)在6h和18h时pH值分别达到了5.6和5.3,pH值低于5.6的持续时间超过了10h以上,成功构建了体外瘤胃亚急性酸中毒模型。试验二不同释放速度瘤胃调控剂的制备和释放度的测定
本试验选取碳酸氢钠、氧化镁,烟酸、硫酸钴、硫酸锌,与羟丙基甲基纤维素,无水乙醇等混匀制备瘤胃调控剂,并根据中国药典二部附录XD释放度测定第一法“转篮法”测定缓释片剂体外释放度。结果表明：①由羟丙基甲基纤维素(HPMC-K100M)包被制备的瘤胃调控剂体外释放度在8h时达到了82.06%,优于HPMC-K4M和HPMC-K15M,更适宜于做本试验中的包被材料。②依据中国药典规定,分别添加2%、2.5%和3%的HPMC-K100M制备的瘤胃调控剂在6h、8h、10h内释放度分别为80.75%、80.25%和82.50%,能够制备6h、8h、10h完全释放的瘤胃调控剂。
实验三不同释放速度瘤胃调控剂对亚急性酸中毒瘤胃发酵参数的影响
本试验借助于构建成功的SARA体外模型,设置4个处理组：对照组(不采用包被材料的瘤胃调控剂),6h完全释放组,8h完全释放组和10h完全释放组。在成功构建SARA模型之后添加不同释放速度的瘤胃调控剂,以此时为0时间点、并在随后的2、4、6、8、10、12 h取样测定瘤胃液内各发酵指标及牛链球菌的相对数量。结果表明,各组都显著提高了瘤胃液pH值(P<0.01),乙酸、丙酸、丁酸、乳酸和总挥发性脂肪酸(Total Volatile Fatty Acids, TVFA)含量显著增加(P<0.05)。8h释放组乳酸含量在6h达到1.23 mmol/L之后逐渐增加,在12h时达到最大值1.34 mmol/L,显著高于其他各组(P<0.01)。牛链球菌占全部细菌的相对含量变化与乳酸含量结果变化情况相似。8h完全释放组牛链球菌相对含量逐渐增加,在12h时达到了最大值1.06%,显著高于其他各组(P<0.01)。8h完全释放组有减缓瘤胃液内内毒素和组胺生成的趋势,而6h完全释放组和对照由于较快的释放速度改变了瘤胃微生物区系稳态,内毒素含量在10h分别达到了18,197 EU/ml和33,113 EU/ml;组胺含量在10h分别达到了42.13ng/ml和48.10ng/ml.
上述试验结果表明：8h完全释放组有效缓解了瘤胃亚急性酸中毒过程中较低的pH值,减缓了瘤胃内挥发性脂肪酸(乙酸、丙酸、丁酸)及乳酸的生成,同时瘤胃液组胺、内毒素的含量也显著下降,有效的改变了瘤胃的发酵特性。
The aim of this study was to evaluate the effects of different releasing rates of rumen modulators on in vitro fermentation characteristics and S. bovis number by the means of induced SARA model. The paper had three parts:
Part1. The development of SARA model
We used three Xuanhan steers fitted with permanent ruminal cannulas and SARA was induced by adding corn and fresh distilled grain(1:1). And this experiment was divided into 2 groups:group A (Corn/Fresh distilled grain=80:20) and group B(Corn/Fresh distilled grain=50:50). It was found that rumen pH decreased significantly with the proportion of corn/fresh distilled grain increased as well as the time below 5.5. The results also showed that rumen pH needed much longer time to reach 5.5 in group A, while rumen pH of group B reached 5.5 in 7h and duration below 5.5 was more than 8h. Therefore, when added corn and fresh distilled grain(1:1), SARA was induced successfully.
Part2 Preparation of different release rate of rumen modulators and the evaluation of accumulative releasing rates.
Sodium bicarbonate, magnesium oxide, nicotinic acid(NA), cobalt sulphate, zinc sulfate and starch, hydroxypropyl methyl cellulose(HPMC) and absolute ethyl alcohol were mixed to prepared for the modulators and evaluated their accumulative releasing rates. Finally,6h release group,8h release group and 10h release group were successfully prepared. The results showed that HPMC could regulate the releasing rates of kalinous materials and the accumulative releasing rates met the requirements of this design.
Part3 The effects of different releasing rates of rumen modulators on in vitro fermentation characteristics and S. bovis number
试验一体外瘤胃亚急性酸中毒模型的建立
本试验选取3头体重350kg左右、安装有永久性瘤胃瘘管的宣汉黄牛,于晨饲2h后采集瘤胃内容物,滤液作为体外发酵液的接种物,通过添加玉米与鲜酒糟的方式来诱导体外发生SARA。试验采用对照试验设计,分为两个处理组：A组(玉米：鲜酒糟=20：80)、B组(玉米：鲜酒糟=50：50)。结果表明：①随着玉米／鲜酒糟比例的提高,各处理组瘤胃液内pH显著降低,瘤胃液pH值低于5.5的持续时间也随之延长。②B组(玉米：鲜酒糟=50：50)在6h和18h时pH值分别达到了5.6和5.3,pH值低于5.6的持续时间超过了10h以上,成功构建了体外瘤胃亚急性酸中毒模型。试验二不同释放速度瘤胃调控剂的制备和释放度的测定
本试验选取碳酸氢钠、氧化镁,烟酸、硫酸钴、硫酸锌,与羟丙基甲基纤维素,无水乙醇等混匀制备瘤胃调控剂,并根据中国药典二部附录XD释放度测定第一法“转篮法”测定缓释片剂体外释放度。结果表明：①由羟丙基甲基纤维素(HPMC-K100M)包被制备的瘤胃调控剂体外释放度在8h时达到了82.06%,优于HPMC-K4M和HPMC-K15M,更适宜于做本试验中的包被材料。②依据中国药典规定,分别添加2%、2.5%和3%的HPMC-K100M制备的瘤胃调控剂在6h、8h、10h内释放度分别为80.75%、80.25%和82.50%,能够制备6h、8h、10h完全释放的瘤胃调控剂。
实验三不同释放速度瘤胃调控剂对亚急性酸中毒瘤胃发酵参数的影响
本试验借助于构建成功的SARA体外模型,设置4个处理组：对照组(不采用包被材料的瘤胃调控剂),6h完全释放组,8h完全释放组和10h完全释放组。在成功构建SARA模型之后添加不同释放速度的瘤胃调控剂,以此时为0时间点、并在随后的2、4、6、8、10、12 h取样测定瘤胃液内各发酵指标及牛链球菌的相对数量。结果表明,各组都显著提高了瘤胃液pH值(P<0.01),乙酸、丙酸、丁酸、乳酸和总挥发性脂肪酸(Total Volatile Fatty Acids, TVFA)含量显著增加(P<0.05)。8h释放组乳酸含量在6h达到1.23 mmol/L之后逐渐增加,在12h时达到最大值1.34 mmol/L,显著高于其他各组(P<0.01)。牛链球菌占全部细菌的相对含量变化与乳酸含量结果变化情况相似。8h完全释放组牛链球菌相对含量逐渐增加,在12h时达到了最大值1.06%,显著高于其他各组(P<0.01)。8h完全释放组有减缓瘤胃液内内毒素和组胺生成的趋势,而6h完全释放组和对照由于较快的释放速度改变了瘤胃微生物区系稳态,内毒素含量在10h分别达到了18,197 EU/ml和33,113 EU/ml;组胺含量在10h分别达到了42.13ng/ml和48.10ng/ml.
上述试验结果表明：8h完全释放组有效缓解了瘤胃亚急性酸中毒过程中较低的pH值,减缓了瘤胃内挥发性脂肪酸(乙酸、丙酸、丁酸)及乳酸的生成,同时瘤胃液组胺、内毒素的含量也显著下降,有效的改变了瘤胃的发酵特性。
The aim of this study was to evaluate the effects of different releasing rates of rumen modulators on in vitro fermentation characteristics and S. bovis number by the means of induced SARA model. The paper had three parts:
Part1. The development of SARA model
We used three Xuanhan steers fitted with permanent ruminal cannulas and SARA was induced by adding corn and fresh distilled grain(1:1). And this experiment was divided into 2 groups:group A (Corn/Fresh distilled grain=80:20) and group B(Corn/Fresh distilled grain=50:50). It was found that rumen pH decreased significantly with the proportion of corn/fresh distilled grain increased as well as the time below 5.5. The results also showed that rumen pH needed much longer time to reach 5.5 in group A, while rumen pH of group B reached 5.5 in 7h and duration below 5.5 was more than 8h. Therefore, when added corn and fresh distilled grain(1:1), SARA was induced successfully.
Part2 Preparation of different release rate of rumen modulators and the evaluation of accumulative releasing rates.
Sodium bicarbonate, magnesium oxide, nicotinic acid(NA), cobalt sulphate, zinc sulfate and starch, hydroxypropyl methyl cellulose(HPMC) and absolute ethyl alcohol were mixed to prepared for the modulators and evaluated their accumulative releasing rates. Finally,6h release group,8h release group and 10h release group were successfully prepared. The results showed that HPMC could regulate the releasing rates of kalinous materials and the accumulative releasing rates met the requirements of this design.
Part3 The effects of different releasing rates of rumen modulators on in vitro fermentation characteristics and S. bovis number
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