The objective of this study was to investigate the effect of different feed inoculation method on rumen fermentation in an in vitro. Three experimental treatments were used: control (CON, direct dispersion of feed (2 g) in rumen fluid), combinations of direct dispersion (1 g) and nylon bag (DNB, pore size: 50 μm, 1 g), and nylon bag (NB, 2 g). An in vitro fermentation experiment was carried out using strained rumen fluid for 48 h incubation time and timothy was used as a substrate. At the end of the incubation, in vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), pH, volatile fatty acids (VFA), ammonia nitrogen (NH3-N), and microbial community were evaluated and gas production was estimated at 3, 6, 12, 24, 48 h incubation periods. Gas production was higher in CON than DNB and NB at 6 and 12 h incubation time (p<0.01). There were no differences in final gas production, pH, NH3-N concentration, total VFA production, and VFA profiles among treatments. The IVDMD was lowest in CON (p<0.01) but the IVNDFD was not differed by feed distribution methods. There were no significant differences in general bacteria and fungi. Protozoa count was highest in NB treatment among treatments (p<0.01). The abundance of cellulolytic bacteria, Ruminococcus flavefaciens and Fibrobacter succinogenes, was highest in the CON among treatments (p<0.01).

ABSTRACT
Ⅰ. 서론
Ⅱ. 재료 및 방법
    1. 공시재료 및 공시동물
    2. 영양소 성분분석
    3. In vitro 발효 평가
    4. genomic DNA 추출
    5. 재조합 plasmid의 제작
    6. Real-time PCR
    7. Gas parameter
    8. 통계 분석
Ⅲ. 결과 및 고찰
Ⅳ. 요약
Ⅴ. 참고문헌

• 유대겸(부산대학교 생명자원과학대학 동물생명자원과학과) | Dae-Kyum Yoo (Life and Industry Convergence Research Institute, Department of Animal Science, Pusan National University)
• 문준범(부산대학교 생명자원과학대학 동물생명자원과학과) | Joon-Beom Moon (Life and Industry Convergence Research Institute, Department of Animal Science, Pusan National University)
• 김한빈(부산대학교 생명자원과학대학 동물생명자원과학과) | Han-Been Kim (Life and Industry Convergence Research Institute, Department of Animal Science, Pusan National University)
• 양성재(부산대학교 생명자원과학대학 동물생명자원과학과) | Sung-Jae Yang (Life and Industry Convergence Research Institute, Department of Animal Science, Pusan National University)
• 박중국(농협사료) | Joong-Kook Park (Nonghyupfeed Co., Ltd)
• 이세영(천안연암대학교 축산계열) | Se-Young Lee (Department of Animal Science, Yonam College)
• 서자겸(부산대학교 생명자원과학대학 동물생명자원과학과) | Ja-Kyeom Seo (Life and Industry Convergence Research Institute, Department of Animal Science, Pusan National University) Corresponding Author