Understanding changes in fermentation characteristics and microbial populations of forage silage during ensiling is of interest for improving the nutrient value of the feed for ruminants. This study was conducted to investigate the changes in fermentation characteristics and bacterial communities of whole crop rice (WCR) silage during the ensiling period. The chemical compositions, pH, organic acids and bacterial communities were evaluated at 0, 3, 6, and 12 months after ensiling. The bacterial communities were classified at both the genus and species levels. The dry matter content of WCR silage decreased with the length of storage (p<0.05), but there was no significant difference in crude protein and NDF contents. Following fermentation, the pH level of WCR silage was lower than the initial level. The lactic acid content remained at high levels for 3 to 6 months after ensiling, followed by a sharp decline at 12 months (p<0.05). Before fermentation, the WCR was dominated by Weissella (30.8%) and Pantoea (20.2%). Growth of Lactiplantibacillus plantarum (31.4%) was observed at 3 months after ensiling. At 6 months, there was a decrease in Lactiplantibacillus plantarum (10.2%) and an increase in Levilactobacillus brevis (12.8%), resulting in increased bacteria diversity until that period. The WCR silage was dominated by Lentilactobacillus buchneri (71.2%) and Lacticaseibacillus casei (27.0%) with a sharp reduction in diversity at 12 months. Overall, the WCR silage maintained satisfactory fermentation quality over a 12-month ensiling period. Furthermore, the fermentation characteristics of silage were found to be correlated to bacterial microbiome.

본 연구는 수분함량과 미생물 첨가제가 알팔파 사일리지의 발효특성과 사료가치에 미치는 영향을 알아보기 위해 수행되었다. 알팔파는 개화 10% 시기에서 수확되었으며 수분 함량(M60, M50, M40 및 M30)이 60, 50, 40 및 30%일 때 각각 이용하였다. 1500g을 샘플링한 후, 증류수 10mL을 첨가한 미첨가구(NAD)와 Lactococcus lactis 와 Pediococcus pentosaceus의 혼합물을 증류수(0.1g/10mL)에 1.5 x 1010cfu/g 농도로 희석한 후 접종한 미생물 균주 첨가구(ADD)를 3개월 및 6개월 발효하였다. 수분 및 미생물 첨가제에 따른 연평균 알팔파의 조단백, 중성세제 불용성 섬유 및 산성세제 불용성 섬유에서 차이가 없었다(p>0.05). 모든 발효기간에서 pH는 ADD의 M40에서 가장 낮았다(p<0.05). NAD 처리구에서는 젖산이 M50에서 가장 높았고(p<0.05), ADD 처리군에서는 M40 시험구에서 젖산이 가장 높았다(p<0.05). NAD와 ADD의 M60은 젖산 함량이 다른 수분 함량들에 비해 가장 낮으며(p<0.05) 낙산이 유일하게 검출되었다. 미생물군집의 상대적 풍부도는 ADD 처리구의 M40과 M50에서 Homo LAB (Enterococcus, Lactiplantibacillus, Lacticaseibacillus, Lactococcus, Pediococcus)의 비율이 가장 높았고, Clostridium은 M60에서 가장 높았다.

The aim of this study was to investigate the effect of isolated lactic acid bacteria (LAB) on the quality of high moisture rye silage. Rye forage (Secale cereale L.) was harvested at the heading stage (27.3% of dry matter (DM)) and cut into approximately 3-5 cm lengths. Then, the forage divided into 4 treatments with different inoculants: 1) No additives (CON); 2) Lactobacillus brevis strain 100D8 at a 1.2 x 105 colony-forming unit (cfu)/g of fresh forage (LBR); 3) Leuconostoc holzapfelii strain 5H4 at a 1.0 x 105 cfu/g of fresh forage (LHO); and 4) Mixture of LBR and LHO (1:1 ratio) applied at a 1.0 x 105 cfu/g of fresh forage (MIX). About 3 kg of forage from each treatment was ensiled into a 20 L mini-bucket silo in quadruplicate for 100 days. After silo opening, silage was collected for analyses of chemical compositions, in vitro nutrient digestibilities, fermentation characteristics, and microbial enumerations. The CON silage had the highest concentrations of neutral detergent fiber and acid detergent fiber (p = 0.006; p = 0.008) and a lowest in vitro DM digestibility (p < 0.001). The pH was highest in CON silage, while lowest in LBR and MIX silages (p < 0.001). The concentrations of ammonia-N, lactate, and acetate were highest in LBR silage (p = 0.008; p < 0.001; p < 0.001). Propionate and butyrate concentrations were highest in CON silage (p = 0.004; p < 0.001). The LAB and yeast counts were higher in CON and LHO silages compare to LBR and MIX silages (p < 0.001). However, the mold did not detect in all treatments. Therefore, this study could conclude that L. brevis 100D8 and Leu. holzapfelii strain 5H4 can improve the digestibility and anti-fungal activity of high moisture rye silage.

The present study investigated effects of microbial additives and silo density on chemical compositions, fermentation indices, and aerobic stability of whole crop rice (WCR) silage. The WCR (“Youngwoo”) was harvested at 49.7% dry matter (DM), and ensiled into 500 kg bale silo with two different compaction pressures at 430 kgf (kilogram-force)/cm2 (LOW) and 760 kgf/cm2 (HIGH) densities. All WCR forage were applied distilled water (CON) or mixed inoculants (Lactobacillus brevis 5M2 and Lactobacillus buchneri 6M1) with 1:1 ratio at 1x105 colony forming unit/g (INO). The concentrations of DM, crude protein, ether extract, crude ash, neutral detergent fiber, and acid detergent fiber of whole crop rice before ensiling were 49.7, 9.59, 2.85, 6.74, 39.7, and 21.9%, respectively. Microbial additives and silo density did not affect the chemical compositions of WCR silage (p>0.05). The INO silages had lower lactate (p<0.001), but higher propionate (p<0.001). The LOW silages had higher lactate (p=0.004). The INO silages had higher yeast count (p<0.001) and aerobic stability (p<0.001). However, microbial counts and aerobic stability were not affected by silo density. Therefore, this study concluded that fermentation quality of WCR silage improved by microbial additives, but no effects by silo density.

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.

The present study was aimed to estimate the effect of ensiling period and bacterial inoculants on chemical compositions and fermentation characteristics on rye silage harvested at delayed stage. Rye (Secale cereale L.) was harvested after 20 days of heading stage (29.4% dry matter, DM). The harvested rye forage was applied with different inoculants following: applications of distilled water (CON), Lactobacillus brevis (LBB), Leuconostoc holzapfelii (LCH), or mixture of LBB and LCH at 1:1 ratio (MIX). Each forage was ensiled into 20 L mini bucket silo (5 kg) for 50 (E50D) and 100 (E100D) days in triplicates. The E50D silages had higher in vitro digestibilities of DM (IVDMD, p<0.001) and neutral detergent fiber (IVNDFD, p=0.013), and lactate (p=0.009), and acetate (p=0.011) than those of E100D, but lower pH, lactic acid bacteria (LAB), and yeast. By inoculant application, LCH had highest IVDMD and IVNDFD (p<0.05), while MIX had highest lactate and lowest pH (p<0.05). The CON and LCH in E50D had highest LAB and yeast (p<0.05), whereas LBB in E100D had lowest (p<0.05). Therefore, this study concluded that LCH application improved the nutrient digesbility (IVDMD and IVNDFD) of lignified rye silage, and longer ensiling period for 100 days enhanced the fermentation characteristics of silage compared to ensiling for 50 days.

This study was conducted to estimate the effect of different cutting lengths on fermentation characteristics and aerobic stability of whole crop rice (WCR) silage. The WCR was harvested at the yellow ripe stage (43.7%, DM), and then cut at 5 (R05), 10 (R10), and 20 cm (R20) of the theoretical length of cut with no cut WCR (R60). Each forage was ensiled into 20 L mini bucket silo (5 kg) for 150 days in quadruplicates. The cutting lengths were not affected the chemical compositions of WCR silage (p > 0.05). The pH (p < 0.001) and concentration of ammonia-N (p = 0.022) in WCR silage were increased linearly with the increase of cutting length. The concentration of lactate had quadratic effect (p = 0.007), which was highest in R20 silage (p < 0.05). The concentration of acetate was increased linearly (p = 0.014), but the concentration of butyrate was decreased linearly (p = 0.033). The lactic acid bacteria count was decreased linearly (p = 0.017), and yeast count had quadratic effect (p = 0.009), which was the highest in R20 silage (p < 0.05). Aerobic stability had strong quadratic effect (p < 0.001), which was the highest in R20 silage (p < 0.05). In conclusion, R60 silage had highest pH by a linear increase of ammonia-N concentration and led to low aerobic stability. While R20 silage had the lowest pH by high lactate concentration and led to high aerobic stability.

This study was to evaluate the feed value of whole crop rice silage (WCRS) and to investigate a suitable ratio of the WCRS and concentrate by an analysis of rumen fermentation. A total of 6 treatments were used according to WCRS: concentrate ratio on in vitro rumen fermentation: T1 (100:0), T2 (60:40), T3 (40:60), T4 (20:80), T5 (10:90), and T6 (0:100). The ruminal pH, total gas emission, ammonia nitrogen, and volatile fatty acid (VFA) were determined as fermentation parameters. Total nutrients digestibility trial was conducted by 4 treatments according to WCRS: concentrate ratio at 40:60 (W40), 20:80 (W20), and 10:90 (W10), respectively. Feed value was analyzed according to AOAC (2019) and nutrient digestibility was calculated based on NRC (2001). The levels of crude protein (CP), crude fat, and neutral detergent fiber of the WCRS were 12.29%, 1.67%, and 59.79%, respectively. It was found to be 51.49% as a result of predicting the total digestible nutrient of WCRS using the NRC (2001) model. In vitro rumen fermentation, T4, T5, and T6 treatments showed a greater gas emission and total VFA concentration compared with other treatments (p<0.05). Acetate and acetate to propionate ratio of T4, T5, and T6 were significantly higher than other treatments (p<0.05). There was a significant difference in the level of propionate and butyrate according to the WCRS: concentrate ratio (p<0.05). The digestibility of dry matter and CP was significantly lower in W40 than in other treatments (p<0.05); however, there was no difference in W20 and W10. In conclusion, the 20:80 (WCRS: concentrate) is beneficial for stabilizing the rumen that does not inhibit rumen fermentation and nutrient digestion. This ratio might have a positive effect on the economics of farms as a valuable feed.

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.

The present study investigated effects of antifungal and carboxylesterase inoculant on rumen fermentation with different rumen pH. Corn silage was treated without inoculant (CON) and with a mixed Lactobacillus brevis 5M2 and L. buchneri 6M1 (MIX). Rumen fluid was collected from two cannulated Hanwoo heifers before morning feeding (high rumen pH at 6.70) and 3 h after feeding (low rumen pH at 6.20). Dried corn silage was incubated in the rumen buffer (rumen fluid + anaerobic culture medium at 1:2 ratio) for 48 h at 39oC. Eight replications for each treatment were used along with two blanks. Both in a high and a low rumen pH, MIX silages presented higher (p<0.05) the immediately degradable fraction, the potentially degradable fraction, total degradable fraction, and total volatile fatty acid (VFA) than those of CON silages. Incubated corn silages in a low rumen pH presented lower (p<0.05) total degradable fraction, ammonia-N, total VFA (p=0.061), and other VFA profiles except acetate and propionate, than those in a high rumen pH. The present study concluded that application of antifungal and carboxylesterase inoculant on corn silage could improve degradation kinetics and fermentation indices in the rumen with high and low pH conditions.

The goal of this study was to evaluate effects of various microbial and organic additives on chemical compositions, fermentation indices, and aerobic stability of barley silage. Youngyang barley harvested at 31.5% dry matter (DM), and ensiled into 20 L bucket silo for 0, 1, 3, 7, 48, and 100 d in quadruplicates with four additives following: sterile destilled water at 1% of fresh forage (CON); Lactobacillus plantarum at 1.2 x 105 cfu/g of fresh forage (CL); Lactobacillus buchneri at rate of 1.2 x 105 cfu/g fresh forage (LB); Fermented Persimmon Extract at 1% of fresh forage (FPE); and Essential Oil at 1% of fresh forage (EO). On 100 d of ensiling, CL and FPE silages had lower (p<0.05) DM than CON silage. Additionally, FPE silage had higher (p<0.05) crude protein than CON silage. All silages with additives, except EO, had higher (p<0.05) neutral detergent fiber (NDF) than CON silage. Silage treated with CL, LB, and FPE had lower in vitro DM digestibility than CON silage, and silages treated with LB and FPE had higher in vitro NDF digestibility (IVNDFD) on 100 d of ensiling. The PFE silage produced the highest (p<0.05) lactate during ensiling period, while LB silage produced the highest (p<0.05) acetate. All inoculated silages had higher (p<0.05) LAB count than control silage. Only CL silage had higher (p<0.05) yeast count than control silage, while the other silages were not differ compared to CON silage. The aerobic stability was higher (p<0.05) in LB and FPE silages than in CON silage. In conclusion, FPE could be an alternative additive to increase IVNDFD, fermentation indices, and aerobic stability of barley silage.

본 연구는 이형발효 유산균을 첨가한 이탈리안 라이그라스 및 총체보리 사일리지의 저장 기간 동안의 변화에 따른 반추위 발효 효과를 규명하고자 수행하였다. 사일리지의 접종 균주는 이형발효 유산균으로 Lactobacillus casei KACC 12413 및 Lactobacillus reuteri KCTC 3594 및 Lactobacillus casei + Lactobacillus reuteri로 하였다. In vitro 실험의 pH 변화는 LCR 처리구가 높은 경향이었으며, 작물 간의 차이는 WCB사일리지 보다 IRG 사일리지의 pH가 높았다. 총 가스 생산량은 대조구에 비해 유산균 첨가구가 낮았고, WCB 보다 IRG 사일리지가 높은 수준이었다. 암모니아태 질소 농도는 유산균 첨가구 보다 대조구가 높은 경향이었고, 30일 보다 60일 사일리지가 낮았다. 총 휘발성지방산 생성량은 30일 및 60일 WCB 사일리지의 LC 처리구가 높았다. In situ 실험에서 건물 소화율은 48시간 발효 했을 때 LCR 처리구가 높았고 대조구가 가장 낮았다. 작물 간에 차이는 WCB에 비해 IRG가 8∼10% 더 높았다. NDF 소화율은 LC 처리구 및 LCR 처리구가 높은 경향이었고(p<0.05), ADF 소화율은 LC 처리구 및 LCR처리구가 높았다. 본 연구의 결과로 미루어 보아 propionate를 생성하는 이형 발효 유산균이 접종된 사일리지는 반추위 내 발효조건을 개선하고 가축의 사일리지 소화율을 증진 시킬 것으로 보이며, 가축의 생산성 및 사료가치가 향상 될 것으로 여겨진다.

The purpose of this study was to analyze the effectiveness of different storage periods of lactic acid bacteria (LAB)-fermented low moisture fresh rice straw silage. The low moisture fresh rice straw sample was inculcated with LAB and stored for different storage periods such as 45, 90, and 365 days, respectively. The low moisture fresh rice straw (LMFRS) silage inoculated with LAB exhibited reduction in pH throughout the fermentation as compared with the control (P<0.05). The lactic acid content was increased at the late fermentation period (90 and 365 days, respectively) in LAB inoculated LMFRS silage as compared with the control (P<0.05). In contrast, the acetic acid and butyric acid concentrations were slightly reduced in the LAB inoculated LMFRS silage sample at 90 and 365 days fermentation, respectively. Meanwhile, the non-inoculated LMFRS silage showed higher amounts of acetic acid and butyric acid at an extended fermentation with low bacterial population as compared with the LAB inoculated LMFRS silage. However, lactic acid concentration was slightly high in the non-inoculated LMFRS silage at early 45 days fermentation. Additionally, the nutrient profile such as crude protein (CP), acid detergent fibre (ADF), neutral detergent fibre (NDF), and total digestibility nutrients (TDN) were not significantly different in control and LAB inculcated samples during all fermentation. Though, the microbial population was greater in the LAB inoculated LMFRS silage as compared with the control. However, the massive population was noted in the LAB inoculated LMFRS silage during all fermentation. It indicates that the inoculated LAB is the main reason for increasing fermentation quality in the sample through pH reduction by organic acids production. Overall results suggest that the LAB inoculums are the effective strain that could be a suitable for LMFRS silage fermentation at prolonged days.

The effects of lactic acid bacteria (LAB) mixtures on low moisture Italian ryegrass (IRG) silage fermentation was evaluated in field conditions. The experiment was categorized into two groups: Un-inoculated (Control) and Inoculated with LAB mixture for four storage periods (45, 90, 180, and 365 days, respectively). Silage inoculated with the LAB mixture had the lowest pH with highest lactic acid production than the control from beginning at 45-365 days at all moistures. Higher LAB counts were observed in inoculated silages than the control silages at whole experimental periods. It is a key reason for the rapid acidification and higher lactic acid production in silages during the storage periods. Overall results suggest that an adding of LAB mixture had positive effects on the increasing aerobic stability of silage and preserved its quality for an extended duration.