This study was conducted to estimate the effects of the forage process on rumen fermentation characteristics and greenhouse gas emissions of rye. Rye was grown at the Taeyoung Livestock farm and harvested at the heading stage. The harvested rye (5 kg) was sub-sampled for fresh forage, hay, and silage in triplicates. The sub-sampled rye was freeze-dried or air-dried for fresh forage or rye hay, respectively. For rye silage, the sub-sampled rye forage was ensiled into a 10 L mini bucket silo and stored for 90 days. For 72 h rumen incubation, each forage (0.3 g) was placed into the incubation bottle with the rumen mixture (30 mL) in quadruplicates. After the incubation, total gas was measured and sub-sampled for CO2 and CH4 analyses, and the bottle content was centrifuged for in vitro digestibilities of dry matter (IVDMD) and neutral detergent fiber (IVNDFD), and rumen fermentation characteristics. Silage had higher crude protein, crude ash, and acid detergent fiber concentrations than fresh forage and hay but lower non-fiber carbohydrates and relative feed value (p<0.05). And, silage had higher lactic acid bacteria than the other forages but lower pH (p<0.05). After 72 h incubation in the rumen, fresh forage had higher IVDMD and butyrate content than the other forages (p<0.05). However, silage had higher rumen pH and propionate content than the other forages but lower A:P ratio (p<0.05). Regarding greenhouse gases, silage had lowest total gas (mL/g DMD and NDFD) and CH4 (mL/g DMD and NDFD) emissions, while fresh forage had lowest CO2 (mL/g DMD) emission (p<0.05). Therefore, this study concluded that the ensiling process of rye can effectively mitigate greenhouse gas emissions of Hanwoo.

The study aimed to investigate the impact of varying levels of neutral detergent-soluble fiber (NDSF) in Hanwoo growing cattle diets on rumen fermentation and methane (CH4) emissions. An in vitro rumen fermentation experiment utilized feeds with different NDSF levels, incorporating ingredients such as corn grain, soybean meal, soybean hulls, palm kernel meal, beet pulp and timothy hay. The NDSF contents in the diets were 9.02% (T1), 10.09% (T2), 12.42% (T3) and 14.63% (T4). In vitro dry matter digestibility (IVDMD) at 48 h was 7.4% higher for T4 compared to T1 (p<0.05). Total gas production at 48 h was 6.6% higher for T4 than T1 (p<0.05). CH4 production significantly decreased at 9 h and 12 h for T1 and T2 (p<0.05). At 48 h, CH4 production was 5.6% higher for T4 compared to T1 and 6.7% higher compared to T2 (p<0.05). At 12 h ans 24 h, the ammonia nitrogen concentration of T4 was approximately 33.1% and 40.4% lower, respectively, compared to T1 (p<0.05). The acetate to propionate ratio at 48 h was approximately 18.8% higher for T4 than T1 (p<0.05). From 9 h to 48 h, the proportions of butyrate and valerate were significantly higher for T4 (p<0.05). At 48 h, the dominant phylum in T4's rumen microbial community was Candidatus Thermoplasmatota Methanomassiliicoccus, an Archaea. Therefore, this study confirmed that increasing the NDSF content in growing Hanwoo cattle diets up to 12.42% increases IVDMD without increasing CH4 emissions, which is expected to positively impact Hanwoo productivity.

본 실험에서는 외인성 효소 첨가제 및 혼합 세균 배양을 통한 고상발효(Solid-state fermentation, SSF)가 채종박(Rapeseed meal, RSM)의 체외건물소화율(In vitro dry matter digestibility, IVDMD) 및 단쇄지방산(Short chain fatty acid) 생성에 미치는 영향을 조사하기 위해 수행되었다. 외인성 효소 칵테일(첨가 및 미첨가) 및 RSM에 대한 SSF(발효 및 비발효)를 나타내는 2 x 2 요인 설계가 적용되었다. 3-step 돼지 소화율 모델을 적용하여 채종박의 건물 소화율을 분석하였으며, 72시간 대장발효 후 상층액을 수집하여 단쇄지방산 생성량을 분석한 후 칼로리 단위로 변환하여 가소화에너지 소화율을 분석하였다. 소장 (IVDMDh) 및 전장 (IVDMDt) 건물소화율에서는 고상발효된 채종박이 더 높게 나타났다 (각각 p < 0.01). 마찬가지로, 외인성 효소 첨가제 처리구에서 채종박의 소장 소화율(IVDMDh)이 증가하는 경향을 나타냈다(p = 0.06). Acetic acid 및 butyric acid의 생산은 대조구에 비해 고상발효 처리 시 유의하게 더 생산되었으며 (각각 p < 0.01), 이는 총 단쇄지방산의 생산 증가 경향을 나타냈다(p = 0.09). 에너지 소화율에서는 채종박의 고상발효 및 외인성 효소제 첨가가 유의적으로 높게 나타났다 ( p < 0.01). 그러므로 채종박의 고상발효 처리는 단백질 이용성을 비롯한 영양적 가치를 향상시키는데 효과적이라고 사료된다.

This study evaluated the effect of lactic acid bacteria (LAB, a mixture of Enterococcus faecium and Lactobacillus plantarum) supplementation, the storage temperature, and storage period on the fermentation characteristics and in vitro ruminal digestibility of a total mixed ration (TMR). The TMR was prepared into two groups, namely, CON (control TMR without the LAB) and ML (supplementing a mixture of E. faecium and L. plantarum in the ratio of 1% and 2% (v/w), respectively). Both groups were divided and stored at 4°C or 25°C for 3, 7, and 14 d fermentation periods. Supplementing LAB to the TMR did not affect the chemical composition of TMR except for the lactate and acetate concentration. Storage temperatures affected (p<0.05) the chemical composition of the TMR, including pH, lactate, and acetate contents. The chemical composition of TMR was also affected (p<0.05) by the storage period. During in vitro rumen fermentation study, the ML treatment showed lower (p<0.05) dry matter digestibility at 24 h incubation with a higher pH compared to the CON. There was no difference in the in vitro dry matter digestibility (IVDMD) of TMR between the CON and ML treatment however, at 24 h, ML treatment showed lower (p<0.05) IVDMD with a higher pH compared to the CON. The effects of storage temperature and period on IVDMD were not apparent at 24 h incubation. In an in vivo study using Holstein steers, supplementing LAB to the basal TMR for 60 d did not differ in the final body weight and average daily gain. Likewise, the fecal microbiota did not differ between CON and ML. However, the TMR used for the present study did include a commercial yeast in CON, whereas ML did not; therefore, results were, to some extent, compromised in examining the effect of LAB. In conclusion, storage temperature and period significantly affected the TMR quality, increasing acetate and lactate concentration. However, the actual effects of LAB supplementation were equivocal.

본 연구의 목표는 S. boulardii 유래 세포벽 추출물 (CWSB)의 첨가가 반추위 발효에 미치는 영향을 평가하는 것이다. 본 연구의 in vitro 발효 실험은 CWSB의 첨가용 량이 다른 2개의 실험으로 구성되었다. 실험 1에서는 대 조구(CON, 시험사료)와 CWSB0.05(시험사료+CWSB 0.5g/kg 첨가), CWSB0.075(시험사료+CWSB 0.75g/kg 첨 가)의 처리구로 구성되었고, 실험 2의 처리구는 대조구 (CON, 시험사료)와 CWSB0.5(시험사료+ CWSB 5g/kg 첨 가), CWSB1.0(시험사료+CWSB 10g/kg 첨가)의 처리구로 구성되었다. In vitro 배양 이후 3, 6, 12, 24, 48h 동안 발 생한 가스 발생량 및 48h 배양 이후 발효 성상을 분석하 였다. 실험 1에서는 CWSB0.05 처리구에서 유의적으로 높 았던 Vmax(p<0.01)를 제외한 모든 반추위 발효성상에서 처 리구간 유의적 차이가 관찰되지 않았고, 이에 따라 사료 내 0.5%, 1%의 고용량 첨가 수준을 설정한 실험 2를 수행 하였다. 실험 2에서의 CWSB1.0 처리구는 대조구에 비해 총 VFA 발생량이 유의적으로 높았으나 전체 배양시간 내 가스 발생량 및 건물 소화율, 섬유소 소화율은 대조구와 유의적인 차이를 나타내지 않았다. 본 연구 결과를 종합 해 볼 때, CWSB의 첨가수준은 0.5% 이상에서 반추위 발 효 개선효과를 나타낼 것으로 추정된다.

반추위 미생물체 단백질의 아미노산 조성이 한우의 에 너지원 사료에 따라 차이가 있는지를 조사하고자 본 연구 가 수행되었다. 본 실험은 국내 한우 비육우 사육에 주로 이용되는 에너지원 사료인 옥수수(T1), 생미강(T2), 소맥 (T3), 소맥피(T4)를 이용하여 in vitro 반추위 발효실험을 진행하여 발효 24시간 후 배양액의 발효 성상, 미생물체 단백질 합성량(MPS), 미생물의 아미노산 조성 및 미생물 균총 변화를 분석하였다. 배양액의 pH는 T4가 6.02로 다 른 처리구들에 비해서 가장 높았으며, T1은 4.84로 가장 낮았다(p<0.05). Total VFA 농도는 T2가 96.80mM로 다른 처리구들에 비해서 가장 높았고, T1이 53.35mM로 가장 낮았다(p<0.05). 미생물체 단백질 합성량은 T3가 222.50g/L로 다른 처리구들에 비해서 높았으며(p<0.05), T1과 T4는 유의적인 차이는 없었지만 T1이 가장 낮았다. DGGE band 분석결과 모든 처리구는 57.5% 이하의 유사도를 나타내었으며, 급여되는 사료에 따라서 미생물 군집 이 다르게 나타났다. 미생물 내 총 아미노산 함량은 T3와 T4가 다른 처리구보다 높았다(p<0.05). 미생물의 아미노산 조성 중 대부분의 필수 아미노산은 T4가 다른 처리구들에 비해서 낮았다. 하지만 methionine은 다른 필수 아미노산 과 다르게 T4에서 가장 높았다(p<0.05). 곡류 사료 위주의 급여 시 결핍되기 쉬운 lysine은 모든 처리구에서 미생물 의 아미노산 조성이 8.05~8.98%의 범위에 있었다. 따라서, 본 연구에서 반추 가축 급여 에너지 사료원에 따른 반추 위 내 미생물 발효특성과 미생물 군집의 차이가 미생물의 아미노산 조성을 변화시킨다는 것을 확인하였다.

The present study was conducted to examine the effect of soybean silage as a crude protein supplement for corn silage in the diet of Hanwoo steers. The first experiment was conducted to evaluate the effect of replacing corn silage with soybean silage at different levels on rumen fermentation characteristics in vitro. Commercially-purchased corn silage was replaced with 0, 4, 8, or 12% of soybean silage. Half gram of the substrate was added to 50 mL of buffer and rumen fluid from Hanwoo cows, and then incubated at 39°C for 0, 3, 6, 12, 24, and 48 h. At 24 h, the pH of the control (corn silage only) was lower (p<0.05) than that of soybeansupplemented silages, and the pH numerically increased along with increasing proportions of soybean silage. Other rumen parameters, including gas production, ammonia nitrogen, and total volatile fatty acids, were variable. However, they tended to increase with increasing proportions of soybean silage. In the second experiment, 60 Hanwoo steers were allocated to one of three dietary treatments, namely, CON (concentrate with Italian ryegrass), CS (concentrate with corn silage), CS4% (concentrate with corn silage and 4% of soybean silage). Animals were offered experimental diets for 110 days during the growing period and then finished with typified beef diets that were commercially available to evaluate the effect of soybean silage on animal performance and meat quality. With the soybean silage, the weight gain and feed efficiency of the animal were more significant than those of the other treatments during the growing period (p<0.05). However, the dietary treatments had little effect on meat quality except for meat color. In conclusion, corn silage mixed with soybean silage even at a lower level provided a greater ruminal environment and animal performances, particularly with increased carcass weight and feed efficiency during growing period.

본 연구는 조사료 원료인 마늘 껍질의 사료적 가치와 메탄 발생량을 조사하기 위해 in vitro 발효 실험을 수행하였다. Garlic husk를 제외하고 국내에서 널리 사용되는 4가지 조사료 원료(Oat hay, Annual ryegrass, Timothy, Tall fescue)를 실험의 처리구로 구성하였다. In vitro 48 시간 발효 후 건물 소화율, 섬유소 소화율, 암모니아태 질소, 휘발성 지방산, pH, 메탄 발생량을 평가하였다. 가스 발생량은 3, 6, 9, 12, 24, 36 및 48 시간에 각각 측정되었다. 최종 가스 발생량에서 Oat hay가 유의적으로 가장 높았다(p<0.01). Garlic husk는 처리구 중 가장 높은 메탄 발생량을 나타내었다(p<0.01). Garlic husk의 건물 소화율은 Oat hay보다 유의적으로 낮았지만 Annual ryegrass, Timothy, Tall fescue보다 높았다(p<0.01). 섬유소 소화율은 Annual ryegrass에서 가장 낮았다(p<0.01). 총 휘발성 지방산 생성량은 Oat hay에서 가장 높았고(p<0.01), Garlic husk, Timothy, Tall fescue는 유사하였다. 본 연구의 in vitro 발효 성상과 메탄 발생량을 고려할 때, Garlic husk는 국내 주요 조사료를 대체할 영양적 가치를 포함하고 있으나 메탄 저감 효과는 미미할 것으로 추정된다.

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.

The objective of this study was to evaluate effects of feeding methods on in vitro ruminal fermentation, total gas and methane production in Hanwoo steers. Six Hanwoo steers fitted with rumen cannula (430 ± 21 kg of body weight) were randomly assigned to one of three feeding systems: 1) feeding forage 1 hour after concentrate, 2) feeding concentrate 1 hour after forage, 3) feeding mixed ration. Rumen fluid sampled from each animals was incubated 24 hours with maize or timothy substrates in in vitro. Ruminal pH was increased in feeding method 2 or maize substrate than that of other methods or timothy substrate (P < 0.001). The production of total volatile fatty acid, acetate, propionate, butyrate, and valerate were increased when steers fed diets using feeding method 1 or rumen fluid was incubated with maize substrate (P < 0.001). Increased production of total gas and methane was observed in feeding method 1 and maize substrate compared to those of other methods or timothy substrate (P < 0.001). Due to the inconsistent results between ruminal fermentation and gas production in this study, further research is required to estimate effects of feeding method on enteric fermentation and gas production in in vivo.

In this study, the effect of forage sources in the total mixed ration (TMR) on in vitro goat rumen fermentation was investigated. Rice straw (RS), Italian ryegrass (IRG), timothy (TIM), and alfalfa (ALF) were used as forage sources. Each forage source was mixed with a commercial goat concentrate diet in the ratio of 1:1. Total 4 TMR were prepared. Rumen simulated in vitro fermentation using goat rumen fluid collected from the slaughterhouse was conducted until 72 th . For fermentation parameters, gas production (GP), volatile fatty acids (VFAs), and ammonia nitrogen (NH3-N) were examined. All assays were performed at 24 th , 48 th , and 72 th h of incubation individually. Contents of crude protein and non-fibrous carbohydrate were greater in the order of RS < IRG < TIM < ALF. Significant treatment effects were found in valerate and NH3-N at 24 th h of incubation (p<0.05). ALF showed the greatest contents of them and RS was the lowest. At 48 th incubation, a significant effect was detected at GP (p<0.05) and RS was greater than others. However, GP of RS was lower than others at 72 th . Significant effects on Total VFA, butyrate, and valerate productions were found at 72 th h of incubation (p<0.05). ALF showed the greatest production. Methane production from all treatments was not significantly different for each incubation time (p>0.05). The present study provided primary information on how goat rumen fermentation responds to different nutrient contents and forage sources of TMR. And the information could be used for the design or optimizing economical diet formulation for goats.

This study was conducted to evaluate the effect of lactic acid bacteria (LAB) inoculation to domestically-cultivated Italian ryegrass (IRG) on silage fermentation and in vitro ruminal fermentation. There were six treatments based on the LAB inoculants: 1) no addition of LAB (negative control: NC), additions of 2) commercially-available LAB (positive control: PC), 3) Lactobacillus plantarum (LPL), 4) L. paracasei (LPA), 5) L. acidophilus (LA), and 6) L. pentosus (LPT). All treatments were inoculated at a concentration of 106 CFU/g and ensiled for 3, 7, 21, and 42 days in triplicate and analyzed for nutritive values when ensiling was terminated. Day 42 silage from all treatments were also examined for in vitro ruminal fermentation. After 42 days, LAB-inoculated silages had higher (P<0.05) lactic acid concentration compared to the NC. In terms of nutritive values, the silages treated with LPA, LA, and LPT showed higher (P<0.05) crude protein and lower (P<0.05) neutral detergent fiber and acid detergent fiber content compared to the rest of the treatment. In vitro ruminal dry matter degradability was not affected by LAB addition. However, LAB-treated IRG had shown higher (P<0.05) ammonia-N compared with that of the NC. LPA had shown the highest (P<0.05) volatile fatty acid concentration among the LAB examined. In conclusion, the addition of a single strain of LAB appeared to produce a quality IRG silage compared with the NC and the PC. Among the strains examined, LPA seemed to be superior to the others.

This study evaluated effects of the addition of garlic powder allicin (GPA) mixture on rumen fermentation with methane in Hanwoo steer. On in vitro trial, two experimental groups were used: control (without GPA) and treatment group (addition of 0.1% GPA mixed with the basal concentrate). Similar to in vitro trial, two experimental groups were used in vivo trial. Five Hanwoo steers (3 steers in one group and 2 in another group; average body weight = 500 ± 43 kg) were assigned by crossover design for 20 d consists of 15 d diet adaptation and 5 d data collection in each experimental period. Daily feed intake and enteric methane production were recorded by an automated head chamber system. The results of in vitro study showed that the GPA treatment group had higher acetic acid (24.30 vs 23.45 mmol/L) and butyric acid (16.55 vs 15.47 mmol/L) concentrations, but lower CH4 production (1.40 vs 2.71 mmol/ml) after 24 h of incubation compared to the control (p<0.05). Total gas, propionic acid, total volatile fatty acid (VFA), and acetic acid: propionic acid ratio were not affected by treatment after 24 h incubation. In the in vivo experiment, rumen pH and VFA were not significantly different between treatments (p>0.05), except acetic acid, which was significantly higher in GPA mixture group (60.97 vs 53.94 mM) than in the control group (p<0.05). Furthermore, no significant differences were recorded in CH4 production (g/d) and CH4 yield (g/kg DMI) between the two groups (p>0.05). In conclusion, the addition of 0.1% GPA mixture reduced CH4 proudcition on in vitro trial, but no effect on in vivo trial.

In in vitro fermentation studies, feed samples can either be included in the in vitro rumen medium using filter bags or can be directly dispersed. The objective of this study was to investigate the effects of different pore sizes of filter bags on the rumen fermentation characteristics in an in vitro system. Corn, soybean meal, and timothy were ground to pass through a 1.0-mm screen and were formulated in the ratio of 70:7:23 based on DM, respectively. The formulated experimental diet (2g/DM) was put in F57 filter bags and R510 nylon bags (Ankom®) which pore sizes were 25 and 50 μm, respectively. An in vitro study was conducted to determine the rumen fermentation characteristics for 3, 6, 12, 24, and 48 h and rumen microbial community at 48 h of incubation. A significantly higher production of gas was observed in the R510 bags than in F57 at all the incubation times (p<0.01). IVDMD (p<0.01) and IVNDFD (p<0.01) were significantly higher, whereas pH (p<0.01) and NH3-N (p<0.01) were lower when R510 bags were used. In the VFA composition, acetate and butyrate were significantly higher (p<0.01) in R510 bags, and propionate and total VFA concentration did not differ (p=0.55 and 0.25, respectively) between F57 and R510 bags. The log copy numbers of bacteria and protozoa did not differ (p=0.69 and 0.94, respectively) between F57 and R510 bags, whereas those of fungi were significantly higher in R510 than in F57 bags (p<0.01). Therefore, the use of R510 may reflect actual rumen fermentation characteristics more precisely than those of F57 because increased gas production, nutrient digestibility and acetate, butyrate proportion were founded in R510.

Fermented total mixed ration (TMR) is a novel feed for ruminants in South Korea. The purpose of this study was to evaluate the effects of lactic acid bacteria (LAB) on the quality of TMR and in vitro ruminal fermentation. Strains of three LAB spp. (Lactobacillus plantarum, L. brevis, L. mucosae) were used in fermentation of TMR. Inoculations with the three LAB spp. lowered pH and increased concentrations of lactic acid, acetic acid, and total organic acid compared to non-LAB inoculated control (only addition of an equivalent amount of water) (p<0.05). Bacterial composition indicated that aerobic bacteria and LAB were higher. However, E. coli were lower in the fermented TMR than those in the control treatment (p<0.05). Among the treatments, L. brevis treatment had the highest concentration of total organic acid without fungus detection. Gas production, pH, and ammonia-nitrogen during ruminal in vitro incubation did not differ throughout incubation. However, ruminal total VFA concentration was higher (p<0.05) in the LAB spp. treatments than the control treatment at 48 hours. Overall, the use of L. brevis as an inoculant for fermentation of high moisture. TMR could inhibit fungi growth and promote lactic fermentation, and enhance digestion in the rumen.

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).

This study was conducted to examine the effect of new inoculants on in vitro digestibility and fermentation characteristics of high moisture rye silage. Rye was harvested at heading stage and divided into 5 treatments, following: No additives(CON); L. plantarum R48-27(NI1); L. buchneri R4-26(NI2); mixture of NI1 and NI2 at 1:1 ratio(MIX); and L. buchneri(LB). The rye forage was ensiled into 10 L bucket silo for 100 days. In vitro digestibility of dry matter and neutral detergent fiber were highest(p<0.05) in NI2 silage. The pH in NI2 and LB silages were lower(p<0.05) than CON silage. Lactate concentration was highest(p<0.05) in NI1 silage. While concentrations of acetate and propionate were highest(p<0.05) in MIX silage. Lactates : acetate ratio was highest(p<0.05) in NI1 silage, but lowest in LB silage. Butyrate concentrations of NI2 and LB silages were lower(p<0.05) than that in CON and NI1 silages. Lactic acid bacteria (LAB) count in all inoculated silages was higher(p<0.05) than that in CON silage, while yeast count in LB silage was lower than in CON, NI1, and MIX silages. In conclusion, application of NI2 inoculant could improve potentially fermentation quality and digestibility of high moisture rye silage.