• Animal Bioscience
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• v.37 no.2_spc
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• pp.360-369
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• 2024

Ruminal methane production functions as the main sink for metabolic hydrogen generated through rumen fermentation and is recognized as a considerable source of greenhouse gas emissions. Methane production is a complex trait affected by dry matter intake, feed composition, rumen microbiota and their fermentation, lactation stage, host genetics, and environmental factors. Various mitigation approaches have been proposed. Because individual ruminants exhibit different methane conversion efficiencies, the microbial characteristics of low-methane-emitting animals can be essential for successful rumen manipulation and environment-friendly methane mitigation. Several bacterial species, including Sharpea, uncharacterized Succinivibrionaceae, and certain Prevotella phylotypes have been listed as key players in low-methane-emitting sheep and cows. The functional characteristics of the unclassified bacteria remain unclear, as they are yet to be cultured. Here, we review ruminal methane production and mitigation strategies, focusing on rumen fermentation and the functional role of rumen microbiota, and describe the phylogenetic and physiological characteristics of a novel Prevotella species recently isolated from low methane-emitting and high propionate-producing cows. This review may help to provide a better understanding of the ruminal digestion process and rumen function to identify holistic and environmentally friendly methane mitigation approaches for sustainable ruminant production.

• Journal of Animal Science and Technology
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• v.59 no.6
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• pp.14.1-14.9
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• 2017

Background: Garlic and its constituents are reported to have been effective in reducing methane emission and also influence glucose metabolism in body; however, studies in ruminants using garlic leaves are scarce. Garlic leaves contain similar compounds as garlic bulbs, but are discarded in field after garlic bulb harvest. We speculate that feeding garlic leaves might show similar effect as garlic constituents in sheep and could be potential animal feed supplement. Thus, we examined the effect of freeze dried garlic leaves (FDGL) on rumen fermentation, methane emission, plasma glucose kinetics and nitrogen utilization in sheep. Methods: Six sheep were fed Control diet (mixed hay and concentrate (60:40)) or FDGL diet (Control diet supplemented with FDGL at 2.5 g/kg $BW^{0.75}$ of sheep) using a crossover design. Methane gas emission was measured using open-circuit respiratory chamber. Plasma glucose turnover rate was measured using isotope dilution technique of [$U-^{13}C$]glucose. Rumen fluid, feces and urine were collected to measure rumen fermentation characteristics and nitrogen utilization. Result: No significant difference in rumen fermentation parameters was noticed except for rumen ammonia tended to be higher (0.05 < P < 0.1) in FDGL diet. Methane emission per kg dry matter ingested and methane emission per kg dry matter digested were lower (P < 0.05) in FDGL diet. Plasma glucose concentration was similar between diets and plasma glucose turnover rate tended to be higher in FDGL diet (0.05 < P < 0.1). Nitrogen retention was higher (P < 0.05) and microbial nitrogen supply tended to be higher (0.05 < P < 0.1) in FDGL diet. Conclusion: FDGL diet did not impair rumen fermentation, improved nitrogen retention; while absence of significant results in reduction of methane emission, glucose turnover rate and microbial nitrogen supply, further studies at higher dose would be necessary to conclude the merit of FDGL as supplement in ruminant feedstuff.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.7
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• pp.951-957
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• 2015

Four rumen fistulated swamp buffaloes were randomly assigned according to a $4{\times}4$ Latin square design to investigate the effects of Eucalyptus (E. Camaldulensis) leaf meal (ELM) supplementation as a rumen enhancer on feed intake and rumen fermentation characteristics. The dietary treatments were as follows: T1 = 0 g ELM/hd/d; T2 = 40 g ELM/hd/d; T3 = 80 g ELM/hd/d; T4 = 120 g ELM/hd/d, respectively. Experimental animals were kept in individual pens and concentrate was offered at 0.3% BW while rice straw was fed ad libitum. The results revealed that voluntary feed intake and digestion coefficients of nutrients were similar among treatments. Ruminal pH, temperature and blood urea nitrogen concentrations were not affected by ELM supplementation; however, ELM supplementation resulted in lower concentration of ruminal ammonia nitrogen. Total volatile fatty acids, propionate concentration increased with the increasing level of EML (p<0.05) while the proportion of acetate was decreased (p<0.05). Methane production was linearly decreased (p<0.05) with the increasing level of ELM supplementation. Protozoa count and proteolytic bacteria population were reduced (p<0.05) while fungal zoospores and total viable bacteria, amylolytic, cellulolytic bacteria were unchanged. In addition, nitrogen utilization and microbial protein synthesis tended to increase by the dietary treatments. Based on the present findings, it is suggested that ELM could modify the rumen fermentation and is potentially used as a rumen enhancer in methane mitigation and rumen fermentation efficiency.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.8
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• pp.1086-1091
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• 2011

This experiment was designed to investigate the effect of lerak extract on the dynamic of rumen microbes in the in vitro fermentation of diet with different ratios of forage and concentrate. In vitro fermentation was conducted according to the method of Tilley and Terry (1963). The design of experiment was a factorial block design with 2 factors. The first factor was the ratio of forage and concentrate (90:10, 80:20, and 70:30 w/w) and the second factor was the level of lerak extract (0, 0.6, and 0.8 mg/ml). Total volatile fatty acid (VFA) concentration, proportional VFA and NH3 concentration were measured at 4 h incubation. Protozoal numbers in the buffered rumen fluid after 4 and 24 h of incubation were counted under a microscope. Bacterial DNAs of buffered rumen fluid were isolated from incubated samples after 24 h of incubation using a QiaAmp kit. Total bacteria, Fibrobacter succinogenes, Ruminococcus albus, and Prevotella ruminicola were quantified using real time polymerase chain reaction (PCR). Lerak extract markedly reduced protozoal numbers in buffered rumen fluid of all diets after 24 h of incubation. Total bacteria did not change with lerak extract addition. While no difference in F. succinogenes was found, there was a slight increase in R. albus number and a significant enhancement in P. ruminicola number by increasing the level of lerak extract in all diets. Propionate concentration significantly increased in the presence of lerak extract at level 0.8 mg/ml. It was concluded that the addition of lerak extract could modify rumen fermentation and had positive effects on rumen microbes.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.3
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• pp.370-378
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• 2017

Objective: An experiment was conducted to investigate the effects of a specific mixture of essential oils (MEO), containing thyme, clove and cinnamon EO, on rumen microbial fermentation, nutrient apparent digestibility and blood metabolites in fistulated sheep. Methods: Six sheep fitted with ruminal fistulas were used in a repeated measurement design with two 24-d periods to investigate the effect of adding MEO at 0 (control), 0.8, and 1.6 mL/d on apparent nutrient digestibility, rumen fermentation characteristics, rumen microbial population and blood chemical metabolites. Animals were fed with a 50:50 alfalfa hay:concentrate diet. Results: Ruminal pH, total volatile fatty acids (VFA) concentration, molar proportion of individual VFA, acetate: propionate ratio and methane production were not affected with MEO. Relative to the control, Small peptides plus amino acid nitrogen and large peptides nitrogen concentration in rumen fluid were not affected with MEO supplementation; while, rumen fluid ammonia nitrogen concentration at 0 and 6 h after morning feeding in sheep fed with 1.6 mL/d of MEO was lower (p<0.05) compared to the control and 0.8 mL/d of MEO. At 0 h after morning feeding, ammonia nitrogen concentration was higher (p<0.05) in sheep fed 0.8 mL/d of MEO relative to 1.6 mL/d and control diet. Ruminal protozoa and hyper ammonia producing (HAP) bacteria counts were not affected by addition of MEO in the diet. Relative to the control, no changes were observed in the red and white blood cells, hemoglobin, hematocrit, glucose, beta-hydroxybutyric acid, cholesterol, total protein, albumin, blood urea nitrogen and aspartate aminotransferase and alanine aminotransferase concentration. Apparent total tract digestibility of dry matter, crude proten, organic matter, and neutral detergent fiber were not influenced by MEO supplementation. Conclusion:The results of the present study suggested that supplementation of MEO may have limited effects on apparent nutrient digestibility, ruminal fermentation and protozoa and HAP bacteria count, blood cells and metabolites.

• Animal Bioscience
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• v.37 no.4
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• pp.655-667
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• 2024

Objective: This study aimed to assess the impact of a hydroethanolic extract of walnut green husks (WGH) on rumen fermentation and the diversity of bacteria, methanogenic archaea, and fungi in sheep fed a high-concentrate diet. Methods: Five healthy small-tailed Han ewes with permanent rumen fistula were selected and housed in individual pens. This study adopted a self-controlled and crossover design with a control period and an experimental period. During the control period, the animals were fed a basal diet (with a ratio of concentrate to roughage of 65:35), while during the treatment period, the animals were fed the basal diet supplemented with 0.5% hydroethanolic extract of WGH. Fermentation parameters, digestive enzyme activities, and microbial diversity in rumen fluid were analyzed. Results: Supplementation of hydroethanolic extract of WGH had no significant effect on feed intake, concentrations of total volatile fatty acids, isovalerate, ammonia nitrogen, and microbial protein (p>0.05). However, the ruminal pH, concentrations of acetate, butyrate and isobutyrate, the ratio of acetate to propionate, protozoa count, and the activities of filter paper cellulase and cellobiase were significantly increased (p<0.05), while concentrations of propionate and valerate were significantly decreased (p<0.05). Moreover, 16S rRNA gene sequencing revealed that the relative abundance of rumen bacteria Christensenellaceae R7 group, Saccharofermentans, and Ruminococcaceae NK4A214 group were significantly increased, while Ruminococcus gauvreauii group, Prevotella 7 were significantly decreased (p<0.05). The relative abundance of the fungus Pseudomonas significantly increased, while Basidiomycota, Fusarium, and Alternaria significantly decreased (p<0.05). However, there was no significant change in the community structure of methanogenic archaea. Conclusion: Supplementation of hydroethanolic extract of WGH to a high-concentrate diet improved the ruminal fermentation, altered the structure of ruminal bacterial and fungal communities, and exhibited beneficial effects in alleviating subacute rumen acidosis of sheep.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.7
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• pp.1087-1095
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• 2020

Objective: The present study aimed to evaluate the effects of γ-aminobutyric acid (GABA)-producing bacteria (GPB) on in vitro rumen fermentation and on the growth performance and meat quality of Hanwoo steers. Methods: The effects of GPB (Lactobacillus brevis YM 3-30)-produced and commercially available GABA were investigated using in vitro rumen fermentation. Using soybean meal as a substrate, either GPB-produced or commercially available GABA were added to the in vitro rumen fermentation bottles, as follows: control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB; T3, 2 g/L autoclaved GPB; T4, 5 g/L autoclaved GPB; T5, 2 g/L GABA; and T6, 5 g/L GABA. In addition, 27 Hanwoo steers (602.06±10.13 kg) were subjected to a 129-day feeding trial, during which they were fed daily with a commercially available total mixed ration that was supplemented with different amounts of GPB-produced GABA (control, no additive; T1, 2 g/L GPB; T2, 5 g/L GPB). The degree of marbling was assessed using the nine-point beef marbling standard while endotoxin was analyzed using a Chromo-Limulus amebocyte lysate test. Results: In regard to in vitro rumen fermentation, the addition of GPB-produced GABA failed to significantly affect pH or total gas production but did increase the ammonia nitrogen (NH₃-N) concentration (p<0.05) and reduce total biogenic amines (p<0.05). Animals fed the GPB-produced GABA diet exhibited significantly lower levels of blood endotoxins than control animals and yielded comparable average daily gain, feed conversion ratio, and beef marbling scores. Conclusion: The addition of GPB improved in vitro fermentation by reducing biogenic amine production and by increasing both antioxidant activity and NH₃-N production. Moreover, it also reduced the blood endotoxin levels of Hanwoo steers.

• Korean Journal of Agricultural Science
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• v.48 no.4
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• pp.957-963
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• 2021

The study aimed to investigate the effects of supplemented sodium butyrate on the in vitro rumen fermentation and growth performance of Hanwoo calves. In total, four treatments were employed according to the sodium butyrate levels: no addition (control), an addition of 0.1% (treatment 1), an addition of 0.3% (treatment 2), and an addition of 0.5% (treatment 3). After 48 hours of fermentation, the ruminal pH was found to be higher in T1 than in C. Total volatile fatty acids were significantly higher in T2 and T3 than in C. The ratio of acetate and propionate was significantly lower in T1 and T3 than in C. In this study, the optimal concentration to promote rumen fermentation was found to be 0.3%, i.e., T2, and an experiment on Hanwoo calves at a farm was conducted. However, there were no significant differences between the treatment groups in terms of the daily weight gain, feed conversion ratio, and final body weight in the feeding experiment. Also, there were no significant differences in the body length, withers height, and height at hip cross between the control and the treatment groups. The addition of 0.3% sodium butyrate was most effective at promoting in vitro rumen fermentation, but it did not significantly affect the growth performance when fed to Hanwoo calves. This indicates that the addition of sodium butyrate improved rumen fermentation but did not have a growth-promoting effect. Future studies need to compare growth and carcass performance outcomes to confirm long-term effects.

• Journal of Animal Science and Technology
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• v.60 no.6
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• pp.15.1-15.10
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• 2018

Methane emission from the enteric fermentation of ruminant livestock is a main source of greenhouse gas (GHG) emission and a major concern for global warming. Methane emission is also associated with dietary energy lose; hence, reduce feed efficiency. Due to the negative environmental impacts, methane mitigation has come forward in last few decades. To date numerous efforts were made in order to reduce methane emission from ruminants. No table mitigation approaches are rumen manipulation, alteration of rumen fermentation, modification of rumen microbial biodiversity by different means and rarely by animal manipulations. However, a comprehensive exploration for a sustainable methane mitigation approach is still lacking. Dietary modification is directly linked to changes in the rumen fermentation pattern and types of end products. Studies showed that changing fermentation pattern is one of the most effective ways of methane abatement. Desirable dietary changes provide two fold benefits i.e. improve production and reduce GHG emissions. Therefore, the aim of this review is to discuss biology of methane emission from ruminants and its mitigation through dietary manipulation.