CAJ | 학술논문

本试验旨在应用高通量测序技术比较山羊瘤胃和粪便微生物的结构与组成。选取6只10月龄波尔山羊,饲喂精粗比为3∶7的饲粮14 d后采集瘤胃液(R组)和粪便样品(F组)(每组6个重复,每个样品为1个重复)。提取总DNA后用古菌/细菌16S rRNA通用引物扩增V4~V5区,并用 Illumina MiSeq 平台测序。结果表明：1)共获得有效序列227729条,聚类后得13601个运算分类单位(OTU)。2)所得OTU经物种注释99.337%被归类为细菌界,F组相对丰度最高优势菌门为厚壁菌门(65.400%),R组相对丰度最高优势菌门为拟杆菌门(60.188%)。3)从所有样品中共检测到129个科,F组中相对丰度最高为瘤胃球菌科(37.705%),极显著高于R组(P<0.01),而R组中,相对丰度最高的为普雷沃氏菌科(29.959%),极显著高于F组(P<0.01)。4)在粪便样品和瘤胃液样品种共检测到258个属,F组相对丰度最高的属为瘤胃球菌科未分类的属(26.914%),R组相对丰度最高为普雷沃菌属(28.621%)。5)所有12个样品间共发现了14个共享属,其中相对丰度最高的为厚壁菌门下的梭菌属4( Clostridium_Ⅳ,1.748%)。本试验结果表明瘤胃和粪便中微生物组成存在着较大差异,瘤胃中还有许多未被分类鉴定且相对丰度较高的微生物,需要进一步研究。
본시험지재응용고통량측서기술비교산양류위화분편미생물적결구여조성。선취6지10월령파이산양,사위정조비위3∶7적사량14 d후채집류위액(R조)화분편양품(F조)(매조6개중복,매개양품위1개중복)。제취총DNA후용고균/세균16S rRNA통용인물확증V4~V5구,병용 Illumina MiSeq 평태측서。결과표명：1)공획득유효서렬227729조,취류후득13601개운산분류단위(OTU)。2)소득OTU경물충주석99.337%피귀류위세균계,F조상대봉도최고우세균문위후벽균문(65.400%),R조상대봉도최고우세균문위의간균문(60.188%)。3)종소유양품중공검측도129개과,F조중상대봉도최고위류위구균과(37.705%),겁현저고우R조(P<0.01),이R조중,상대봉도최고적위보뢰옥씨균과(29.959%),겁현저고우F조(P<0.01)。4)재분편양품화류위액양품충공검측도258개속,F조상대봉도최고적속위류위구균과미분류적속(26.914%),R조상대봉도최고위보뢰옥균속(28.621%)。5)소유12개양품간공발현료14개공향속,기중상대봉도최고적위후벽균문하적사균속4( Clostridium_Ⅳ,1.748%)。본시험결과표명류위화분편중미생물조성존재착교대차이,류위중환유허다미피분류감정차상대봉도교고적미생물,수요진일보연구。
This experiment was conducted to explore structure and composition of ruminal and fecal microbiota of goat. Six ten-month-old Boer goats were fed a diet consisting of 30% concentrate and 70% roughage. After 14-day experiment, the rumen fluid ( R group) and feces ( F group) were sampled [ each group had 3 repli-cates ( samples) ] . Total DNA of rumen content and feces samples were extracted, and universal prokaryote primers were used to target the V4 to V5 hypervariable region of 16S rRNA, finally the products were se-quenced on MiSeq Illumina sequencing platform. The results showed as follows: 1 ) a total of 227 729 se-quences across all rumen content and fecal samples were generated, and the total number of operational taxo-nomic units ( OTUs) detected by the cluster analysis reached 13 601. 2) The 99.337% of all OTUs belonged to bacteria. The most abundant phyla in F and R groups was Firmicutes ( 65. 400%) and Bacteroidetes ( 60.188%) , respectively. 3) One hundred and twenty nine families were detected in all samples. The most a-bundant families found in F group was Ruminococcaceae (37.705%), which was extremely significantly high-er than that in R group ( P<0. 01 ); the most abundant families found in R group was Prevotellaceae (29.959%), which was extremely significantly higher than that in F group (P<0.01). 4) Two hundred and twenty nine genera were detected in all samples. The most abundant genus found in F and R groups was unclas-sified Ruminococcaceae (26.914%) and Prevotella (28.621%), respectively. 5) The analysis revealed 14 genera shared by all 12 samples. Among them, the most abundant genus was Clostridium_Ⅳ(1.748%). It is concluded that there is a significant difference between ruminal and fecal microbiota, and there are many un-classified microbiomes that have high relative abundance in the rumen, which need further investigations.