• Asian-Australasian Journal of Animal Sciences
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• v.32 no.12
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• pp.1864-1872
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• 2019

Objective: This study was conducted to evaluate the effects of Ecklonia stolonifera (E. stolonifera) extract addition on in vitro ruminal fermentation characteristics, methanogenesis and microbial populations. Methods: One cannulated Holstein cow ($450{\pm}30kg$) consuming timothy hay and a commercial concentrate (60:40, w/w) twice daily (09:00 and 17:00) at 2% of body weight with free access to water and mineral block were used as rumen fluid donors. In vitro fermentation experiment, with timothy hay as substrate, was conducted for up to 72 h, with E. stolonifera extract added to achieve final concentration 1%, 3%, and 5% on timothy hay basis. Results: Administration of E. stolonifera extract to a ruminant fluid-artificial saliva mixture in vitro increased the total gas production. Unexpectedly, E. stolonifera extracts appeared to increase both methane emissions and hydrogen production, which is contrasts with previous observations with brown algae extracts used under in vitro fermentation conditions. Interestingly, real-time polymerase chain reaction indicated that as compared with the untreated control the ciliate-associated methanogen and Fibrobacter succinogenes populations decreased, whereas the Ruminococcus flavefaciens population increased as a result of E. stolonifera extract supplementation. Conclusion: E. stolonifera showed no detrimental effect on rumen fermentation characteristics and microbial population. Through these results E. stolonifera has potential as a viable feed supplement to ruminants.

• Animal Bioscience
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• v.35 no.10
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• pp.1592-1605
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• 2022

Objective: The objective of this study was to select an effective in vitro digestion-fermentation model to estimate the effect of decreasing dietary crude protein (CP) on odor emission during pig production and to suggest potential prediction markers through in vitro and in vivo experiments. Methods: In the in vitro experiment, three diet formulations with different CP contents (170 g/kg, 150 g/kg, and 130 g/kg) but containing the same standardized ileal digestible essential amino acids (SID-EAA) were assessed. Each diet was evaluated by two different in vitro gastric-intestinal phase digestion methods (flask and dialysis), combined with fresh pig feces-ferment inoculation. Eighteen growing barrows (31.9±1.6 kg) were divided into three groups: control diet (180 g CP/kg, without SID-EAA adjustment), 170 g CP/kg diet, and 150 g CP/kg diet for 4 weeks. Results: The in vitro digestion results indicated that in vitro digestibility was affected by the gastric-intestinal phase digestion method and dietary CP level. According to the gas kinetic and digestibility results, the dialysis method showed greater distinguishability for dietary CP level adjustment. Nitrogen-related odor compounds (NH₃-N, indole, p-cresol, and skatole) were highly correlated with urease and protease activity. The feeding study indicated that both EAA-adjusted diets resulted in a lower odor emission especially in p-cresol and skatole. Both protease and urease activity in feces were also closely related to odor emissions from nitrogen metabolism compounds. Conclusion: Dialysis digestion in the gastric-intestinal phase followed by fresh fecal inoculation fermentation is suitable for in vitro diet evaluation. The enzyme activity in the fermentation and the fecal samples might provide a simple and effective estimation tool for nitrogen-related odor emission prediction in both in vitro and in vivo experiments.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.8
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• pp.1136-1144
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• 2016

Leucaena silage was supplemented with different levels of molasses and urea to study its nutritive value and in vitro rumen fermentation efficiency. The ensiling study was randomly assigned according to a $3{\times}3$ factorial arrangement in which the first factor was molasses (M) supplement at 0%, 1%, and 2% of crop dry matter (DM) and the second was urea (U) supplement as 0%, 0.5%, and 1% of the crop DM, respectively. After 28 days of ensiling, the silage samples were collected and analyzed for chemical composition. All the nine Leucaena silages were kept for study of rumen fermentation efficiency using in vitro gas production techniques. The present result shows that supplementation of U or M did not affect DM, organic matter, neutral detergent fiber, and acid detergent fiber content in the silage. However, increasing level of U supplementation increased crude protein content while M level did not show any effect. Moreover, the combination of U and M supplement decreased the content of mimosine concentration especially with M2U1 (molasses 2% and urea 1%) silage. The result of the in vitro study shows that gas production kinetics, cumulation gas at 96 h and in vitro true digestibility increased with the increasing level of U and M supplementation especially in the combination treatments. Supplementation of M and U resulted in increasing propionic acid and total volatile fatty acid whereas, acetic acid, butyric acid concentrations and methane production were not changed. In addition, increasing U level supplementation increased $NH_3$-N concentration. Result from real-time polymerase chain reaction revealed a significant effect on total bacteria, whereas F. succinogenes and R. flavefaciens population while R. albus was not affected by the M and U supplementation. Based on this study, it could be concluded that M and urea U supplementation could improve the nutritive value of Leucaena silage and enhance in vitro rumen fermentation efficiency. This study also suggested that the combination use of M and U supplementation level was at 2% and 1%, respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.4
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• pp.496-501
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• 2012

Thirty-five available feeds were fermented in vitro in order to investigate their soluble total organic carbon (TOCs) and methane ($CH_4$) production rate. A fermentation reactor was designed to capture the $CH_4$ gas emitted and to collect liquor from the reactor during in vitro fermentation. The results showed that $CH_4$ production rate greatly varied among feeds with different ingredients. The lowest $CH_4$-producing feeds were corn gluten feed, brewer's grain, and orchard grass among the energy, protein, and forage feed groups, respectively. Significant differences (p<0.05) were found in digestibility, soluble total organic carbon (TOCs), and $CH_4$ emissions among feeds, during 48 h of in vitro fermentation. Digestibility and TOCs was not found to be related due to different fermentation pattern of each but TOCs production was directly proportional to $CH_4$ production (y = 0.0076x, $r^2$ = 0.83). From this in vitro study, TOCs production could be used as an indirect index for estimation of $CH_4$ emission from feed ingredients.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.11
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• pp.1571-1576
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• 2014

Nutritive values of green and black tea by-products and anti-nutritive activity of their tannins were evaluated in an in vitro rumen fermentation using various molecular weights of polyethylene glycols (PEG), polyvinyl pyrrolidone (PVP) and polyvinyl polypyrrolidone as tannin-binding agents. Significant improvement in gas production by addition of PEG4000, 6000 and 20000 and PVP was observed only from black tea by-product, but not from green tea by-product. All tannin binding agents increased $NH_3$-N concentration from both green and black tea by-products in the fermentation medium, and the PEG6000 and 20000 showed relatively higher improvement in the $NH_3$-N concentration. The PEG6000 and 20000 also improved in vitro organic matter digestibility and metabolizable energy contents of both tea by-products. It was concluded that high molecular PEG would be suitable to assess the suppressive activity of tannins in tea by-products by in vitro fermentation. Higher responses to gas production and $NH_3$-N concentration from black tea by-product than green tea by-product due to PEG indicate that tannins in black tea by-product could suppress rumen fermentation more strongly than that in green tea by-product.

• Journal of the Korea Organic Resources Recycling Association
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• v.8 no.4
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• pp.100-110
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• 2000

Studies were conducted to know effects of the bulking agents (saw dusts, mushroom waste, wheat bran coconut meal, rice hulls) adding o moisture control, fermentation methods (aerobic and anaerobic) and periods (1 to 20 days) of food waste fermentation for animal feeds on chemical compositions and in vitro DDM (digestibility of dry matter). Experiment designs were focussed basically to obtain extension service data. The NDF (neutral detergent fiber) composition in the oak and pine saw dust were 93.5% and 95.4% (DM basis) in respectively. Thus, the fermented food waste feeds using saw dust (50%) increased NDF(12%), and decreased in vitro DDM(48%) compared to those of raw materials before aerobic fermentation. The oak saw dust showed higher DDM compared to pine. Mushroom wastes which is a residues of mushroom culture mixed originally willow saw dust (80%) and wheat bran (20%) showed quite higher feed value compared to both saw dusts. It was found that an in vitro DDM and NDF composition in fermented feeds appeared highly dependent or the NDF composition in bulking agents. With an increase wheat bran ratio substitute mushroom waste showed linearly decreased NDF, and increased in vitro DDM in the fermented food waste feeds. The fermented feeds added bottling agents composed higher NDF resulted in higher NDF and lower in vitro DDM with prolonged fermentation time. The feeds from anaerobic fermentation appeared lower NDF and higher in vitro DDM compared to those of aerobic fermentation.

• Korean Journal of Agricultural Science
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• v.45 no.3
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• pp.419-427
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• 2018

The present study was conducted to investigate the effect of different levels of Korean corn grain on in vitro ruminal fermentation with total mixed ration (TMR) as a basal feed. Three ruminal cannulated Holstein steers (Body Weight $479{\pm}33.0kg$) were used as rumen fluid donors. Treatments for in vitro fermentation were TMR only (control, 3.0 g), TMR substituted partially with high level (HC, TMR 1.5 and corn 1.5 g), and with low level of Korean corn grain (LC, TMR 2.25 and corn 0.75 g), respectively. To measure in vitro ruminal pH, gas production, ammonia N and volatile fatty acids (VFA), the in vitro fermentation incubation was triplicated at $39^{\circ}C$, 120 rpm for 0, 1, 3, 6, 12, 24 and 48 h, respectively. Mean ruminal pH was significantly lower (p < 0.05) for HC than control. Changes in rumen pH was rather similar between the groups till 6 h after incubation, but the lowest pH for HC (pH 5.10) appeared at 48 h compared with control and LC. Total gas production was tended (p < 0.09) to be higher and ammonia N was significantly lower (p < 0.05) for HC than control and LC. Total VFA was higher (p < 0.05) for HC and LC than control but no differences appeared between HC and LC. Overall, the present data indicate that feeding different levels of Korean domestic corn grain may lead to high and sustainable starch degradation in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.470-478
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• 2017

Objective: This study aims to evaluate the chemical composition, fermentation quality and in vitro ruminal fermentation of various ratios and storage periods of liquid brewer's yeast (LBY) mixed with cassava pulp (CVP). Methods: Four mixtures of fresh LBY and CVP were made (LBY0, LBY10, LBY20, and LBY30 for LBY:CVP at 0:100, 10:90, 20:80, and 30:70, respectively) on a fresh matter basis, in 500 g in plastic bags and stored at 30 to $32^{\circ}C$. After storage, the bags were opened weekly from weeks 0 to 4. Fermentation quality and in vitro gas production (IVGP) were determined, as well as the dry matter (DM), organic matter (OM), crude protein (CP), ether extract (EE), neutral detergent fiber, acid detergent fiber and acid detergent lignin contents. Results: The contents of CP and EE increased, whereas all other components decreased, in proportion to LBY inclusion (p<0.01). The DM and OM contents gradually decreased in weeks 3 and 4 (p<0.05), while EE contents were lowest in week 0. The pH, ammonia nitrogen per total nitrogen ($NH_3-N/TN$) and V-score in each mixture and storage period demonstrated superior fermentation quality ($pH{\leq}4.2$, $NH_3-N/TN{\leq}12.5%$, and V-score>90%). The pH increased and $NH_3-N/TN$ decreased, with proportionate increases of LBY, whereas the pH decreased and $NH_3-N/TN$ increased, as the storage periods were extended (p<0.01). Although IVGP decreased in proportion to the amount of LBY inclusion (p<0.01), in vitro organic matter digestibility (IVOMD) was unaffected by the mixture ratios. The highest IVGP and IVOMD were observed in week 0 (p<0.01). Conclusion: The inclusion of LBY (as high as 30%) into CVP improves the chemical composition of the mixture, thereby increasing the CP content, while decreasing IVGP, without decreasing fermentation quality and IVOMD. In addition, a preservation period of up to four weeks can guarantee superior fermentation quality in all types of mixtures. Therefore, we recommend limiting the use of CVP as a feed ingredient, given its low nutritional value and improving feed quality with the inclusion of LBY.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.6
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• pp.797-805
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• 2014

The effect of concentrate mixtures with crude protein (CP) levels 10%, 13%, 16%, and 19% and diets with roughage to concentrate ratios 80:20, 60:40, 40:60, and 20:80 (w/w) were determined on dry matter (DM) and organic matter (OM) digestibility, and fermentation metabolites using an in vitro fermentation technique. In vitro fermented attributes were measured after 4, 24, and 48 h of incubation respectively. The digestibility of DM and OM, and total volatile fatty acid (VFA) increased whereas pH decreased with the increased amount of concentrate in the diet (p<0.001), however CP levels of concentrate did not have any influence on these attributes. Gas production reduced with increased CP levels, while it increased with increasing concentrate levels. Ammonia nitrogen ($NH_3$-N) concentration and microbial CP production increased significantly (p<0.05) by increasing CP levels and with increasing concentrate levels in diet as well, however, no significant difference was found between 16% and 19% CP levels. Therefore, 16% CP in concentrate and increasing proportion of concentrate up to 80% in diet all had improved digestibility of DM and organic matter, and higher microbial protein production, with improved fermentation characteristics.

• Journal of Microbiology and Biotechnology
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• v.26 no.3
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• pp.558-566
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• 2016

Biotic agents such as fumarate-reducing bacteria can be used for controlling methane (CH₄) production in the rumen. Fumarate-reducing bacteria convert fumarate to succinate by fumarate reductase, ultimately leading to the production of propionate. Fumarate-reducing bacteria in the genus Enterococcus were isolated from rumen fluid samples from slaughtered Korean native goats. The enterococci were identified as Enterococcus faecalis SROD5 and E. faecium SROD by phylogenetic analyses of 16S rRNA gene sequences. The fumarate reductase activities of the SROD5 and SROD strains were 42.13 and 37.05 mM NADH oxidized/min/mg of cellular nitrogen (N), respectively. Supplementation of rumen fermentation in vitro with the SROD5 and SROD strains produced significantly higher propionate, butyrate, and total volatile fatty acid (VFA) concentrations than controls at 12 h; VFA concentrations tended to increase after 24 h of incubation. The generated CH₄ concentration was significantly lower in the SROD5 and SROD treatment groups after 24 h of incubation. These findings indicate that E. faecium SROD has potential as a direct-fed microbial additive for increasing total VFAs while decreasing CH₄ production in rumen fermentation in vitro.