초록 ▼

○ 연구결과
다양한 근원으로부터 분리된 균중 내산성과 내담즙성 및 박테리오신 생산능력이 Lactobacillus acidophilus 107A를 선발 하였고, 대량생산시 균주의 최적의 성장을 위한 배지의 조건을 결정하였다. 분체복합화 공정 시 최적의 피복을 할수 있는 균주의 최적의 입도를 실험적으로 찾아내었고, 특히 반응표면방법에 입각하여 피복제와 생균제의 최적의 혼합비율은 1:9로 나타났다. 분체복합화된 생균제를 이용한 이유자돈과 육성돈의 사양실험에서는 분체복합 생균제 또는 생균제 첨가구가 항생제를 급여한 첨가부와 비슷한 일

○ 연구결과
다양한 근원으로부터 분리된 균중 내산성과 내담즙성 및 박테리오신 생산능력이 Lactobacillus acidophilus 107A를 선발 하였고, 대량생산시 균주의 최적의 성장을 위한 배지의 조건을 결정하였다. 분체복합화 공정 시 최적의 피복을 할수 있는 균주의 최적의 입도를 실험적으로 찾아내었고, 특히 반응표면방법에 입각하여 피복제와 생균제의 최적의 혼합비율은 1:9로 나타났다. 분체복합화된 생균제를 이용한 이유자돈과 육성돈의 사양실험에서는 분체복합 생균제 또는 생균제 첨가구가 항생제를 급여한 첨가부와 비슷한 일당증체량을 나타내어, 성장개선효과가 입증되었다. 또한, 생균제 급여후 도체의 육질분석을 통하여 항생제와 비슷하거나 더 좋을 결과를 얻을수 있었다.

Abstract ▼

In order to develop a probiotic product for promoting growth performance of pigs, many strains of microbials were isolated from various sources. Many laboratory tests, such as acid tolerance, bile tolerance, and production of bacterocin for anti-microbial resistance were conducted to select reliable

In order to develop a probiotic product for promoting growth performance of pigs, many strains of microbials were isolated from various sources. Many laboratory tests, such as acid tolerance, bile tolerance, and production of bacterocin for anti-microbial resistance were conducted to select reliable strains. As a result, seventeen strains of microbials were selected to be coated with prebiotics.
To optimize growth of microbials, Response Surface Methodology was used to develop a optimum medium. Skim milk, glucose, and yeast extract were considered as a major ingredient for microbials and the composition of medium was determined for selected strains. Response Surface Methodological figures of L. acidophilus 107A and L. acidophilus 393, which showed a relatively good property during selection, were visualized. And, to optimize for mass production, the viabilities of strains were determined when microbials were concentrated, frozen and stored. The L. casei 910, L. casei ssp., L. rahmnosus 7469, L. acidophilus 107A, and L. acidophilus 393 showed a reliable viability. The change of pH and color of broth and agar plate, which contained different prebiotics, and the viable cell counts were investigated to confirm the effects of prebiotics. The change of pH showed more accurate results than the change of color or the viable cell counts. Suitable strains were selected under addition of prebiotics. The Response Surface Methodology was used to determine the optimal concentration of prebiotics to maximize the synergy effects of mixture of probiotics and prebiotics. It was revealed that the statical methods for determine the medium recipe for microbial had reliable results.
The stability of microbial was investigated during stored at different temperatures, 4℃, 25℃, and 37℃. The stability of microbial was better at low temperature (4℃ than high temperature(25℃and 37℃). To make a optimize stock solution for long storage in frozen, the materials for stock and the ratio among materials were determined. Physio-chemical methods were used to determine the strains of isolated microbials. To confirm strains base on genomic level, the intergenic spacer region between 16S rRNA and 23S rRNA were sequenced. The
strains were confirmed by comparing the sequence to National Center for Biotechnology Information and Gen Bank. The anti-microbial Spectrum and viable cell counts were investigated for other effects. The adherence of selected strains against intestinal track, was tested by using intestine epithelial cells. It was found that L. acidophilus 107A had a wider anti-microbial spectrum and a superior adherence against intestinal track than other microbials.
In selection of coating materials, analysis of processability, optimization of coating process, comparison of physical properties, and possibility of prebiotics as coating materials were studied to develope a new powder surface modification technology of probiotics for improvement of stability and functionality as a feed additive in swine diets.
Five enteric coating materials were compared as a cadidate coating material of powdered probiotics. Among 5 materials, Sureteric and CompritoI revealed a relatively good and smooth coating ability when the ratio of probiotic and coaling material was 9:1 at 15,000 rpm for 3 min by Hybridization System. Some coating materials needed additional grinding after particle size analysis. Prebiotics which are known to help or promote health benefit bacteria in large intestine. was not perfect as a single coating material But after the secondary coating with Sureteric, a
significant improvement of coating quality was observed.
Viable cell counts of a powder surface-modified probiotic remained viable well.
And acid tolerance of probiotic was very high even when modification process was absent. possibly due to the high acid tolerance of probiotic itself. And the heat stability has much improved by powder surface modification process.
Finally. L. acidophilus ATCC 107A was selected as probiotic for feed additive in swine diets at 0.2% level. The stability of different forms of additive was compared between pellet type and mesh type at various storage temperatures. For pellet type, initial viable cell count has decreased by heal and pressure shock during pellet processing, drying, contact of water, and possible contamination. According to the viable cell count analysis, mesh type was more reliable than pellet type to ensure stability of probiotic.
The effects of probiotics and coated-probiotics on growth performance of pigs were investigated. Four strains of microbials were selected based on in vitro experiments. These microbials were used as probiotic in weaned pigs diets. They had a growth promoting effects when compared with control group. The Lactobacillus acidophilus 393 and Lactobacillus acidophilus 107A fed pigs showed more higher average daily gains among the pigs fed probiotics. Feed efficiency was also improved. The diarrhea frequency was also reduced in pigs fed probiotics compared to non probiotics fed groups. This result showed that probiotics had a positive effect on intestinal ecosystem. The Lactobacillus acidophilus 107A and lactobacillus acidophilus 393 had a positive effect on growth performance. They were used Lo investigate the synergy effects with prebiotics. The synergy effect of Lactobacillus acidophilus 107 A and lactobacillus acidophilus 393 with lactulose was superior to other prebiotics in vitro experiments. The lactulose was used as a prebiotics. Prebiotic improved the growth promoting effects of lactobacillus acidophilus 393. However, the synergy effects of Lactobacillus acidophilus 107 A with prebiotics was not observed. The average daily gains of the pigs fed Lactobacillus acidophilus 107 A and the pigs fed Lactobacillus acidophilus 309 with prebiotics were not different. However, the feed efficiency of the pigs fed Lactobacillus acidophilus 107A was higher than that of the pigs fed Lactobacillus acidophilus 309 with lactulose. Finally, Lactobacillus acidophilus 107 A was selected as a candidate microbial as probiotics. The Lactobacillus acidophilus 107A was used as probiotics for hybridization with coating material. The Lactobacillus acidophilus 107A and encapsulated Lactobacillus acidophilus 107 A were used as probiotics in growing-finishing pigs. The Lactobacillus acidophilus 107A and encapsulated Lactobacillus acidophilus 107A had a same effects as antibiotics was added to feed. Average daily gains of pigs fed Lactobacillus acidophilus 107A and encapsulated Lactobacillus acidophilus 107A as a probiotics did not differ from that of pigs fed antibiotics. And there were no difference among treatments in meat quality.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 3
• SUMMARY ... 9
• CONTENTS ... 12
• 목차 ... 14
• 제1장 연구개발 과제의 개요 ... 17
• 제1절 연구개발의 목적 ... 17
• 제2절 연구 개발의 필요성 ... 17
• 제1항 기술적 측면 ... 19
• 제2항 경제ㆍ산업적 측면 ... 20
• 제3항 사회문화적 측면 ... 20
• 제3절 연구 개발의 범위 ... 22
• 제1항 기능성 균주와 prebiotics를 이용한 복합 생균제 개발 ... 22
• 제2항 분체 복합화 기술 확립 ... 22
• 제3항 복합생균제의 사료 안정성 및 동물 능력개선효과 검증 ... 22
• 제2장 국내외 기술개발 현황 ... 23
• 제3장 연구개발 수행 내용 및 결과 ... 27
• 제1절 기능성 균주와 Prebiotics를 이용한 복합생균제 개발 ... 27
• 제1항 실험재료 및 방법 ... 27
• 1. 유산균주의 분리, 동정 및 보존 ... 27
• 2. Probiotics의 선별 ... 27
• 3. Prebiotics의 선별 ... 28
• 4. 생균제용 균주의 최적 성장 배지 조성 탐색 ... 30
• 5. 최적 생산 및 회수조건 설정을 위한 균주의 농축, 동결 및 산소 저항성 조사 ... 32
• 6. Prebiotics의 최적 첨가비율 설정 ... 32
• 7. Prebiotics의 상호 시너지 효과 검증 ... 32
• 8. 선발된 균주의 최적 건조 및 저장방법 조사 ... 35
• 9. 생균제의 저장기간 중 활력변화 비교 분석 ... 35
• 10. 유전자 수준에서의 생균제의 동정 ... 35
• 11. 다양한 병원성균주에 대한 박테리오신의 항균능력 조사 ... 38
• 12. 장관 세포에 점착성 ... 38
• 제2항 실험결과 및 고찰 ... 39
• 1. Probiotics의 선별 ... 39
• 2. Prebiotics의 선별 ... 45
• 3. 최적 생산 배지 조성의 설정 ... 50
• 4. 농축, 동결, 산소 저항성 균주의 선별 ... 61
• 5. 동물실험용 생균제의 선별 및 생산 ... 63
• 6. Prebiotics 및 probiotics의 선발 ... 63
• 7. Prebiotics의 최적 첨가비율 설정 ... 65
• 8. Prebiotics의 최적 혼합비율 설정 ... 70
• 9. 선발된 균주의 최적 건조 및 저장방법 조사 ... 77
• 10. 생균제의 저장기간 중 활력변화 비교 분석 ... 81
• 11. 유전자 수준에서의 선발균주의 동정 ... 84
• 12. 선발균주의 항균능력 ... 89
• 13. 장관 세포에 대한 점착성 연구 ... 92
• 제2절 분체 복합화 기술 확립(위탁연구과제) ... 94
• 제1항 실험재료 및 방법 ... 94
• 1. 생균제 및 prebiotics의 분체복합화 과정 ... 94
• 제2항 실험결과 및 고찰 ... 101
• 1. 분체복합화 피복재의 screening 및 피복조건 최적화 ... 101
• 2. 가공 및 저장중 생균제의 활력 변화 비교분석 ... 115
• 3. 생균제의 사료첨가형태 및 제품 안정성을 고려하여 분체복합화 생산공정을 scale-up하는 조건 설정 ... 132
• 제3절 복합생균제의 사료안정성 및 동물능력개선효과 검증 ... 141
• 제1항 실험재료 및 방법 ... 141
• 1. 선별된 생균제의 생체실험 및 안정성 평가 ... 141
• 2. 분체복합화된 생균제 첨가물의 안정성과 효능 검증 ... 142
• 3. 실용화 단계 제품의 효과 검증 및 경제성 검증 ... 144
• 제2항 실험 결과 ... 150
• 1. 선별된 생균제의 생체실험 및 안정성 평가 ... 150
• 2. 분체복합화된 생균제 첨가물의 안정성과 효능 검증 ... 157
• 제4장 목표달성도 및 관련분야에의 기여도 ... 175
• 제1절 기능성 균주와 Prebiotics를 이용한 복합생균제 개발 ... 175
• 제2절 분체 복합화 기술 확립 ... 176
• 제3절 복합생균제의 사료 안정성 및 동물 능력개선효과 검증 ... 176
• 제5장 연구개발 계획의 활용계획 ... 179
• 제1절 추가 연구의 필요성 ... 179
• 제2절 타 연구에의 응용 ... 179
• 제6장 연구개발과정에서 수집한 해외과학기술정보 ... 180
• 제7장 참고문헌 ... 181
• 끝페이지 ... 185