This in vitro study investigated the enhancement of rumen bacterial adhesion on a substrate to increase the digestibility of rice straw in Hanwoo cattle. The rice straw was pretreated with enzymes, probiotics, or ammonia, and the effects on the enhancement of bacterial adhesion and fiber degradation were analyzed using in vitro rumen fermentation. Enzyme treatment included spraying of cellulase and xylanase at 40 and 120 U per g of rice straw, respectively; Saccharomyces cerevisiae culture of 1.0×107 CFU was sprayed as a probiotic treatment per gram of rice straw; ammonia was sprayed at 0.3% per gram of rice straw. Following enzyme treatment, Fibrobacter succinogenes formed a higher adhesion colony than the control group (7.26±0.03 and 8.43±0.20) after 6h and 12h of in vitro culture (p<0.05), respectively. Attachment of Ruminococcus flavefaciens also increased following enzyme treatment (p<0.05) after 6 and 12 h compared to that of the control (5.18±0.06 and 6.60±0.15); and R. albus attachment showed a significant increase compared to that of the control (5.94±0.15) after 6 h of incubation (p<0.05). Probiotic treatment increased attachment of F. succinogenes in comparison with untreated rice straw after 6 h and 12 h of fermentation (p<0.05); R. flavefaciens attachment showed an increase only after 6 h of culture (p<0.05); R. albus was not affected. Attachment of F. succinogenes, R. flavefaciens, and R. albus increased with ammonia treatment after 6 h and 12 h (p<0.05). Dry matter digestibility was higher after the enzyme treatment (3.45±0.21 and 7.04±0.09) than in the control group(1.85±0.08 and 3.94±0.04) after 6 and 12 h of in vitro culture (p<0.05), respectively. It was also higher than that of untreated rice straw after probiotic and ammonia treatments (p<0.05). There was an increase in the enhancement levels of bacterial adhesion depending on the type of fibrolytic bacteria following enzyme, probiotic, and ammonia treatment. These treatments improved digestibility. This enhancement is considered to be greater following enzyme and ammonia treatments than with probiotics.

This study was to research the relationships between rice straw degradation and changes of fibrolytic bacteria population during the in vitro rumen fermentation. Dry matter(DM) digestion of rice straw and population of fibrolytic bacteria were measured at the 0. 4, 8, 12 and 48 hours during the incubation. The populations of F. succinogenes. R. albus and R. flavefaciens were defined as log copy number of 16S rDNA by technical method of Quantitative real-time PCR. Total population of F. succinogenes, R. flavefaciens and R. albus was sum of bactera attached on rice straw and suspended in medium. It's population was increased with incubation, reached top level of 29.0 Log copy No at the 24 hour and then decreased. In the meantime, DM digestion of rice straw showed the higher increasement from the 8 hour to the 24 hour than from the 0 hour to the 8 hour, and then a slowdown in increasing trend of digestibility. Attachments of F. succinogenes, R. flavefaciens and R. albus were detected immediately after start of in vitro rumen incubation. At the same time, the colonized bacterial share were respectively 34.5%, 84.4% and 67.9% in total population. All of them was reached the highest colonized bacterial share above 94.7% at the 4 hour incubation. However population of attached bacteria was shown the highest level at the 12 hour or the 24 hour incubation. Kinetics of colonization were formed area of top speed from the 12 hour to the 24 hour and respectively reached 10.33, 9.28 및 8.30 Log copy No/h/g DM at the 24 hour by F. succinogenes, R. flavefaciens and R. albus. The kinetics of rice straw degradation was formed top level of 0.95% DM/h at the 24 hour. The present results gave clear evidence that degradation of rice straw was increased with the development of total fibrolytic bacteria in process of rumen fermentation. Also, their attachment was largely occurred immediately after insertion of rice straw, the colonized bacteria was actively proliferated, and then degradation of rice straw was maximized.

The present in vitro study was conducted to examine the effect of buffer solubility of eight protein feeds (coconut meal, distillers grain, sesame meal, perilla meal, soy source cake, rape seed meal, soybean meal and lupine) on the fermentation characteristics, degradability of dry matter (DM) and crude protein (CP), and methane (CH4) production by rumen microbes. Buffer extraction increased pH (P<0.05 ~ p<0.001) of the culture solution but tended to lower ammonia- N concentration for all protein feeds. Total volatile fatty acids(VFAs) and each VFAs concentrations in all incubation was decreased by buffer extraction (P<0.01 ~ P<0.001). Also, molar proportion of acetate in 1h (P<0.001), 3h (P<0.01) and 12h (P<0.05) incubations and molar proportion of propionate in 1h (P<0.001), 3h (P<0.01), 6h (P<0.05) and 12h (P<0.05) were decreased by buffer extraction. But molar proportion of butyrate in 1h (P<0.001), 3h (P<0.01) and 6h (P<0.05) were increased by buffer extraction. The in vitro effective degradability of dry matter (P<0.001) and CP (P<0.001) was decreased by buffer extraction. The methane production (P<0.01~P<0.001) in all incubation was decreased by buffer extraction. The results from in the current study might be useful for diet formulation to improve the feed efficiency of the ruminant animals without massive loss of major nutrients.

This study was conducted to examine the effects of defaunation (removal of live protozoa) on fermentation characteristics, degradation of ryegrass hay and CH4 (methane) production by rumen microbes when incubated with plant oils (SO, sunflower oil and LO, linseed oil) in vitro. Sodium lauryl sulfate (0.000375 g/ml) as a defaunation reagent was added into the culture solution and incubated anaerobically up to 24 h at 39℃. pH from defaunation was increased for all treatments from 6 h incubation times (p<0.01-0.001) compared with those from fauantion. Concentration of ammonia-N from defaunation is higher than that from faunation at 3 h (p<0.001), 12 h (p<0.05) and 24 h (p<0.001) incubation times. Defaunation decreased (p<0.01-0.001) total volatile fatty acid concentration at all incubation times. Molar proportions of C2 (acetate, p<0.05-0.001) and butyrate (p<0.01-0.001) were also decreased by defaunation at all incubation times. Molar proportion of C3 (propionate), however, was increased by defaunation at all incubation times (p<0.001). Thus the rate of C2 to C3 was decreased by defaunation at all incubation times (p<0.001). Defaunation decreased ED (effective degradability) of dry matter (p<0.001) and ED of neutral detergent fiber (p<0.001) of ryegrass hay. Defaunation decreased total gas, CH4 production, CH4 % in total gas and CH4/CO2 at all incubation times (p<0.001). Oil supplementation decreased total gas (p<0.05-0.001), CH4 production (p<0.001) and CH4 % in total gas (p<0.001) compared with control at all incubation times. The result of this study showed that defaunation combined with oil supplementation may cause an alteration of microbial communities and further medicate the fermentation pattern, resulting in both reduction of degradation of ryegrass hay and CH4 production. No difference, however, was observed in all the examinations between SO and LO.

본 연구에서는 재고미의 사료가치 평가를 위해 무처리(Control)를 비롯한 Extruding, Roasting, Steam 처리구들의 영양소 성분분석, in vitro 소화율, in situ 소화율을 분석하였다. In vitro 실험에서 Extruding은 건물 소화율과 gas 발생량이 타 처리구에 비해 상대적으로 높았고, 가장 낮은 pH를 보였으며, 특히 배양 시간 초기에 소화가 신속히 발생하였다. NH3-N 함량은 배양 6 h 이후에서 Control이 Extruding, Roasting, Steam에 비해 유의적으로 높은 값을 보였고, 이는 가열처리된 단백질의 반추위 미생물의 이용성 저하에 따른 결과로 보이나, 더 정확한 결과를 위해 건물 소화율 분석뿐 아니라 CP 소화율 분석이 필요할 것으로 사료된다. VFA 발생량에서는 Extruding이 다른 처리구들에 비해 배양 6, 12 h에서 유의적으로 높았는데 이는 Extruding 처리구의 분해가 배양 초기에 주로 이어져 발생한 결과로 추정된다. 전분을 발효시키는 박테리아에 의해 높은 비율로 생산되는 propionate 증가 폭에서도 같은 경향을 보였다. In situ 소화율은 in vitro의 소화율과는 다소 차이가 있는 것으로 여겨지나, 이는 실험 방법에 대한 오차로 여겨질 수 있으며, ED(유효분해율)를 통해 in vitro 시험과 소화율의 경향성이 유사한 것을 알 수 있었다. 따라서 본 연구결과를 근거로 상대적으로 적합한 사료가공 처리방법은 무처리, Roasting 및 Steaming인 것으로 보여진다. 또한 무처리(control)의 경우 가공처리에 따른 추가비용 발생이 없으므로 경제적으로 유리할 것으로 판단된다. 재고미가 TMR의 원료로 이용될 수 있다면 국내에서 자급 가능한 사료원료로써 우수한 경제성을 지닐 수 있을 것이다.