Earticle

초록

영어

When researchers performed intestinal microbiota analysis using fecal samples, sample preparation processes are not standardized as of now. In particular, contrary to lab conditions, there are various factors hard to be controlled when sampling livestock feces on the farm. In this study, we wanted to know the influences of sample preparation conditions on fecal microbiota. We designed an experiment considering various sample preparation conditions (sample origin: rectum and ground; transporting condition: lysis buffer-treated, dry ice, ice, and room temperature; delayed times to extract DNA: 0, 1, 6, and 24 h) that can occur during microbiota analysis using cattle feces. First, we investigated the influences of sample origin, and microbial diversity (observed OTUs, p<0.001) was significantly higher when fecal samples were obtained from the ground than from the rectum and the principal coordinates analysis plot showed that samples were divided into two groups by origin. When we investigated the influences of transporting and storage conditions, observed OTUs (p<0.05) was significantly higher when samples were transported at room temperature than other conditions, and microbiota was affected by transporting and storage conditions. Finally, we investigated the effects of delayed time before DNA extraction from frozen fecal samples on microbial composition. Although this factor did not have a significant influence on microbial diversity, multidimensional scaling revealed its impact on microbial composition. Our findings will contribute to establishing an optimal procedure for microbiota analysis using farm-housing livestock feces.

목차

ABSTRACT
Ⅰ. INTRODUCTION
Ⅱ. MATERIALS AND METHODS
1. Sample preparation
2. DNA extraction and sequencing
3. Microbiota analysis
4. Statistical analyses
Ⅲ. RESULTS
1. Effects of sample origin on fecal microbiota
2. Effects of transporting method and storage condition on fecal microbiota
3. Effects of delayed time before DNA extraction from frozen samples on microbial composition
Ⅳ. DISCUSSION
Conflict of interest
ACKNOWLEDGEMENTS
Author’s Contributions
Ethics approval and consent to participate
Ⅴ. REFERENCES

저자

• 이상목 [ Sang-Mok Lee | 서울대학교 농생명공학부 대학원생 ]
• 한건구 [ Geongoo Han | 서울대학교 농생명공학부 대학원생 ]
• 김은배 [ Eun Bae Kim | 강원대학교 동물생명과학대학 동물응용과학과 교수 ]
• 강상기 [ Sang-Kee Kang | 서울대학교 그린바이오과학기술연구원 교수;서울대학교 국제농업기술대학원 교수 ] Corresponding Author

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