• Korean Journal of Organic Agriculture
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• v.21 no.3
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• pp.437-450
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• 2013

Miscanthus sacchariflorus var. No. 1 has been newly developed in Korea. This study was conducted to assess the feed value of M. sacchariflorus var. No. 1 at different growth and harvesting time. Total 3 different miscanthus - 1y4m (first shoot and harvested at 4 month), 2y4m (second shoot and harvested at 4 month) and 2y8m (second shoot and harvested at 8 month). Two experiments were carried out, In vitro rumen simulated fermentation and In situ dry matter digestibility (DMD). Ruminal pH at in vitro fermentation were higher in M. sacchariflorus var. No. 1 treatments compared to the rice straw (RS). In volatile fatty acid production, 1y4m resulted in higher acetate production than the other M. sacchariflorus var. No. 1 at higher maturity stages. Significant differences among treatments were observed in propionate and total volatile fatty acid (VFA) productions at 9, 24 and 48 h of incubation times. Higher ammonia nitrogen productions were found as increased maturity of M. sacchariflorus var. No. 1. At In situ experiment, high DMD was detected in the order of RS (60.51%) > 1y4m (57.65%) > 2y4m (57.63%) > 2y8m (46.28%). The results from this study indicate that young and early harvested M. sacchariflorus var. No. 1 are able to improve its nutrient values in the ruminant animal.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.5
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• pp.687-694
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• 2002

An in vitro study was conducted to examine the effect of addition level of concentrate on fermentation characteristics and long-chain unsaturated fatty acids composition, especially conjugated linoleic acid (CLA) and trans-octadecenoic acid (t-FA) by mixed ruminal bacteria when incubated with linseed or rapeseed. Four levels (0.83, 1.25, 1.67 and 2.08%, w/v) of concentrate and ground oilseeds (linseed or rapeseed; 0.83%, w/v) were added to mixed solution of strained rumen fluid with artificial saliva (1:1, v/v) in the glass jar with a glass lid equipped with stirrer, and was incubated anaerobically for 24 h at $39^{\circ}C$. Addition level of concentrate slightly reflect on pH and ammonia concentration of the culture solution at the various incubation times when incubated with both linseed and rapeseed. Total VFA concentration slightly increased with incubation times and concentrate levels for incubations with oilseeds. While CLA composition had a clearly increasing trend with incubation time when incubated with linseed, percent CLA was relatively stable when incubated with rapeseed. Percent CLA, however, had a clearly decreasing trend with concentrate level throughout incubation times with significances at 3 h incubations when incubated with linseed (p<0.038) and rapeseed (p<0.0009). The differences in compositions of t-FA were relatively small among concentrate levels for both incubations with linseed and rapeseed. The ratios of t-FA to CLA were lower for linseed with increased proportion of CLA than for rapeseed.

• Journal of Animal Science and Technology
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• v.65 no.2
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• pp.387-400
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• 2023

Ruminal protozoa, especially entodiniomorphs, engulf other members of the rumen microbiome in large numbers; and they release oligopeptides and amino acids, which can be fermented to ammonia and volatile fatty acids (VFAs) by amino acid-fermenting bacteria (AAFB). Studies using defaunated (protozoa-free) sheep have demonstrated that ruminal protozoa considerably increase intraruminal nitrogen recycling but decrease nitrogen utilization efficiency in ruminants. However, direct interactions between ruminal protozoa and AAFB have not been demonstrated because of their inability to establish axenic cultures of any ruminal protozoan. Thus, this study was performed to evaluate the interaction between Entodinium caudatum, which is the most predominant rumen ciliate species, and an AAFB consortium in terms of feed degradation and ammonia production along with the microbial population shift of select bacterial species (Prevotella ruminicola, Clostridium aminophilum, and Peptostreptococcus anaerobius). From an Ent. caudatum culture that had been maintained by daily feeding and transfers every 3 or 4 days, the bacteria and methanogens loosely associated with Ent. caudatum cells were removed by filtration and washing. An AAFB consortium was established by repeated transfers and enrichment with casamino acids as the sole substrate. The cultures of Ent. caudatum alone (Ec) and AAFB alone (AAFB) and the co-culture of Ent. caudatum and AAFB (Ec + AAFB) were set up in three replicates and incubated at 39℃ for 72 h. The digestibility of dry matter (DM) and fiber (NDF), VFA profiles, ammonia concentrations, pH, and microscopic counts of Ent. caudatum were compared among the three cultures. The co-culture of AAFB and Ent. caudatum enhanced DM degradation, VFA production, and Ent. caudatum cell counts; conversely, it decreased acetate: propionate ratio although the total bacterial abundance was similar between Ec and the Ec + AAFB co-culture after 24 h incubation. The ammonia production and relative abundance of C. aminophilum and P. anaerobius did not differ between AAFB alone and the Ec + AAFB co-culture. Our results indicate that Ent. caudatum and AAFB could have a mutualistic interaction that benefited each other, but their interactions were complex and might not increase ammoniagenesis. Further research should examine how such interactions affect the population dynamics of AAFB.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.1
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• pp.82-89
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• 2009

Two in vitro experiments were conducted to examine the effects of propionate precursors in the dicarboxylic acid pathway on ruminal fermentatation characteristics, $CH_4$ production and degradation of feed by rumen microbes. Fumarate or malate as sodium salts (Exp. 1) or acid type (Exp. 2) were added to the culture solution (150 ml, 50% strained rumen fluid and 50% artificial saliva) to achieve final concentrations of 0, 8, 16 and 24 mM, and incubated anaerobically for 0, 1, 3, 6, 9 and 12 h at $39^{\circ}C$. For both experiments, two grams of feed consisting of 70% concentrate and 30% ground alfalfa (DM basis) were prepared in a nylon bag, and were placed in a bottle containing the culture solution. Addition of fumarate or malate in both sodium salt and acid form increased (p<0.0001) pH of culture solution at 3, 6, 9 and 12 h incubations. The pH (p<0.0001) and total volatile fatty acids (VFA, p<0.05) were enhanced by these precursors as sodium salt at 3, 6 and 9 h incubations, and pH (p<0.001) and total VFA (p<0.01) from fumarate or malate in acid form were enhanced at a late stage of fermentation (9 h and 12 h) as the addition level increased. pH was higher (p<0.001) for fumarate than for malate as sodium salt at 3 h and 6 h incubations. Propionate ($C_3$) proportion was increased (p<0.0001) but those of $C_2$ (p<0.05) and $C_4$ (p<0.01 - p<0.001) were reduced by the addition of sodium salt precursors from 3 h to 12 incubation times while both precursors in acid form enhanced (p<0.011 - p<0.0001) proportion of $C_3$ from 6h but reduced (p<0.018 - p<0.0005) $C_4$ proportion at incubation times of 1, 3, 9 and 12 h. Proportion of $C_3$ was increased (p<0.05 - p<0.0001) at all incubation times by both precursors as sodium salt while that of $C_3$ was increased (p<0.001) from 6h but $C_4$ proportion was decreased by both precursors in acid form as the addition level increased. Proportion of $C_3$ was higher (p<0.01 - p<0.001) for fumarate than malate as sodium salt from 6 h incubation but was higher for malate than fumarate in acid form at 9 h (p<0.05) and 12 h (p<0.01) incubation times. Increased levels (16 and 24 mM) of fumarate or malate as sodium salt (p<0.017) and both precursors in acid form (p<0.028) increased the total gas production, but no differences were found between precursors in both chemical types. Propionate precursors in both chemical types clearly reduced (p<0.0001 - p<0.0002) $CH_4$ production, and the reduction (p<0.001 - p<0.0001) was dose dependent as the addition level of precursors increased. The $CH_4$ generated was smaller (p<0.01 - p<0.0001) for fumarate than for malate in both chemical types. Addition of fumarate or malate as sodium type reduced (p<0.004) dry matter degradation while both precursors in both chemical types slightly increased neutral detergent fiber degradability of feed in the nylon bag.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.4
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• pp.500-508
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• 2016

This study was conducted to investigate the effects of Momordica charantia saponin (MCS) on ruminal fermentation of maize stover and abundance of selected microbial populations in vitro. Five levels of MCS supplements (0, 0.01, 0.06, 0.30, 0.60 mg/mL) were tested. The pH, $NH_3-N$, and volatile fatty acid were measured at 6, 24, 48 h of in vitro mixed incubation fluids, whilst the selected microbial populations were determined at 6 and 24 h. The high dose of MCS increased the initial fractional rate of degradation at t-value = 0 ($FRD_0$) and the fractional rate of gas production (k), but decreased the theoretical maximum of gas production ($V_F$) and the half-life ($t_{0.5}$) compared with the control. The $NH_3-N$ concentration reached the lowest concentration with 0.01 mg MCS/mL at 6 h. The MSC inclusion increased (p<0.001) the molar proportion of butyrate, isovalerate at 24 h and 48 h, and the molar proportion of acetate at 24 h, but then decreased (p<0.05) them at 48 h. The molar proportion of valerate was increased (p<0.05) at 24 h. The acetate to propionate ratio (A/P; linear, p<0.01) was increased at 24 h, but reached the least value at the level of 0.30 mg/mL MCS. The MCS inclusion decreased (p<0.05) the molar proportion of propionate at 24 h and then increased it at 48 h. The concentration of total volatile fatty acid was decreased (p<0.001) at 24 h, but reached the greatest concentration at the level of 0.01 mg/mL and the least concentration at the level of 0.60 mg/mL. The relative abundance of Ruminococcus albus was increased at 6 h and 24 h, and the relative abundance of Fibrobacter succinogenes was the lowest (p<0.05) at 0.60 mg/mL at 6 h and 24 h. The relative abundance of Butyrivibrio fibrisolvens and fungus reached the greatest value (p<0.05) at low doses of MCS inclusion and the least value (p<0.05) at 0.60 mg/mL at 24 h. The present results demonstrates that a high level of MCS quickly inhibits in vitro fermentation of maize stover, while MCS at low doses has the ability to modulate the ruminal fermentation pattern by regulating the number of functional rumen microbes including cellulolytic bacteria and fungi populations, and may have potential as a feed additive applied in the diets of ruminants.

• Journal of Animal Science and Technology
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• v.48 no.4
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• pp.521-532
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• 2006

An in vitro study was conducted to examine the effect of addition level of carbohydrates on fermentation characteristics, and bio-hydrogenation of unsaturated fatty acids by mixed rumen bacteria when incubated with soybean oil or flaxseed oil. Four levels(0%, 0.3%, 0.6% and 0.9%, w/v) of the mixed carbohydrates(glucose, cellobiose, soluble starch, 1:1:1, in weight basis) and oil sources(soybean or flaxseed oil, 60mg in 150ml culture solution) were added to the mixed solution of strained rumen fluid with artificial saliva(1:4, v/v), and incubated anaerobically for 12 hours at 39℃. pH and ammonia-N concentration were lower by increasing the substrate levels at all incubation periods(P<0.05～P<0.001). The propionate proportion increased(P<0.001), but acetic acid and butyric acid decreased(P<0.001) with the substrate level at 6 and 12 h incubations. Oil sources did not influence the proportions of individual VFA. At the end of incubation, the proportions of C18:0(P<0.01), C18:1(P<0.001) and trans-11C-18:1(P<0.01) were reduced but those of C18:2(P<0.001) and C18:3(P<0.01) were enhanced by the addition of flaxseed oil compared to addition of soybean oil. The proportions of C18:0 and total CLA were reduced(P<0.01) but those of trans-11-C18: (P<0.05) and C18:2(P<0.01) were increased with the substrate level when incubated with soybean oil or flaxseed oil. There were interactions(P<0.05) in the proportions of C18:1, C18:2 and C18:3(P<0.01) between oil source and substrate level. The proportions of cis-9, trans-11-CLA and trans-10, cis-12-CLA tended to reduce with substrate level, although there was no significant difference between treatments.

• Journal of Animal Science and Technology
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• v.48 no.5
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• pp.699-708
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• 2006

Ground rice, barley and corn were fed separately to the ruminally cannulated Hanwoo (Korean native cattle) for comparing their in situ and in vitro degradabilities, microbial growth, pH and gas production. It has been found that nearly all the dry matter (DM) and organic matter (OM) in barley and rice disappeared during 24 hr suspension in the rumen, but those in corn were only reduced by around 67%. Water soluble DM and OM fractions(‘a’), ranked from highest to lowest was corn, then rice and finally barley, but the order was reversed for content ‘b’, degradable fraction during time ‘t’. Judging by the degradation parameter of ‘b’ fraction, degradation rates per hour of DM and OM for barley were 38.3% and 37.2% respectively, significantly higher than those for rice (7.7% and 5.6%) and corn (4.1% and 1.3%). In general, results obtained from in vitro degradability of DM and OM were lower than those from in situ trials, but the ranking order of degradability was in agreement between both trials. In particular, ground rice has relatively lower in vitro microbial growth than corn or barley, but exhibited higher gas production. In addition, in vitro microbial growth of ground rice increased with up to 12 hr of incubation period, thereafter experienced a decrease with extended incubation time. pH of in vitro solution of rice decreased following 9 hr of incubation but gas production increased rapidly during the same period. From the results of DM and OM degradabilities and pH changes of in vitro solution with incubation time, it is concluded that rice represents a good source of energy for stability of rumen fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.10
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• pp.1449-1453
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• 2015

The objective of the present study was to determine the effects of feeding pelletized broccoli byproducts (PBB) on milk yield and milk composition in dairy cows. In Trial 1, an in vitro gas test determined the optimal replacement level of PBB in a concentrate mixture in a mixed substrate with Chinese wild ryegrass hay (50:50, w/w) at levels of 0, 10%, 20%, 30%, or 40% (dry matter basis). When the concentrate was replaced by PBB at a level of 20%, no adverse effects were found on the gas volume or its rate constant during ruminal fermentation. In trial 2, 24 lactating cows (days in milk = $170.4{\pm}35$; milk yield = $30{\pm}3kg/d$; body weight = $580{\pm}13kg$) were divided into 12 blocks based on day in milk and milk yield and randomly allocated to two dietary treatments: a basic diet with or without PBB replacing 20% of the concentrate mixture. The feeding trial lasted for 56 days; the first week allowed for adaptation to the diet. The milk composition was analyzed once a week. No significant difference in milk yield was observed between the two groups (23.5 vs 24.2 kg). A significant increase was found in milk fat content in the PBB group (p<0.05). Inclusion of PBB did not affect milk protein, lactose, total solids or solids-not-fat (p>0.05). These results indicated that PBB could be included in dairy cattle diets at a suitable level to replace concentrate mixture without any adverse effects on dairy performance.

• Korean Journal of Agricultural Science
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• v.29 no.2
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• pp.20-34
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• 2002

This study was conducted to examine the effects of an Aspergillus oryzae fermentation culture on the in vitro digestibilities of dry matter, crude fiber, acid detergent fiber (ADF), neutral detergent fiber (NDF), crude protein, and pH in in vitro experiment of diets for dairy cows. A fungal species, Aspergillus oryzae was supplied by Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea (KCTC 1229). The experimental diets were commercial compound feed (concentrate) and total mixed ration (TMR) for lactating cows, of which chemical analyses were determined at Research and Development Institute, Woosung Feed Co., Ltd., while the digestibilities were done at the laboratory of Chungnam National University. Aspergillus oryzae culture products were added to compound feed and TMR at the rate of 0, 1.0, 2.0, 3.0% respectively. The experimental diet with the rumen fluid sampled from Holstein fresian milking cows were used and digested for 24 hrs, 48hrs and 72hrs in the shaking incubator. The residues of the digesta were digested for 48hrs in the incubator in which put 30ml of 0.1N HCl with 0.2% pepsin at $39^{\circ}C$. The final precipitates were dried for 48hrs in the drier at $60^{\circ}C$. These experimental procedures were triplicated to determine the in vitro digestibility of dry matter, crude fiber, ADF, NDF, crude protein and pH. Compared to control diet, not added Aspergillus oryzae, the DM digestibility of fungal diets were improved 2.1%(63.1%), 9.7%(68.5%) and 9.0%(68.0%) for 24 hour fermentation in compound feed while 4.8%(60.0%), 6.4%(61.1%) and 2.9%(58.8%) in TMR. On the contrary, for 48 hour and 72 hour digestibilities, the effects of Aspergillus oryzae culture on the digestibility of dry matter were relatively lowered compared to 24 hour digestibility. Referring to the digestibility of dietary fiber, Aspergillus oryzae was believed to significantly improve digestibilities of crude fiber, ADF and NDF. Those were increased up to 13.3%(53.3%) for 24 hour fermentation, while 2.4%(54.6%) for 3.0% added for 72 hour fermentation in compound feed. However, there were no significant differences among the treatments for the inclusion rate of Aspergillus oryzae, even though the more inclusion rate, the better digestibility. The protein digestibilities were significantly improved from 0.4%(79.7%) to 9.4%(71.8%) by adding Aspergillus oryzae into compound feed. However, there were no significant differences between the two experimental diets, 2.0% and 3.0% Aspergillus oryzae included diets. In case of TMR, the protein digestibilities were significantly improved from 4.0%(70.4%) to 6.3%(65.1%) by adding Aspergillus oryzae. However, there were no significant differences between the two experimental diets, 2.0% and 3.0% Aspergillus oryzae included diets. In this study, there were no significant differences among the treatments in pH. On the contrary, there were slightly decrease in pH by adding Aspergillus oryzae into experimental diets but not significant. Summarizing the results of this examination, Aspergillus oryzae fermentation culture is believed to improve the digestibilities of dry matter, fiber and crude protein in cattle diets. However, more detailed research for the mechanism of the fungal culture is required to improve ruminal environment.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.9
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• pp.1296-1302
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• 2015

This study was conducted to examine the effects of supplementing brown seaweed by-products (BSB) in the diet of ruminants on ruminal fermentation characteristics, growth performance, endocrine response, and milk production in Holstein cows. In Experiment 1, the effects of different levels (0%, 2%, and 4% of basal diet as Control, 2% BSB, 4% BSB, respectively) of BSB were evaluated at 3, 6, 9, 12, and 24 h in vitro batch culture rumen fermentation. The pH tended to be higher for the higher level of BSB supplementation, with the pH at 12 h being significantly higher (p<0.05) than that of the control. The concentration of ammonia nitrogen was lower at 3, 9, 12, and 24 h incubation (p<0.05) compared with the control, and tended to be low at other incubation times. Volatile fatty acid concentration appeared to be minimally changed while lower values were observed with 4% BSB treatment at 24 h (p<0.05). In Experiment 2, effects of levels (0%, 2%, and 4%) of BSB on growth performance, endocrine responses and milk production were studied with Holstein dairy cows during transition. Dry matter intake, daily gain and feed efficiency were not affected by BSB supplementation. The concentration of plasma estrogen for the control, 2% BSB and 4% BSB after three months of pregnancy were 55.7, 94.1, and 72.3 pg/mL, respectively (p = 0.08). Although the differences of progesterone levels between BSB treatments and the control were minimal, the concentration in 4% BSB treatment increased to 157.7% compared with the initial level of the study. Triiodothyronine and thyroxine levels were also higher after both three months and eight months of pregnancy than the initial level at the beginning of the study. In addition, BSB treatments during one month after delivery did not affect daily milk yield and composition. In conclusion, the present results indicate that supplementation of BSB did not compromise ruminal fermentation, and animal performance at lower levels and hence may have potential to be used as a safe feed ingredient in dairy cows.