• Microbiology and Biotechnology Letters
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• v.42 no.1
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• pp.32-40
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• 2014

Methane oxidation and the production potentials of ground soil (soil A) and garden soil (soil B, C, & D) in an urban school were evaluated, and the methanotrophic and methanogen communities in the soil samples were quantified using quantitative realtime PCR. The methanotrophic community in the raw soil A sample possessed a $6.1{\times}10^3$ gene copy number/g dry weight soil, whereas those in the raw soils B~D samples were $1.6-1.9{\times}10^5$ gene copy numbers/g dry weight soil. Serum bottles added with the soil samples were enriched with methane gas, and then evaluated for their methane oxidation potential. The soil A sample had a longer induction phase for methane oxidation than the other soils. However, soil A showed a similar methane oxidation potential with soils B~D after the induction phase. The methanotrophic community in the enriched soil A sample was increased by up to $2.3{\times}10^7$ gene copy numbers/g dry weight soil, which had no significantly difference compared with those in soils B~D ($1.2-2.8{\times}10^8$ gene copy numbers/g dry weight soil). Methane production showed a similar tendency to methane oxidation. The methanogens community in raw soil A ($1.7{\times}10^5$ gene copy number/g dry weight soil) was much less than those in raw soils B~D ($1.3-3.4{\times}10^7$ gene copy numbers/g dry weight soil). However, after methane gas was produced by adding starch to the soils, soil samples A~D showed $10^7$ gene copy numbers/g dry weight soil in methanogens communities. The results indicate that methanotrophic and methanogenic bacteria have coexisted in this urban school's soils. Moreover, under appropriate conditions for methane oxidation and production, methanotrophic bacteria and methanogens are increased and they have the potential for methane oxidation and production.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.1
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• pp.36-45
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• 2014

The objective of this study was to determine the effects of protein sources and roughage (R) to concentrate (C) ratio on in vitro fermentation parameters using a gas production technique. The experimental design was a $2{\times}5$ factorial arrangement in a completely randomized design (CRD). Factor A was 2 levels of protein sources yeast fermented cassava chip protein (YEFECAP) and soybean meal (SBM) and factor B was 5 levels of roughage to concentrate (R:C) ratio at 80:20, 60:40, 40:60, 20:80, and 0:100, respectively. Rice straw was used as a roughage source. It was found that gas production from the insoluble fraction (b) of YEFECAP supplemented group was significantly higher (p<0.05) than those in SBM supplemented group. Moreover, the intercept value (a), gas production from the insoluble fraction (b), gas production rate constants for the insoluble fraction (c), potential extent of gas production (a+b) and cumulative gas production at 96 h were influenced (p<0.01) by R:C ratio. In addition, protein source had no effect (p>0.05) on ether in vitro digestibility of dry matter (IVDMD) and organic (IVOMD) while R:C ratio affected the IVDMD and IVOMD (p<0.01). Moreover, YEFECAP supplanted group showed a significantly increased (p<0.05) total VFA and $C_3$ while $C_2$, $C_2:C_3$ and $CH_4$ production were decreased when compared with SBM supplemented group. In addition, a decreasing R:C ratio had a significant effect (p<0.05) on increasing total VFA, $C_3$ and $NH_3$-N, but decreasing the $C_2$, $C_2:C_3$ and CH4 production (p<0.01). Furthermore, total bacteria, Fibrobacter succinogenes, Ruminococcus flavefaciens and Ruminococcus albus populations in YEFECAP supplemented group were significantly higher (p<0.05) than those in the SBM supplemented group while fungal zoospores, methanogens and protozoal population remained unchanged (p>0.05) as compared between the two sources of protein. Moreover, fungal zoospores and total bacteria population were significantly increased (p<0.01) while, F. succinogenes, R. flavefaciens, R. albus, methanogens and protozoal population were decreased (p<0.01) with decreasing R:C ratio. In conclusion, YEFECAP has a potential for use as a protein source for improving rumen fermentation efficiency in ruminants.

• Korean Journal of Environmental Agriculture
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• v.33 no.4
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• pp.282-289
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• 2014

BACKGROUND: Methane($CH_4$) is considered as the secondmost potent greenhouse gas after carbon dioxide ($CO_2$). Methanogenesis is an enzyme-mediated multi-step process by methanogens. In the penultimate step, methylated Co-M is reduced by methyl Co-M reductase (MCR) to $CH_4$ involving a nickel-containing cofactor F430. The activity of MCR enzyme is dependent on the F430 and therefore, the bioavailability of Ni to methanogens is expected to influence MCR activity and $CH_4$ production in soil. In this study, different doses of EDTA(Ethylene Diamine Tetraacetic Acid) were applied in flooded soils to evaluate their suppression effect on methane production by chelating Ni of methanogenesis cofactor. METHODS AND RESULTS: EDTA was selected as chelating agents and added into wetland and rice paddy soil at the rates of 0, 25, 50, 75, and $100mmol\;kg^{-1}$ before 4-weeks incubation test. During the incubation, cumulative $CH_4$ production patterns were characterized. At the end of the experiment, soil samples were removed from their jars to analyze total soil Ni and water-soluble Ni content and methanogen abundance. Methane production from 100 mmol application decreased by 55 and 78% in both soils compared to that from 0 mmol. With increasing application rate of EDTA in both soils, water-soluble Ni concentration significantly increased, but total soil Ni and methanogen activities showed negative relationship during incubation test. CONCLUSION: The decrease in methane production with EDTA application was caused by chelating Ni of coenzyme F430 and inhibiting methanogenesis by methyl coenzyme M reductase. Consequently, EDTA application decreased uptake of Ni into methanogen, subsequently inhibited methanogen activities and reduced methane production in flooded soils.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.7
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• pp.312-320
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• 2020

The objective of the study was to undertake a phylogenetic diversity census of ruminal archaea based on a meta-analysis of 16S rRNA gene sequences that were publicly available in the Ribosomal Database Project. A total of 8,416 sequences were retrieved from the Ribosomal Database Project (release 11, update 5) and included in the construction of a taxonomy tree. Species-level operational taxonomic units (OTUs) were analyzed at a 97% sequence similarity by using the QIIME program. Of the 8,416 sequences, 8,412 were classified into one of three phyla; however, the remaining four sequences could not be classified into a known phylum. The Euryarchaeota phylum was predominant and accounted for 99.8% of the archaeal sequences examined. Among the Euryarchaeota, 65.4% were assigned to Methanobrevibacter, followed by Methanosphaera (10.4%), Methanomassillicoccus (10.4%), Methanomicrobium (7.9%), Methanobacterium (1.9%), Methanimicrococcus (0.5%), Methanosarcina (0.1%), and Methanoculleus (0.1%). The 7,544 sequences that had been trimmed to the V2 and V3 regions clustered into 493 OTUs. Only 17 of those 493 OTUs were dominant groups and accounted for more than 1% of the 7,544 sequences. These results can help guide future research into the dominant ruminal methanogens that significantly contribute to methane emissions from ruminants, research that may lead to the development of anti-methanogenic compounds that inhibit these methanogens regardless of diet or animal species.

• Journal of the Korea Organic Resources Recycling Association
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• v.14 no.3
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• pp.126-137
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• 2006

Behaviors of simple organic compound and granular sludge in an upflow anaerobic sludge blanket (UASB) reactor treating propionate at high ammonia nitrogen levels were investigated for 12 months. The UASB reactor achieved about 80% removal of chemical oxygen demand (COD) at ammonia nitrogen concentration up to 6000 mg-N/L. At higher concentration of ammonia nitrogen, the propionate in the effluent increased whereas the acetate was very low. At ammonia nitrogen concentration of 8000 mg-N/L, the volatile suspended solids (VSS) increased sharply due probably to the decrease of the content of extracellular polymer (ECP) although methane production was very low. The specific methanogenic activity (SMA) using formate, acetate, and propionate as substrate to granules decreased as ammonia nitrogen concentration increased. The ammonia nitrogen concentration $I^{50}$, causing 50% inhibition of SMA were 2666, 4778 and 5572 mg-N/L, respectively. The kinetic coefficients of ammonia inhibition using formate, acetate, and propionate as substrate were 3.279, 0.999 and 0.609, respectively. The SMA using formate was severely affected by ammonia nitrogen than those using acetate and propionate. This result indicated that the hydrogenotrophic methanogens was most affected by ammonia nitrogen. Granules were mainly composed of microcolonies of methanothrix-like bacteria resembling bamboo-shape, and several other microcolonies including propionate degrader with juxtapositioned syntrophic associations between the hydrogen-producing acetogens and hydrogen-consuming methanogens.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.2
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• pp.285-294
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• 2011

The abatement of methane emission from ruminants is an important global issue due to its contribution to greenhouse gas with carbon dioxide. Methane is generated in the rumen by methanogens (archaea) that utilize metabolic hydrogen ($H_2$) to reduce carbon dioxide, and is a significant electron sink in the rumen ecosystem. Therefore, the competition for hydrogen used for methanogenesis with alternative reductions of rumen microbes should be an effective option to reduce rumen methanogenesis. Some methanogens parasitically survive on the surface of ciliate protozoa, so that defaunation or decrease in protozoa number might contribute to abate methanogenesis. The most important issue for mitigation of rumen methanogenesis with manipulators is to secure safety for animals and their products and the environment. In this respect, prophylactic effects of probiotics, prebiotics and miscellaneous compounds to mitigate rumen methanogenesis have been developed instead of antibiotics, ionophores such as monensin, and lasalocid in Japan. Nitrate suppresses rumen methanogenesis by its reducing reaction in the rumen. However, excess intake of nitrate causes intoxication due to nitrite accumulation, which induces methemoglobinemia. The nitrite accumulation is attributed to a relatively higher rate of nitrate reduction to nitrite than nitrite to ammonia via nitroxyl and hydroxylamine. The in vitro and in vivo trials have been conducted to clarify the prophylactic effects of L-cysteine, some strains of lactic acid bacteria and yeast and/or ${\beta}$1-4 galactooligosaccharide on nitrate-nitrite intoxication and methanogenesis. The administration of nitrate with ${\beta}$1-4 galacto-oligosaccharide, Candida kefyr, and Lactococcus lactis subsp. lactis were suggested to possibly control rumen methanogenesis and prevent nitrite formation in the rumen. For prebiotics, nisin which is a bacteriocin produced by Lactococcus lactis subsp. lactis has been demonstrated to abate rumen methanogenesis in the same manner as monensin. A protein resistant anti-microbe (PRA) has been isolated from Lactobacillus plantarum as a manipulator to mitigate rumen methanogenesis. Recently, hydrogen peroxide was identified as a part of the manipulating effect of PRA on rumen methanogenesis. The suppressing effects of secondary metabolites from plants such as saponin and tannin on rumen methanogenesis have been examined. Especially, yucca schidigera extract, sarsaponin (steroidal glycosides), can suppress rumen methanogenesis thereby improving protein utilization efficiency. The cashew nutshell liquid (CNSL), or cashew shell oil, which is a natural resin found in the honeycomb structure of the cashew nutshell has been found to mitigate rumen methanogenesis. In an attempt to seek manipulators in the section on methane belching from ruminants, the arrangement of an inventory of mitigation technologies available for the Clean Development Mechanism (CDM) and Joint Implementation (JI) in the Kyoto mechanism has been advancing to target ruminant livestock in Asian and Pacific regions.

• Journal of the Korea Organic Resources Recycling Association
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• v.7 no.2
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• pp.25-34
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• 1999

The effects of copper on the anaerobic degradation of propionate were studied using anaerobic batch reactors. The apparent inhibitory effects of copper on the anaerobic degradation of propionate could be observed from behaviors of intermediates, ultimate methane yield(UMY) and specific methanogenic activity(SMA) There was little inhibition at the concentration of $2.5mg\;Cu^{2+}/L$. Beyond this concentration, the inhibitory effects increased with increasing dose of coppers. The 50% inhibition of UMY and SMA occurred at copper dosage of 33.8 and $24.1mg\;Cu^{2+}/gVSS$, respectively. The inhibitory effect based on the UMY was gradually reduced with the operation time dueprobably to the acclimation of microorganisms and/or binding of the added copper by ligands(and possibly ion exchange sites)contained on the cell membrane and extracellular polymer matrix whereas it based on the SMA might exclude the this phenomena. Therefore, the methodology for interpretation of inhibition data based on the SMA was more accurated than the UMY. There was no inhibitory effect in batch reactors supplemented with sulfate due to an antagonistic action of the sulfate reducing bacteria. Propionate degradation was initially retarded for copper inhibited samples but it gradually degraded afterward. Based on the mass removal considering take into account the propionate to acetate conversion, propionate degradation may appeal more affected than acetate. This result revealed that the hydrogenotrophic methanogens were the most affected by copper.

• Journal of Animal Science and Technology
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• v.65 no.1
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• pp.132-148
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• 2023

Ruminants are the main contributors to methane (CH₄), a greenhouse gas emitted by livestock, which leads to global warming. In addition, animals experience heat stress (HS) when exposed to high ambient temperatures. Organic trace minerals are commonly used to prevent the adverse effects of HS in ruminants; however, little is known about the role of these minerals in reducing enteric methane emissions. Hence, this study aimed to investigate the influence of dietary organic trace minerals on rumen fermentation characteristics, enteric methane emissions, and the composition of rumen bacteria and methanogens in heat-stressed dairy steers. Holstein (n=3) and Jersey (n=3) steers were kept separately within a 3×3 Latin square design, and the animals were exposed to HS conditions (Temperature-Humidity Index [THI], 82.79 ± 1.10). For each experiment, the treatments included a Control (Con) consisting of only basal total mixed rations (TMR), National Research Council (NRC) recommended mineral supplementation group (NM; TMR + [Se 0.1 ppm + Zn 30 ppm + Cu 10 ppm]/kg dry matter), and higher concentration of mineral supplementation group (HM; basal TMR + [Se 3.5 ppm + Zn 350 ppm + Cu 28 ppm]/kg dry matter). Higher concentrations of trace mineral supplementation had no influence on methane emissions and rumen bacterial and methanogen communities regardless of breed (p > 0.05). Holstein steers had higher ruminal pH and lower total volatile fatty acid (VFA) concentrations than Jersey steers (p < 0.05). Methane production (g/d) and yield (g/kg dry matter intake) were higher in Jersey steers than in Holstein steers (p < 0.05). The relative abundances of Methanosarcina and Methanobrevibacter olleyae were significantly higher in Holstein steers than in Jersey steers (p < 0.05). Overall, dietary organic trace minerals have no influence on enteric methane emissions in heat-stressed dairy steers; however, breed can influence it through selective alteration of the rumen methanogen community.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.521-524
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• 2001

The anaerobic continuous reactor, which was filled with a sludge of anaerobic digestion from Sooyoung sewage treatment plant, was supplied with synthetic wastewater of various concentration. After changing to substrate concentration, 디 1is research indicated that attached biomass was kept constant after attachment 23 days. In SEM photographs. shape and structure of biofilm could be observed, but bacteria species and methanogens were not identified. A large number of methanogenic bacteria were showed on the surface of PE substratum by fluorescence under 480nm of radiation.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.6
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• pp.700-707
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• 2010

Effects of protease-resistant antimicrobial substances (PRA) produced by Lactobacillus plantarum and Leuconostoc citreum on rumen methanogenesis were examined using the in vitro continuous methane quantification system. Four different strains of lactic acid bacteria, i) Lactococcus lactis ATCC19435 (Control, non-antibacterial substances), ii) Lactococcus lactis NCIMB702054 (Nisin-Z), iii) Lactobacillus plantarum TUA1490L (PRA-1), and iv) Leuconostoc citreum JCM9698 (PRA-2) were individually cultured in GYEKP medium. An 80 ml aliquot of each supernatant was inoculated into phosphate-buffered rumen fluid. PRA-1 remarkably decreased cumulative methane production, though propionate, butyrate and ammonia N decreased. For PRA-2, there were no effects on $CH_4$ and $CO_2$ production and fermentation characteristics in mixed rumen cultures. The results suggested that PRA-1 reduced the number of methanogens or inhibited utilization of hydrogen in rumen fermentation.