This study was conducted to evaluate the changes of pH, dry matter digestibility (DMD), NH3-N concentrate, gas production and volatile fatty acid (VFA) through in vitro fermentation by adding horse feces to various juice pomaces fermented with Bacillus, yeast and lactic acid bacteria. The pH range of fermented fluid with juice pomaces was 6.4-7.1, indicating that the digestion by microbial fermentation was normal. Juice pomaces adopted will be helpfully used to assist with digestion by microbes in intestines because approximately 109 CFU/㎖ microbes were grown after 48 hours in fermented fluid. DMD rate gradually increased from 12 hours. It was 39.19% in pomaces of apple, 38.22% in grape, 37.02% in carrot, 36.2% in citrus and 34.35% in mixture respectively after 48 hours. NH3-N concentrate was not changed significantly as it was maintained at 1.5 mg/100㎖ level in the entire treatment group from beginning of fermentation until 12 hours, but increased rapidly from 24 hours. Amount of gas produced was lowest in the mixture and increased rapidly after 12 hours. Total VFA increased from 24 hours and was highest at 48 hours. It was suggested that dry matter digestion was processed while fermented juice pomaces kept proper pH during in vitro digestion, and cellulose degrading microorganisms could act actively in the caecum and colon of horses.

This study was conducted to examine the effect of corn (Zea mays L.) - soybean (Glycine max L.) silage prepared by intercropping method on the nutritive value of the silage, in vitro rumen fermentation characteristics, dry matter degradability, as well as milk yield and milk composition of dairy cows. In a couple of experiments intercropped corn-soybean silage (CSBS) was compared with corn silage (CS) and/or Italian ryegrass hay (IRG). Numerically, CSBS had higher crude protein, ether extract, and lactic acid contents compared to CS. In vitro rumen fermentation analysis demonstrated that up to a 24-h incubation period, both CS and CSBS showed higher total gas production, ammonia N concentration, and dry matter degradability compared to IRG (p<0.05). The investigation on animals was conducted in a commercial dairy farm located in Gyeongju, South Korea, employing 42 Holstein cows that were divided into 2 group treatments: CS and CSBS in a completely randomized design. Although no significant difference was observed in milk yield, animals fed on CSBS showed significantly higher milk protein (p<0.05) and milk fat content (p<0.01), compared to animals fed on CS. Taken together, our findings indicate that corn-soybean silage that is cultivated, harvested, and prepared through intercropping can improve the protein content of the silage, and can also enhance in vitro rumen fermentation, dry matter degradability, and performance of dairy cattle.

본 연구는 팽이버섯 수확 후 배지를 첨가하여 호밀 사일리지 제조 시 팽이버섯 수확 후 배지를 에너 지원으로서 사용하기 위하여 in vitro 반추위 발효실험이 수행되었다. 공시 사일리지는 출수기의 호밀 에 팽이버섯 수확 후 배지 첨가비율(0%, 20%, 40%, 60%)에 따라 제조하여 6주일간 발효시켰다. In vitro 배양액 제조를 위한 반추위액은 농후사료와 볏짚을 40:60의 비율로 급여한 반추위 cannula가 시 술된 Holstein 수소 2두로 부터 채취하였다. In vitro 실험은 발효시간대를 3, 6, 9, 12, 24 및 48시간 으로 설정하고, 각 처리구별로 3반복으로 발효특성과 건물소화율을 측정하였다. In vitro 배양액의 pH 는 발효시간이 길어짐에 따라 낮아지는 경향이었으며, 48시간 경과 시에는 버섯수확 후 배지 60% 첨가 구가 타 처리구에 비해 유의적(p<0.05)으로 낮았다. 미생물 성장율은 배양시간이 경과함에 따라 증가 하는 경향이었으며, 발효 48시간 경과 시에는 버섯수확 후 배지 20% 첨가구가 타 처리구에 비해 유의 적으로(p<0.05) 높았다. Gas발생량은 48시간 발효 시에 대조구가 타 처리구에 비해 유의적(p<0.05)으 로 높았다. 건물소화율은 버섯수확 후 배지의 첨가비율이 높을수록 높았는데, 발효 24시간 및 48시간에 는 R-60구가 처리구 중에서 가장 높았으며(p<0.05), 대조구에서는 전 발효기간 동안 건물소화율이 현 저히 낮은(p<0.05) 상태에 있었다. In vitro 반추위내 발효실험의 결과와 버섯수확 후 배지의 활용성을 고려할 때, 호밀사일리지 제조 시 팽이버섯 수확 후 배지 첨가비율을 원물기준으로 60%수준이 가장 긍 정적인 것으로 나타났다. 향후 대사시험이나 사양시험을 통하여 가축사료로써의 최적 대체 비율을 규명 해야 할 것으로 사료되는 바다.

This study was aimed to evaluate the dose-response the effects of nano-encapsulated conjugated linoleic acids(CLAs) on in vitro ruminal fermentation profiles. A fistulated Holstein cow was used as a donor of rumen fluid. Nano-encapsulated CLAs(LF, 5% of nano-encapsulated CLA-FFA; HF, 10% of nano-encapsulated CLA-FFA; LT, 5% of nano-encapsulated CLA-TG; HT, 10% of nano-encapsulated CLA-TG) were added to the in vitro ruminal fermentation experiment. In the in vitro ruminal incubation test, the total gas production on incubation with nano-encapsulated CLAs was increased significantly according to the incubation time, compared with the control(p<0.05). The tVFA concentrations on addition of LF and HT were significantly higher than that of the control(p<0.05). Thus, nano-encapsulated CLAs might improve the ruminal fermentation characteristics without adverse effects on the incubation process. In addition, the population of Butyrivibrio fibrisolvens which is closely related to ruminal biohydrogenation was increased by adding HT, while decreased by adding LF at 12 h incubation. These results showed that nano-encapsulated CLA-FFA could be applied to enhance CLA levels in ruminants by maintaining the stability of CLA without causing adverse effects on ruminal fermentation profiles considering the optimal dosage.

본 시험은 항염에 효과가 있다고 알려진 식물 추출물의 첨가가 반추위 발효와 메탄 생성에 미치는 영 향을 알아보기 위해 in vitro 실험을 수행하였다. 항염과 항산화 효과가 있는 산뽕나무, 뽕나무, 예덕나 무, 오동나무, 방아풀과 은행나무 6종을 선발하였다. 반추위액과 McDougall buffer 혼합액 15mL과 티 모시 0.3g, 각각의 추출물을 기질의 5%로 넣고 39℃에서 3, 9, 12, 24, 48 그리고 72시간 배양하였다. 항염 효과가 있는 식물 추출물의 첨가는 반추위 발효 성상(pH, 건물소화율, Glucose 농도, 암모니아 농도, 단백질 농도, 미생물 성장량, 휘발성지방산)에는 영향을 미치지 않았다. 총 가스 발생량은 각기 다른 양상을 보였으며, 이산화탄소 발생량은 48시간대에서 예덕나무와 방아풀에서 대조구에 비해 유의 적(p<0.05)으로 높았다. 또한 메탄 발생량은 배양 초기에는 대조구보다 첨가구에서 유의적(p<0.05)으로 감소하였으나, 발효가 진행될수록 대조구에 비해 첨가구에서 더 많은 메탄이 발생하였다. Polyphenol과 flavonoid는 은행나무 추출물구에서 가장 높았다. 본 시험의 결과에서 항염에 효과가 있는 식물 추출물 을 in vitro 반추위 배양액에 첨가하였을 때, 반추위 발효에는 영향을 미치지 않았고, 메탄은 초기 발효 에 저감 효과가 있는 것으로 사료된다.

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 study was conducted to evaluate the effects of microbial culture supplements on ruminal fermentation and fermentative quality of Italian ryegrass silage (IRGS) both in vitro and in situ. Three species of microbes (Lactobacillus casei (LC), Bacillus subtilis (BS), and Saccharomyces cerevisiae (SC)) were used in this study. They were applied to IRGS at 30 days after silage manufacture. Various items were measured using in vitro and in situ incubation technique after each microbial supplement was inoculated into IRGS at 0.5×104 CFU/g. In the first experiment, in vitro ruminal fermentation characteristics of IRGS were evaluated at 0, 12, 24, 48, and 72 hours after microbes were inoculated into IRGS. In the second experiment, in situ fermentation characteristics were investigated at 0, 1, 3, and 5 days after the inoculation of each microbial supplement. In vitro ruminal NH3-N content was significantly (p<0.05) increased in LC-, BS-, and SC-IRGS at 12 hrs post incubation compared to that in control IRGS. In vitro ruminal total VFA concentration and dry matter digestibility (DMD) of IRGS were not significantly difference among LC-, BS-, and SC-IRGS, although they were numerically increased in LC-IRGS than those of the other IRGS. In addition, this study evaluated the fermentation characteristics and in situ DMD of IRGS with the lapse of incubation time up to 5 days. Throughout the incubation times from 1 day to 5 days, the pH value was significantly (p<0.05) lower in BS-, LC-, and SC-IRGS than that in control IRGS. Lactate was significantly (p<0.05) higher, and significantly (p<0.05) butyrate was lower in LC-IRGS than that in other treatments at 0 day. It was higher (p<0.05) in control IRGS than that of BS-, LC-, and SC-IRGS at 1-5 days. In situ DMD tended to increase in BS-, LC-, and SC-IRGS compared to that in control IRGS. Especially, DMD was higher in SC-IRGS than that in other treatments at 0 day. It tended to be higher in LC-IRGS at all incubation time. Taken together, these results suggest that it might be useful to select a microorganism by considering the feeding time of IRGS to ruminants because organic acids and DMD of IRGS were affected by the incubation time of each microorganism with IRG silage, especially for L. casei decreased the content of acetate and butyrate in IRGS.

This study was conducted to evaluate effects of supplementation of Bifidobacterium ruminantium on in vitro ruminal fermentation and methane production. Ruminal fermentation characteristics of Timothy hay (C1), whole barley (C2), Timothy hay + B. ruminantium (T1) and whole barley + B. ruminantium (T2) were evaluated at 0, 3, 6, 9, 12, 24 and 48 h incubation at 39℃. The amount of B. ruminantium culture added into T1 and T2 was 0.3 ml. The pH values ranged from 5.99 to 6.83 in all the treatments. Concentration of NH3-N of C2 and T2 was higher than C1 and T1 at 48 h (p<0.05). The total gas production of C2 and T2 was higher than C1 and T1 at 9 h (p<0.05). The total methane production of treatments with B. ruminantium was not significantly different at 24 and 48 h (p<0.05). Concentration of lactic acid was significantly different between both substrates (p<0.05). Therefore, B. ruminantium supplementation was determined to be insignificant in the in vitro ruminal fermentation and methane production, while a further study was required to investigate relation to lactic acid production with different forage sources.

본 시험은 질산염함유 식물 추출물을 이용하여 in vitro 반추위 발효성상 및 반추위 메탄 발생에 미 치는 영향을 알아보기 위해 수행되었다. 공시재료는 감자, 당근, 배추, 상추 및 시금치 추출물을 사용하 였으며, 발효 기질은 2mm로 분쇄된 timothy 0.3g을 사용하였다. In vitro 실험은 조사료(timothy) 및 배합사료를 6:4의 비율로 급여한 반추위 cannnula가 시술된 한우 암소에서 채취하였다. 반추위액과 McDougall’s buffer를 1:2의 비율로 혼합한 발효액을 0.3g timothy와 식물 추출물이 담긴 50mL serum bottle에 혐기상태로 20mL 분주한 뒤, 39℃에서 9, 12, 24, 48시간 동안 발효하였다. pH는 발 효 시간이 증가함에 따라 점점 감소하며, 6.31~6.96으로 정상범위였다. 건물 소화율은 9시간 배추 추 출물구에서 유의적으로 낮았고, 암모니아 농도는 감자, 배추, 상추 추출물구에서 대조구에 비해 첨가구 에서 유의적으로(p<0.05) 낮았으며, 반추위 미생물 성장량은 발효 24시간대의 당근 추출물구에서 높았 다(p<0.05). Acetate와 propionate 농도는 대조구에 비해 첨가구에서 유의적으로 낮았으며(p<0.05) butyrate 농도는 첨가구간에 발효시간별로 각각 다른 양상을 보였다. 총 가스 발생량은 발효 12시간 및 24시간대에서 배추, 상추, 시금치 추출물구에서 유의적으로(p<0.05) 낮았으며, 메탄 발생량은 감자, 배 추, 시금치 추출물구가 대조구에 비해 유의적으로(p<0.05) 낮았고, 이산화탄소 발생량 또한 첨가구에 비해 대조구에서 유의적으로(p<0.05) 낮았다. 결과적으로 본 시험에 사용된 5종의 질산염 함유 식물 추 출물은 반추위 발효에 악영향을 미치지 않으면서 메탄저감 효과를 나타내었다. 특히 배추 추출물은 반 추위내 메탄발생을 저감시킬 수 있는 식물추출물로서 가능성이 높은 것으로 사료된다.

The objective of this study was to determine the effect of bacterial inoculation (Lactobacillus plantarum or combo inoculant mixed with Lactobacillus plantarum and Lactobacillus buchneri) and addition of fibrolytic enzyme on chemical compositions and fermentation characteristics of whole crop barley (WCB) and triticale (TRT) silage, their ruminal in vitro fermentation, and digestibility. In TRT silage, enzyme addition significantly (p<0.01) decreased NDF content compared to no enzyme addition treatment. Organic acids such as lactate and acetate contents in WCB and TRT silages were significantly (p<0.01) higher compared to those in the control. Particularly, lactate content was the highest in L. plantarum treatment. Fibrolytic enzyme treatment on both silages had relatively higher lactic acid bacteria content, while mold content was lower in both treatments compared to that in the control. In vitro dry matter digestibility was generally improved in WCB silages. It was higher (p<0.01) in TRT with mixed treatment of L. plantarum, L. buchneri, and enzyme compared to others. In vitro ruminal acetate production was relatively higher in treatments with both enzyme and inoculant additions compared to that in the control. Therefore, the quality of silage and rumen fermentation could be improved by inoculants (L. plantarum and L. buchneri) regardless whether whole crop barley (WCB) or triticale (TRT) silage was used. Although it was found that fibrolytic enzyme addition to both silages had various quality and rumen fermentation values, further study is needed

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.

The in vitro experiment was conducted to ensure the supplemental level of spent Flammulina velutipes mushroomsubstrates (SMS) as an energy source in manufacturing of whole crop sorghum silage. Sorghum harvested at heading stage wasensiled with spent mushroom substrates of 20% (S-20), 40% (S-40) and 60% (S-60) as fresh matter basis for 6 week. Theexperiment was conducted by 3, 6, 9, 12, 24, 48 hrs of incubation time with 3 replications. The silages were evaluatedfermentation characteristics and dry matter digestibility (DMD) in vitro. The pH of in vitro solution was inclined to decrease withelapsing the incubation time, and that of the S-20 was significantly (P<0.05) lower than the other treatment at 48 hr ofincubation. Gas production was greater (P<0.05) in the S-20 than the other treatments at 6 and 12 hrs of incubation. Themicrobial growth in vitro was inclined to decrease following 24 hr of incubation, and thereafter sustained the similar levels. Invitro dry matter digestibility (IVDMD) was lowered by increasing the supplemental level of spent mushroom substrate, and was alow level in the S-60 throughout whole incubation time. Althoughthe IVDMD for S-40 was steadily increased from 9 hr ofincubation and reached to similar level with the S-20 at 48 hourof incubation, however SMS for whole crop sorghum silagefermentation might as well add about 20 to 30% in fresh matterbasis when considering DMD.

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.

The objective of this study was to know the in vitro effects of supplemental anthelmintic plant extracts on the inhibition of protozoa for reducing methane production in the rumen. A fistulated Holstein cow was used as a donor of rumen fluid. The plant extracts (Lonicera japonica, Zanthoxylum piperitum, Pyrethrum, Torreya nucifera, Ruta graveolens) known to have anthelmintic effect were added to the in vitro fermentation bottles containing the rumen fluid and medium. The rumen protozoal population was depressed by the addition of Pyrethrum, Torreya nucifera and Ruta graveolens. The methane production was also significantly (p<0.05)reduced by addition of Pyrethrum (2.20 ml/g DM), Torreya nucifera (2.36 ml/g DM) and Ruta graveolens (2.20 ml/g DM). The microbial growth in the treatments of Ruta graveolens or anthoxylum piperitum was the greatest after 12 h and 24 h incubations, respectively. The results of this study indicated that anthelmintic plant extracts appeared to reduce methane production by inhibition of ruminal protozoa related with the methanogens living endosymbiotic in protozoal cells.