CAJ | 학술논문

为了比较奶牛瘤胃中不同粗饲料粘附细菌菌群结构,选择3头体况相近、健康、装有永久性瘤胃瘘管的荷斯坦奶牛,将苜蓿与全株玉米青贮混合粗饲料日粮(MF)和单一玉米秸秆日粮(CS)分别装入尼龙袋,在瘤胃中孵育24 h后取出,用磷酸缓冲液洗脱获得饲料固相粘附部分,剩余的饲料残渣是固相超紧密粘附部分,提取总 DNA,最终获得固相紧密粘附细菌和固相超紧密粘附细菌,利用 PCR-DGGE 分析细菌群落结构.结果表明：不同粘附程度的固相细菌的DGGE图谱条带数目和颜色深浅存在较大的差异,而粘附程度相同的不同粗饲料的 DGGE 图谱条带相似.2种饲料的瘤胃固相紧密粘附细菌的 Shannon-Wiener多样性指数,CS高于 MF(P<0.05),相比于固相紧密粘附细菌,固相超紧密粘附细菌的 Shannon-Wiener多样性指数更高(P<0.01).不同粘附程度的固相细菌差异条带近源种分别归属于Brevundimonas ,Prevotella,Pseudobutyrivibrio ,Clostridium.因此认为,粗饲料类型和粘附程度影响细菌多样性,其中固相超紧密粘附细菌的多样性较高.
위료비교내우류위중불동조사료점부세균균군결구,선택3두체황상근、건강、장유영구성류위루관적하사탄내우,장목숙여전주옥미청저혼합조사료일량(MF)화단일옥미갈간일량(CS)분별장입니룡대,재류위중부육24 h후취출,용린산완충액세탈획득사료고상점부부분,잉여적사료잔사시고상초긴밀점부부분,제취총 DNA,최종획득고상긴밀점부세균화고상초긴밀점부세균,이용 PCR-DGGE 분석세균군락결구.결과표명：불동점부정도적고상세균적DGGE도보조대수목화안색심천존재교대적차이,이점부정도상동적불동조사료적 DGGE 도보조대상사.2충사료적류위고상긴밀점부세균적 Shannon-Wiener다양성지수,CS고우 MF(P<0.05),상비우고상긴밀점부세균,고상초긴밀점부세균적 Shannon-Wiener다양성지수경고(P<0.01).불동점부정도적고상세균차이조대근원충분별귀속우Brevundimonas ,Prevotella,Pseudobutyrivibrio ,Clostridium.인차인위,조사료류형화점부정도영향세균다양성,기중고상초긴밀점부세균적다양성교고.
In order to compare rumen bacterial communities adherent to different types of forages,three healthy Holstein dairy cows with permanent rumen cannula and similar weight were selected.Two types of feeds,mixed forage (alfalfa hay and silage corn,MF)and corn straw(CS)were put into nylon bags and in-cubated in the rumen for 24 h.The solid adherent fraction of feeds was prepared by PBS elution,and the residue was solid super-adherent fraction.After DNA extraction from different fractions,DGGE was intro-duced to compare the bacterial diversity.The results showed that the number and intensity of bands from DGGE profile in different fractions were different,that in different forages from the same fraction were similar.The bacterial diversity index of Shannon-Wiener in adherent fraction from CS was higher compared with MF (P<0.05),and super-adherent fraction showed higher bacterial diversity compared with adherent fraction.The different bacteria in different fractions belonged to Brevundimonas ,Prevotella ,P seudobu-tyrivibrio and Clostridium.It was concluded that different types of forages and adherent fractions affected bacterial diversity,and the bacteria in super-adherent fraction from MF exhibited higher diversity.