cPCR 기법을 이용한 초기배양 pH에 의한 반추위 섬유소 분해 박테리아의 부착 및 발효에 관한 연구

김민석,성하균,김현진,이상석,장종수,하종규,Kim, M.S.,Sung, H.G.,Kim, H.J.,Lee, Sang-S.,Chang, J.S.,Ha, J.K. 한국축산학회 2005 한국축산학회지 Vol.47 No.4

본 연구는 배양초기 pH 조건이 F. succino- genes의 섬유소 부착과 섬유소 소화에 미치는 영향을 보고자 실시하였다. 선정된 specific primer를 이용하여 F. succinogenes의 genomic DNA로부터 445bp의 16S rDNA 절편을 증폭하여 205bp의 internal control을 제작하였고, cPCR 결과로부터 박테리아 수를 계산할 수 있는 표준곡선의 회귀식($r^2$>0.99)을 얻을 수 있었다. In vitro 배양초기 pH에 따른 F. succ- inogenes의 cellulose 부착을 cPCR로 모니터링한 결과, 발효과정 전 기간동안 초기 pH가 6.8과 6.2일 때 cellulose 건물 g당 부착 균주의 수가 pH 5.6일 때 보다 높았으나, pH 6.8과 6.2 사이에서는 큰 차이는 없었다(p>0.05). Cellulose 분해는 배양시간이 진행됨에 따라 증가 되었으며, 분해 정도는 pH가 증가함에 따라 더 높았다. 배양초기 pH가 6.8, 6.2 그리고 5.8일 때 48시간동안 감소한 pH는 각각 0.24, 0.58 그리고 0.16 이었다. 가스 생산량은 발효 시간이 경과함에 따라 pH가 높을수록 더 많았다. 결론적으로 발효 초기 pH는 F. succinogenes에 의한 cellulose 소화, 가스 생산, pH 변화 및 cellulose 부착에 큰 영향을 주었으며, 특히, 낮은 pH(5.8)에서는 섬유소 소화 및 박테리아 부착을 현저한 감소 시켰다. The cPCR technique was used to monitor rumen fermentation and attachment of Fibrobacter succinogenes to cellulose at different pH in the in vitro culture medium. The target fragments of 16S rDNA(445 bp) were amplified from genomic DNA of F. succinogenes with specific primers and internal controls(205 bp) were constructed. Cell counts were estimated from the amounts of genomic DNA, which was calculated from cPCR results. F. succinogenes in pH 6.8 and 6.2 showed apparently higher attachment than in pH 5.8 during all incubation time. There were some difference between pH 6.8 and 6.2 in the degree of attachment, but the different was not significant (P>0.05). Cellulose degradation increased in process of incubation time and the increasing rate was higher when initial pH was higher. The pH in culture medium decreased regardless of initial pH in course of incubation time. After 24 h of incubation, medium pH was dropped by 0.24, 0.58 and 0.16 units from original medium pH 6.8, 6.2 and 5.8, respectively. More gas was produced at higher initial pH in the same manner as in cellulose degradation. In summery, Initial pH of rumen culture in vitro significantly influenced cellulose digestion, gas production, pH change and bacterial attachment. Especially, low pH(5.8) resulted in much lower bacterial attachment and fiber digestion compared to higher medium pH.

청보리 사일리지 첨가 발효사료가 임신돈의 분 성상 및 악취물질 농도에 미치는 효과

조성백,김창현,황옥화,박준철,김동운,성하균,양승학,박규현,최동윤,유용희,Cho, S.B.,Kim, C.H.,Hwang, O.H.,Park, J.C.,Kim, D.W.,Sung, H.G.,Yang, S.H.,Park, K.H.,Choi, D.Y.,Yoo, Y.H. 한국축산환경학회 2011 축산시설환경학회지 Vol.17 No.3

본 연구는 청보리 사일리지가 첨가된 발효사료를 임신돈에 급여하였을 때 분에서의 악취물질 농도와 미생물 변화를 조사하고, 변비예방 효과를 확인하기 위하여 수행되었다. 1. in vitro 시험결과 페놀은 타피오카, 비트펄프, 밀기울, 루핀종실에서 검출되지 않았으며, p-크레졸은 9.62~52.11 mg/L 범위로 검출되었는데, 루핀종실이 가장 높았고, 타피오카가 가장 낮았다. 또한 타피오카와 비트펄프에서는 인돌과 스카톨이 검출되지 않아 악취저감용 원료사료로 타피오카와 비트펄프가 가장 유용한 것으로 평가되었다. 2. 비트펄프를 기초로 배합한 악취저감사료를 28일간 임신돈에 급여한 후 분을 채취하여 성상을 조사한 결과 대조구에 비하여 발효사료 급여구에서 수분함량이 높고 경도가 낮았으며, 대장균 (E. coli) 수는 적었으며, 유산균 (lactobacilus) 수는 많았다. 3. 돼지의 분 중 p-크레졸과 스카톨 농도를 측정한 결과 대조구에서는 각각 33%와 67%가 검출한계 이하로 조사되었으나, 발효사료 구에서는 각각 67%와 100% 검출한계 이하로서 조사되어 발효사료를 섭취한 돼지의 분에서 악취가 감소될 수 있었다. 4. 따라서 임신돈에 청보리사일리지가 첨가된 발효사료를 급여하였을 때 대장의 기능이 개선되어 변비예방과 악취감소의 효과를 기대할 수 있을 것이다. The study was conducted to investigate the effect of feeding fermented diet including whole crop barley silage on the odor reduction and microbial population change in feces, and the constipation prevention of pregnant sows. The concentration of phenol was not detected in tapioca, beet pulp, wheat bran and lupin seeds, while that of p-cresol was ranged between 9.62 and 52.11 mg/L showing that lupin was highest and tapioca was lowest. It was determined that tapioca and beet pulp were useful feed ingredients to reduce odor due to their lower contents of phenol and indole compounds. Ten pregnant sows were allocated to control group and fermented diet group in 5 sows in each group. They were fed 3.0 kg DM/d of diets for 28 days. Feces was examined and showed that the feces from the fermented diet group was observed with the higher moisture content and the lower hardness than that of the control diet group and the population of E. coli was decreased and the population of lactobacilus was higher than that of the control diet group. The concentrations of p-cresol and skatole were lower than the detection levels at 33% and 67% among the samples of feces of the control group and at 67% and 100% among the samples of the feces of fermented diet group respectively. Thus it is expected that the odor from the feces of pregnant sows fed the fermented diet could be reduced compared with that of control group. Therefore, it is suggested that feeding fermented whole barley diet to pregnant sows improve the function of intestine and reduce the rate of occurrences of constipation and odor levels.

비육돈 사료의 고추냉이 첨가수준별 분뇨의 악취물질 농도 변화

이강훈,황옥화,박규현,양승학,송준익,전중환,이준엽,오상집,성하균,최동윤,조성백,Lee, K.H.,Hwang, O.H.,Park, K.H.,Yang, S.H.,Song, J.I.,Jeon, J.H.,Lee, J.Y.,Ohh, S.J.,Sung, H.G.,Choi, D.Y.,Cho, S.B. 한국축산환경학회 2012 축산시설환경학회지 Vol.18 No.suppl.

서양고추냉이를 비육돈에 급여하여 분뇨에서 발생되는 악취물질의 농도변화를 조사하였다. 1. 서양고추냉이 분말을 사료에 혼합하여 돼지에 급여한 후 분뇨의 단쇄지방산 농도를 측정한 결과, 사료에 고추냉이를 0.02% 혼합했을 때 아세트산, 프로피온산, 부티르산, 이소부티르산, 이소발레르산, SCFA 및 I-SCFA 농도가 가장 높았다(p<0.05). 2. 페놀과 p-크레졸을 합한 페놀류 농도는 고추냉이를 0.01~0.02% 첨가하였을 때 대조구보다 감소하였다(p<0.05). 인돌과 스카톨을 합한 인돌류 농도는 사료에 고추냉이를 0.03% 첨가하였을 때 가장 낮았다(p<0.05). 3. 따라서 분뇨의 페놀류 농도를 줄이기 위해서는 사료에 고추냉이를 0.02% 이하로 첨가하고, 인돌류 농도를 줄이기 위해서는 사료에 고추냉이를 0.03% 정도 첨가하여야 할 것으로 판단된다. This study was performed to test the effect of horseradish powder in fattening pig diet on odorous compound concentration from manure. Twenty fattening boars [Landrace ${\times}$ Yorkshire) ${\times}$ Duroc] weighing an average body weight of $68.4{\pm}4.95}$ kg were randomly assigned to one of 4 treatments (horseradish level in diets: 0, 0.01, 0.02, 0.03%) based on their body weight. This experiment was a randomized complete block (RCB) design using 5 pigs per treatment with 1 pig per metabolizable cage. Pigs were fed experimental diet (amount proportional to 3% of their body weight) twice a day (09:00 and 16:00) for 7 d after having 14 d adaptation period. Experimental diets were mixed with water by 1:2.5 v/v. Concentration of volatile fatty acids (VFAs) was highest (p<0.05) when pigs fed diet with 0.02% horseradish powder. Level of phenol compounds including phenol and p-cresol were decreased (p<0.05) in 0.01~0.02% horseradish treatment group compared with control group. Concentration of indoles including indole and skatole was lowest (p<0.05) in 0.03% horseradish treatment group compare to others. Therefore, results from our current study suggest that the optimal levels of horseradish powder in diet for reducing phenol and indole compounds in pig manure were 0.02 and 0.03%, respectively.

반추가축영양 : 사료 단백질의 Fraction과 In situ 단백질 분해율의 상관관계에 관한 연구

이세영 ( S. Y. Lee ),정유석 ( Y. S. Chung ),송재용 ( J. Y. Song ),박성호 ( S. H. Park ),성하균 ( H. G. Sung ),김현진 ( H. J. Kim ),고종열 ( J. Y. Ko ),하종규 ( J. K. Ha ) 한국동물자원과학회 ( 구 한국축산학회 ) 2007 한국축산학회지 Vol.49 No.3

녹용분말 급여가 성장기별 흰쥐의 성장 및 장기발육에 미치는 효과

성하균,김동균,신형태 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.5

The velvet antler of Korean sika deer has been used to the important resources for human health care with ginseng in Korea and Chinese. For studying on biological function of deer velvet being recorded in many ancient literatures, this experiment was conducted to evaluate the effects of powdered velvet antler on growth, feed intake, feed efficiency and intestinal organ development in growing and adult Sprague-Dawley Rats. Experiments were designed by adding amount of powdered velvet antler such as control (non-supplementation), powdered velvet antler Ⅰ (PVA Ⅰ, recommended dos) and powdered velvet antler Ⅱ (PVA Ⅱ, thrice recommended dose). The recommended dose of powdered velvet in this experiment was calculated with metabolic body weight of rats, which based on the recommended amounts for 70㎏ of human. The growths of growing and adult rats generally appeared advantage in PVA supplementation. The final body weight of control, PVA Ⅰ and Ⅱ were respectively 470±39.00g, 478±3033g and 475±22.72g in growing rats, and 485±38.50g, 521±38.67g and 508±34.44g in adult rats. The average daily feed intake were not significantly influenced but the feed efficiency ratios (feed/gain) were improved by PVA supplementation. The feed/gain ratios of control, PVA Ⅰ and Ⅱ were respectively 5.99, 5.47 and 5.54 in growing rats, and 9.04m 7.73 and 8.18 in adult rats. In case of developments of liver, heart, kidney and stomach, we obtained favorable results in both PAC Ⅰ and Ⅱ, but their results were not significantly different according to amount of PVA supplementation. Because liver and heart are important organs in the circulation of blood, their favorable effects suggest that velvet antler may have factors contributing hematopoiesis. Conclusively, supplementation of that velvet antler may have factors contributing hematopoiesis. Conclusively, supplementation of powdered velvet antler resulted in an improved growth, feed efficiency and development of some intestinal organs in growing and adult rat. Even if further investigation of dose in human health care are should be performed, this experiment appeared the best desirable results in PVA Ⅰ, recommended dose of powdered velvet antler.

유형별 완전혼합사료 급여가 반추위내의 발효성상 및 영양소 소화율에 미치는 영향

이덕윤,고종렬,최낙진,이상석,송재용,이세영,박성호,성하균,하종규 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.5

This study was conducted to examine effects of feeding dry TMR(DTMR), wet TMR(WTMR) and fermented TMR(FTMR) on rumen fermentation, enzyme activity and digestibility in the total tract of sheep. Three rumen cannulated sheep were used in a 3 × 3 latin square design. The present results showed that ?, NH_(3)-N, total and individual VFA, A/P ratio and enzymes (CMCase, Xylanase and Protease) activity in the rumen were higher in WTMR and FTMR compared with DTMR. In addition, dry matter, organic matter, crude protein, ether extract, NDF and ADF digestibility in the total tract were also higher in WTMR and FTMR compared with DTMR. Therefore, the present results showed that WTMR and FTMr are better than DTMR for rumen fermentation and nutrients digestibility.