Effect of supplementing herbal feed additives on the fermentation pattern and In vitro methane production by total mixed rations with different roughage to concentrate ratios

Simple SummaryRoughage type and particle size may play an important role in rumen volatile fatty acid levels and consequently affect fatty acid profile in meat and quality. Two roughage sources and two particles were used in this trial for growing lambs to study the effect in volatile fatty acids levels and fatty acids profile of longissimus dorsi (LD) fat. The results of this trial supported the hypothesis by confirming the effect of roughage type and particle size on meat quality and human health.The fatty acid composition for the longissimus dorsi (LD) fat of carcass sheep is a crucial factor impacting meat quality. We performed a 90-day feeding trial of 25 Naemi lambs to investigate the effects of roughage sources (alfalfa or wheat straw) of two sizes (regular and 1 cm chopped) when fed with pelleted total mixed ration (TMR) on the growth performance, fermentation patterns, and fatty acid (FA) composition of longissimus dorsi (LD) fat. Lambs were randomly assigned to individual pens with five treatment diets, as follows: C, control group with TMR; T1, TMR and regular alfalfa hay; T2, TMR and alfalfa hay chopped to 1 cm; T3, TMR and regular wheat straw; and T4, TMR and wheat straw chopped to 1 cm. Four lambs were randomly selected from each treatment (20 total) and sacrificed. LD fat of the carcass was extracted and analyzed for FA using a gas chromatography-mass spectrometry. Significantly increased feed intake was found in T1 and T2. The FA composition of LD fat in T2 had higher unsaturated fatty acid (UFA), omega-6 (n6), and omega-3 (n3) FA content. Conjugated linoleic acid (CLA) and α-linoleic acid were highest in lambs fed T1 and T2. Feeding different types of roughage, especially alfalfa hay, either regular or chopped, with total pelleted mixed ration is crucial to improving feed intake and body weight gain, as it positively enhances the rumen microbial fermentation process by controlling rumen pH. The FA profiles of meat from lambs fed TMR with regular or 1 cm particle size alfalfa hay (T1 and T2) are recommended for human consumption as a source of healthy FAs.

Total mixed ration (TMR) containing roughage and concentrate mixture in a 65: 35 ratio on dry matter (DM) basis was supplemented with herbal feed additives [(HFAs); Acacia catechu (Katha), Areca catechu (Supari), and Acacia nilotica (Babul)] at 0-4% on DM basis to assess their impact on fermentation pattern and methane production by using in vitro gas production technique. Areca catechu had the highest (P < 0.01) concentration of condensed tannins (CTs), saponins, and vitamin C as compared to other HFAs. The net gas production (NGP), digestibility of NDF and true OM, and ME availability, partitioning factor (PF), volatile fatty acids (VFAs), and microbial biomass production were higher (P < 0.01) at 24 h as compared to t-half incubation, irrespective of type and level of HFAs supplemented. Acacia nilotica had an edge over Acacia catechu with respect to digestibility of nutrients and ME availability. VFAs production and efficiency of rumen fermentation was the highest from the Areca catechu-supplemented TMR. Irrespective of type of HFAs and incubation period, the digestibility of NDF and that of true OM were highest (P < 0.01) at the 1% level of supplementation, but depressed thereafter. Amongst HFAs-supplemented groups, VFA production and fermentation efficiency were highest at the 2% level. The results conclusively revealed that supplementing Areca catechu and Acacia nilotica at 2% of TMR (DM basis) inhibited the methane production, without affecting the fermentation pattern.

This study was taken up to assess the impact of supplementing herbal feed additives [HFAs; fruit of Myristica fragrans (Jayphall), seeds of Anethum sowa (Suva), fruit of Apium graveolens (Ajmo), fruit of Cuminum cyminum (Jeera), bark of Cinnamonum zeylanicum (Dalchini), or whole plant of Eclipta alba (Bhangro)] containing essential oils as active component on the nutrient utilization and methane production using wheat straw-based total mixed ration (TMR) as a substrate by in vitro gas production technique. The essential oil content was the highest (P < 0.01) in M. fragrans followed by E. alba and A. sowa. In addition to essential oils, these HFAs also contained saponins, tannins, and antioxidants. The HFAs were supplemented at 1-3% of substrate dry matter (DM). The data were analyzed by 6 × 4 factorial design. Irrespective of level of HFA, the net gas production (NGP) and metabolizable energy (ME) availability was the highest (P < 0.01) in TMR supplemented with C. zeylanicum comparable with E. alba, but higher than TMR supplemented with other HFAs. Supplementation of TMR with different HFAs did not affect the digestibility of neutral detergent fiber (NDF) and true organic matter (TOM) and partitioning factor (PF). The total volatile fatty acids (VFAs), acetate, propionate (P < 0.01), and butyrate (P < 0.05) production was the highest in TMR supplemented with A. sowa, and the lowest was observed in TMR supplemented with C. cyminum. The isobutyrate and valerate production was also the highest (P < 0.01) in diet supplemented with A. sowa, but isovalerate production was the highest (P < 0.01) in diet supplemented with C. zeylanicum. The A:P ratio was the best in TMR supplemented with A. sowa. The efficiency of rumen fermentation was the highest, and efficiency of conversion of hexose to methane was the lowest in diet supplemented with A. sowa as compared to all other supplements. The in vitro methane production expressed as either percent of NGP, ml/100mg DM of substrate/24h, or as ml/100mg of digestible OM/24h was the lowest in TMR supplemented with A. sowa. The ammonia nitrogen production from TMR supplemented with M. fragrans and A. sowa was comparable, but significantly (P < 0.01) lower than TMR supplemented with other HFAs. Irrespective of the nature of HFA, the NGP and ME availability were significantly (P < 0.01) higher in TMR supplemented with HFAs at all levels as compared to un-supplemented TMR. As compared to control, the digestibility of NDF and that of TOM was depressed slightly in all the HFA-supplemented TMRs. The supplementation of HFAs at 2% of substrate DM improved (P < 0.01) the production of total VFAs, acetate, and propionate, and that of isovalerate in comparison to the un-supplemented TMR. The acetate to propionate ratio increased (P < 0.01) with the increase in the level of supplementation of HFAs containing essential oils. The methane and ammonia productions were depressed significantly when TMR was supplemented at 2% level of HFAs as compared to control TMR. It was concluded that supplementation of TMR with A. sowa at 2% of substrate was fermented better as indicated by the production of total and individual VFA, methane, and ammonia as compared to TMR supplemented with other HFA or un-supplemented TMR.

This study was taken up to assess the effect ofdry extract of six herbal feed additives [HFAs; Kulthi (Dohichos biflorus), patha (Cissampelospareria), aritha (Sapindustrifoliatus), methi (Trigonella foenum graecum), shikakai (Acacia concina) and shatavari (Asparagus racemosus)] rich in saponins supplemented at 1–4% of total mixed ration (TMR) on DM basis on the in vitro fermentation pattern and methane emission. A TMR with roughage to concentrate (R:C) ratio of 70:30 on DM basis was formulated. The data was analyzed by 2 x 6 x 5 factorial design. The net gas production (NGP), digestibility of NDF and OM, ME availability, total and individual volatile fatty acids (VFAs) were higher (P<0.01) at 24hincubation in comparison to that observed at t'A The NGP and ME in the TMR supplemented with shatavari was higher (P<0.01) than all other HFAs supplemented groups and increased (P<0.01) with the increase in the level of HFAs. The total and individual VFAs were the highest (P<0.01)when the diet was supplemented with HFAs at 3% levels. The methane production was lowest(P<0.01) and the fermentation efficiency was the highest in TMR supplemented with shatavari. The methane emission was depressed at all levels of HFAs as compared to control group.The methane emission at t-half was observed to be the lowest in the TMR supplemented with patha, kulthi or shatavari at 2% on DM basis.It was concluded that best response in above parameters could be ascertained in TMR supplemented with shatavari @ 2–3% on DM basis.

Simple SummaryMethane is a by-product of the microbial fermentation process from a group of archaea known as methanogens. Ruminants harbor methanogens as a component of their normal gastrointestinal microbiota. In this study, we assessed the fecal microbiome of organic dairy cows across different time points receiving a mixed diet of pasture and TMR or TMR only. Our aim was to compare the archaea and bacterial fecal microbial structure, diversity and their interactions across diets. Cluster analysis based on the cows’ fecal microbial community identified four clusters. We found little difference in the relative abundance of methanogens across clusters. However, there was evidence of differences in diversity between pasture associated communities and those associated with TMR only. The cluster associated with cows receiving high-quality pasture and TMR, had higher diversity and a less robust co-occurrence network than those in TMR only or lower-quality pasture communities. The overall good pasture and TMR quality, combined with the organic allowance for feeding high levels of TMR, might have contributed to the lack of differences in the fecal archaeal community. The development of novel strategies that are independent of pasture management could have a greater impact in helping to reduce enteric CH4 emissions on organic dairies.Currently, little is known regarding fecal microbial populations and their associations with methanogenic archaea in pasture-based dairy cattle. In this study, we assessed the fecal microbiome of organic dairy cows across different time points receiving a mixed diet of pasture and total mixed ration (TMR) or TMR only. We hypothesized that the fecal methanogenic community, as well as co-occurrence patterns with bacteria, change across diets. To test these hypotheses, we analyzed TMR and pasture samples, as well as the V3-V4 region of 16S rRNA of fecal samples collected over the course of a one-year study period from 209 cows located on an organic dairy in Northwest Washington. The inherent variability in pasture quality, quantity, availability, and animal preference can lead to diverse dietary intakes. Therefore, we conducted a k-means clustering analysis to identify samples from cows that were associated with either a pasture-based diet or a solely TMR diet. A total of 4 clusters were identified. Clusters 1 and 3 were mainly associated with samples primarily collected from cows with access to pasture of varying quality and TMR, cluster 2 was formed by samples from cows receiving only TMR, and cluster 4 was a mix of samples from cows receiving high-quality pasture and TMR or TMR only. Interestingly, we found little difference in the relative abundance of methanogens between the community clusters. There was evidence of differences in diversity between pasture associated bacterial communities and those associated with TMR. Cluster 4 had higher diversity and a less robust co-occurrence network based on Spearman correlations than communities representing TMR only or lower-quality pasture samples. These findings indicate that varied bacterial communities are correlated with the metabolic characteristics of different diets. The overall good pasture and TMR quality in this study, combined with the organic allowance for feeding high levels of TMR even during the grazing season, might have contributed to the lack of differences in the fecal archaeal community from samples associated with a mixed pasture and TMR diet, and a TMR only diet. Mitigation strategies to decrease methane emissions such as increasing concentrate to forage ratio, decreasing pasture maturity and adopting grazing systems targeting high quality pasture have been shown to be efficient for pasture-based systems. However, the allowance for organic dairy producers to provide up to an average of 70% of a ruminant’s dry matter demand from dry matter fed (e.g., TMR), suggests that reducing enteric methane emissions may require the development of novel dietary strategies independent of pasture management.

The objective of this study was to examine the conservation process and feed value of total mixed ration (TMR) silages. In exp. 1, we evaluated the fermentation pattern and aerobic stability of TMR silages containing different protein and lipid supplementations. In exp. 2, we compared the performance of finishing beef heifers fed those TMR silages. In both experiments, treatments were as follows: ensiled TMR with urea (U); ensiled TMR without a protein supplement at ensiling, but soybean meal supplemented at feeding to balance diet crude protein (CP) in exp. 2 (SMnf; where the acronym nf indicates nonfermented); ensiled TMR with soybean meal (SM); and ensiled TMR with rolled soybean grain (SG). Thirty-two Nellore heifers (313 ± 8.8 kg shrunk body weight [SBW]) were blocked by initial SBW, housed in individual pens, and enrolled in exp. 2 for 82 d. In exp. 1, treatment without a protein supplement (SMnf) had a lower content of CP, soluble CP, NH3-N, pH, and Clostridium count compared with U (P ≤ 0.03). Lactic acid concentrations tended to be reduced for SMnf compared with U (P = 0.09). Ethanol concentration was reduced in SG compared with SM (P < 0.01). 1,2-Propanediol concentration was increased in SMnf compared with U (P < 0.01), reduced in SM compared with SMnf (P = 0.02), and increased in SG compared with SM (P = 0.02). Dry matter (DM) loss during fermentation was low and similar among treatments (~3.7%). All silages remained stable during 10 d of aerobic exposure after feed out. Considering fermentation traits, such as pH (≤4.72), NH3-N (<10% of N, except for U treatment), butyric acid (<0.05 % DM), and DM losses (<3.70% DM), all silages can be considered well conserved. In exp. 2, diets were isonitrogenous because soybean meal was added to SMnf before feeding. Compared with SM, cattle fed SG made more meals per day (P = 0.04) and tended to have a decreased intermeal interval (P = 0.09). DM intake, average daily gain, final SBW, hot carcass weight, Biceps femoris fat thickness, and serum levels of triglycerides and cholesterol were increased for SG compared with SM (P ≤ 0.05). In brief, TMR silages exhibited an adequate fermentation pattern and high aerobic stability. The supplementation of true protein did not improve animal performance, whereas the addition of soybean grain as a lipid source improved the performance of finishing cattle fed TMR silages.

The objectives of this study were to evaluate lauric acid (LA) as a practical ruminal protozoa-suppressing agent and assess effects of protozoal suppression on fermentation patterns and milk production in dairy cows. In a pilot study, 6 lactating Holstein cows fitted with ruminal cannulae were used in a randomized complete-block design trial. Cows were fed a basal total mixed ration (TMR) containing (DM basis) 15% alfalfa silage, 40% corn silage, 30% rolled high moisture shelled corn, and 14% solvent soybean meal, and assigned to 1 of 3 treatments: 1) control, 2) 160 g/d of LA, or 3) 222 g/d of sodium laurate, which is equimolar to 160 g/d of LA, all given as a single dose into the rumen via cannulae before feeding. Both agents showed high antiprotozoal activity when pulse dosed at these amounts via ruminal cannulae, reducing protozoa by 90% (P<0.01) within 2 d of treatment. Lauric acid reduced ruminal ammonia concentration by 60% (P<0.01) without altering DMI. Both agents reduced ruminal total free AA concentration (P<0.01) and LA did not affect ruminal pH or total VFA concentration. In a large follow-up feeding trial, 52 Holstein cows (8 with ruminal cannulae) were used in a randomized complete-block design trial. Cows were assigned to 1 of 4 diets and fed only that diet throughout the study. The TMR contained (DM basis) 29% alfalfa silage, 36% corn silage, 14% rolled high moisture shelled corn, and 8% solvent soybean meal. The 4 experimental diets were similar, except part of the finely ground dry corn was replaced with LA in stepwise increments from 0 to 0.97% of dietary DM, which provided (as consumed) 0, 83, 164, and 243 g/d of LA. Adding these amounts of LA to the TMR did not affect DMI, ruminal pH, or other ruminal traits, and milk production. However, LA consumed at 164 and 243 g/d in the TMR reduced the protozoal population by only 25% and 30% (P=0.05), respectively, showing that these levels, when added to the TMR, were not sufficient to achieve a concentration within the rumen that promoted the antiprotozoal effect of LA.

Two, 8-week experiments, each using 30 lactating Holstein cows, were conducted to examine performance of animals offered combinations of total mixed ration (TMR) and high-quality pasture. Experiment 1 was initiated in mid October 2004 and Experiment 2 was initiated in late March 2005. Cows were assigned to either a 100% TMR diet (100:00, no access to pasture) or one of the following three formulated partial mixed rations (PMR) targeted at (1) 85% TMR and 15% pasture, (2) 70% TMR and 30% pasture and (3) 55% TMR and 45% pasture. Based on actual TMR and pasture intake, the dietary TMR and pasture proportions of the three PMR in Experiment 1 were 79% TMR and 21% pasture (79:21), 68% TMR and 32% pasture (68:32), and 59% TMR and 41% pasture (59:41), respectively. Corresponding proportions in Experiment 2 were 89% TMR and 11% pasture (89:11), 79% TMR and 21% pasture (79:21) and 65% TMR and 35% pasture (65:35), respectively. Reducing the proportion of TMR in the diets increased pasture consumption of cows on all PMR, but reduced total dry matter intake compared with cows on 100:00. An increase in forage from pasture increased the concentration of conjugated linoleic acids and decreased the concentration of saturated fatty acids in milk. Although milk and milk protein yields from cows grazing spring pastures (Experiment 2) increased with increasing intakes of TMR, a partial mixed ration that was composed of 41% pasture grazed in the fall (Experiment 1) resulted in a similar overall lactation performance with increased feed efficiency compared to an all-TMR ration.

This study was undertaken to screen the tree leaves (TLs) for bio-active compounds and assess their impact on the substrate degradation and methane production from total mixed ration (TMR) by in-vitro gas production technique. The sundried, finely ground leaves of Mangifera indica (MI; Mango), Cassia fistula (CF; Amaltas/Amulthus) and Acacia nilotica (AN; Babul) were supplemented individually and in combinations i.e. MI+CF, MI+AN, CF+AN and MI+CF+AN to TMR at 1% level on DM basis. Results revealed that the total phenols, condensed tannins, hydrolysable tannins, DPPH and vitamin C content were the highest (P<0.01) in Mangifera leaves. The saponins content was the highest (P<0.01) in Acacia leaves. Irrespective of incubation period, the net gas production (NGP) and ME were the highest (P<0.01) in TMR supplemented with combination of MI+AN. The digestibility of NDF and true OM was highest (P<0.01) in TMR supplemented with Cassia leaves, which was comparable with that supplemented with Mangifera, MI+AN and unsupplemented TMR. The highest (P<0.01) concentration of total and individual VFAs in TLs supplemented groups was observed in TMR supplemented with Mangifera leaves, followed by that supplemented with Casia leaves. The in vitro methane production was reduced significantly (P<0.01) in all the TLs supplemented groups except that supplemented with Mangifera leaves which was higher than unsupplemented TMR. The best fermentation efficiency was observed in TMR supplemented with MI+CF+AN followed by that supplemented with Casia and Mangifera leaves. The microbial biomass synthesized in TMR supplemented with Mangifera leaves was higher (P<0.01) than that of Casia supplemented and unsupplemented TMR. Irrespective of TLs supplementation, all the above parameters were higher (P<0.01) at 24h incubation as compared to that at t-half, except that the efficiency of conversion of fermented hexose to methane was higher (p<0.01) at t-half as compared to 24h incubation. It was concluded that Mangifera leaves is a rich source of bioactive compounds. Supplementation of Mangifera and Casia leaves resulted in the highest digestibility of nutrients, VFAs production, efficiency of rumen fermentation and microbial biomass synthesis; and decrement in efficiency of conversion of fermented hexose to methane from TMR.

An in vitro study was conducted to assess the effect of using rice gluten meal as a substitute of soybean meal (SBM) in ruminant diet. The SBM comprising 12.5% whole diet [total mixed ration (TMR)] was substituted by rice gluten meal (RGM) on w/w basis at 0, 25, 50, 75 and 100% level. In vitro dry matter disappearance (IVDMD), organic matter disappearance (IVOMD), net gas production, methane production and fermentation efficiency of TMRs was assessed. The chemical composition of TMRs revealed decrease in total ash, acid detergent fiber and total carbohydrate contents and increase in organic matter, crude protein, ether extract, neutral detergent fiber, acid detergent lignin, hemicellulose, neutral detergent insoluble crude protein and acid detergent insoluble crude protein contents with increase in RGM in TMRs. No significant difference in net gas production, IVDMD and IVOMD was observed between the TMR treatments having graded levels of RGM and control. The NH3-N in TMRs declined with increase in RGM and was highest (P<0.05) in control. The relative proportion of propionic acid and isovaleric acid was highest in TMR 3 (50% RGM replacing SBM); whereas relative proportion of butyric acid was lowest in TMR 3. CH4 production showed decline with increase in RGM in TMRs beyond 25% level replacing SBM. The H-recovery was lowest in control TMR. The hydrogen consumed via CH4 as compared to the hydrogen consumed via VFA was lowest in TMR 3. The fermentation efficiency was highest in TMR 3. Thus, RGM could replace up to 100% of soybean meal in TMR of ruminants without any adverse effect on nutrient digestibility which is equivalent to 12.5% of whole dietary dry matter.

The aim of the present study was to evaluate the effect of feeding green fodder, rice straw and concentrate-based total mixed rations (TMR) on dry matter (DM) intake (DMI), nutrient utilization, rumen fermentation patterns and body weight (BW) gain (BWG) in mithun (Bos frontalis) calves. In a randomized block design, male mithun calves (n = 18, 8-10 months of age, 121 +/- 2 kg BW) were randomly divided into three experimental equal groups (six animals in each group) and fed isonitrogenous TMRs ad libitum for 120 days. The TMR(1) contained 30% Napier grass and 30% rice straw, TMR(2) contained 60% rice straw and TMR(3) contained 30% tree leaves (Lagerstroemia speciosa) and 30% rice straw (DM basis). All the TMRs contained 40% concentrate mixture (DM basis). The results indicated that the BWG, DMI and feed conversion efficiency were significantly (p < 0.01) increased with the inclusion of green fodder in TMRs. The apparent digestibility of DM, crude protein, ether extract, crude fibre and nitrogen free extract were also improved significantly (p < 0.01) with the inclusion of green fodder in TMRs. The higher concentration of total nitrogen and total volatile fatty acid in rumen liquor, but low ruminal pH were evident in animals fed green fodder supplemented TMRs. An increased (p < 0.01) molar proportion of acetic acid was evident in animals fed rice straw-based TMR. In contrast, the molar proportion of propionic and butyric acids were significantly (p < 0.01) higher in animals fed green fodder supplemented TMRs. On the basis of higher DMI and higher daily BWG, it is concluded that Napier grass and L. speciosa tree leaves may be incorporated upto 30% (DM basis) in TMR of growing mithuns for feeding in complete confinement system.

Effect of feeding forest foliages, rice straw and concentrate-based total mixed ration on nutrient utilization and growth in mithun ( Bos frontalis)

Effects of altering total mixed ration conservation method when feeding dry-rolled versus steam-flaked hulled rice on lactation and digestion in dairy cows

Accuracy and homogeneity of total mixed rations processed through trailer mixer or self-propelled mixer, and effects on the yields of high-yielding dairy cows

본 연구의 목적은 TMR 제조 시 물을 첨가하여 수분 함량을 높여줄 경우 수분 함량(15, 35, 50%)과 발효제(Lactobacillus plantarum) 첨가유무에 따라 저장방법(자연노출과 밀봉보관) 경과시간별(자연노출 : 0, 6, 12, 24, 48, 72시간, 밀봉보관 : 0, 2, 6, 10, 14, 18일) 사료가치 변화를 조사할 목적으로 실시하였으며 그 결과는 다음과 같다. 자연노출 상태로 저장한 경우, 수분 함량이 적을수록 외관상태가 양호하였으며 수분 함량이 높을수록 곰팡이가 생기는 시기가 빨라짐을 알 수 있었다. 시료의 냄새도 수분함량별, 발효제 첨가유무별로 달랐는데 물만 첨가하여 수분 함량을 35%와 50%로 맞춘 시료는 자연노출 24시간째에 시큼한 냄새가 나기 시작하였으나 같은 수분 함량의 시료에 발효제를 첨가한 시료에서는 발효에 의한 냄새로 시큼한 냄새가 나지 않았고 오히려 발효로 인해 향긋한 냄새가 났다. 발효제를 첨가하지 않은 시료에서는 자연노출 후 6시간이 경과하면서 지속적으로 내부온도가 상승하는 경향을 나타내고 있으며, 발효제를 첨가한 수분 함량 50% 시료에서는 자연노출 6시간부터 지속적으로 온도가 상승하는 경향을 나타낸 반면, 발효제를 첨가한 수분 35% 시료는 자연노출 48시간부터 온도가 급격히 상승함을 보이고 있다. 발효제를 첨가한 시료를 자연노출 시 수분 함량에 따른 경과시간별 pH 변화를 보면, 수분 15%에서는 경과시간에 따른 변화가 적었으나 수분 35%와 50% 시료는 각각 12시간째와 24시간째부터 pH가 급격하게 증가 하였다. 자연노출 시 같은 수분 함량의 TMR 시료에서 발효제를 첨가한 것은 6시간까지는 암모니아 발생이 낮아지다가 6시간 이후부터 암모니아 농도가 증가하는 경향을 나타낸 반면, 발효제를 첨가하지 않은 것은 12시간 이후에 암모니아 농도가 증가하는 경향을 나타내었다. 자연노출 시 영양소 함량 변화는 경과시간이 진행됨에 따라 수분의 증발로 인해 영양소 함량이 증가하는 경향을 나타내었으며, 밀봉보관 시에도 같은 경향을 나타내었다. 이상과 같은 결과를 종합해 보면, TMR 제조 시 수분함량을 조절하기 위하여 물을 첨가시킬 때 물만 첨가하는 것보다는 물에 발효제(Lactobacillus plantarum)를 섞어 주는 것이 경과시간에 따른 TMR의 사료가치 저하를 방지할 수가 있을 것으로 사료되었다. This study was carried out to investigate effects of increasing moisture content with or without supplementing inoculant (Lactobacillus plantarum) in TMR (total mixed ration) on its feed value. In case of exposing TMR to air, the lower the moisture level of TMR was, the less its apparent condition was changed. The time of spreading of molds tended to be faster in TMR with the higher moisture level. And also the odor was influenced by moisture content and inoculant supplement that is, sour odor was smelled from 24 hour after exposing TMR containing 35% and 50% moisture to air, but TMR supplemented with inoculant had sweet odor. The inner temperature of TMR containing 35% and 50% moisture without inoculant tended to increase continually after the lapse of 6 hours when the TMR was exposed to air. The inner temperature of TMR containing 35% moisture with inoculant tended to increase dramatically after the lapse of 48 hours when exposed to air, but that of TMR containing 50% moisture with inoculant tended to increase after 6 hours. The pH of TMR containing 15% was consistent regardless of exposing time to air, but that of TMR containing 35% and 50% moisture considerably increased after 12 and 24 hours, respectively. The concentration of <TEX>$NH_3-N$</TEX> of TMR supplemented with inoculant was increased from 6 hours after exposure to air, while that or TMR without inoculant increased from 12 hours. Nutrient content or TMR tended to be increased with the increase of exposing time to air and storage time under sealed condition.