• Title/Summary/Keyword: In vitro gas production

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Effects of Addition Levels of Coffee and Green Tea By-products Extract including Polyphenols on in vitro Rumen Fermentation and Methane Emission (폴리페놀을 다량 함유한 커피박 및 녹차박 추출물의 수준별 첨가가 반추위 발효 및 메탄 발생량에 미치는 영향)

  • Won, Miyoung;Ryu, Chae-Hwa;Bak, Hyeryeon;Chae, Byungho;Jang, Seung-Ho;Choi, Seung-Shin;Choi, Bong-Hwan;Lee, Sung-Soo;Lee, Jinwook;Choi, Nag-Jin
    • Korean Journal of Organic Agriculture
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    • v.29 no.4
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    • pp.613-623
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    • 2021
  • This study was conducted to investigate the effect of addition levels of coffee and green tea by products extract including polyphenols through hot water extraction on rumen fermentation. The treatment groups consisted of coffee extract (CO), green tea extract (GR) and mixed extract (MIX), and the addition level was 10 µL, 20 µL and 30 µL of three levels. The experiment consisted of a total of 10 experimental groups including the control group, and a full factorial design was used. The effect of polyphenol addition in coffee and green tea by-products was analyzed through main and interaction effect of statistical analysis. The total polyphenol content of the extracts was 106.15, 79.10 and 185.25 ㎍ GAE/g DM for coffee by-product, green tea by-product and mixture, respectively. Total gas production was significantly lower in the treatment groups than in the control (114.00 mL/gDM) (p<0.05). Methane emission tended to decrease as the polyphenol addition level increased. Moreover, the MIX showed the lowest methane emission when 30 µL was added (p<0.05). Volatile fatty acids showed a significant difference compared to the treatment group as a control (98.06 mM) (p<0.05), but there was no change according to the level of polyphenols. As a result of the main effect and interaction, it is thought that the effect on methane reduction and improvement of rumen fermentation in MIX20 can be expected. In a series of studies, the addition of 20 µL of a blended extract of coffee and green tea by-products is thought to reduce methane to levels that do not inhibit rumen fermentation.

In vitro Evaluation of the Mechanism of Antagonism and Phosphate Solubilization by the Insect Gut Bacteria Pseudomonas sp. PRGB06 that Exhibits Plant Growth Promotion and Bio-Fertilizing Traits (배추좀나방 내장에서 분리한 식물생장촉진미생물 Pseudomonas sp. PRGB06의 길항기작과 인산가용화의 기내 평가)

  • Kim, Kyoung-A;Indiragandhi, P.;Anandham, R.;Palaniappan, P.;Trivedi, P.;Madhaiyan, M.;Han, Gwang-Hyun;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.41 no.1
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    • pp.18-25
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    • 2008
  • Pseudomonas sp. PRGB06, a bacterial strain isolated from diamondback moth (Plutella xylostella) gut, was examined for its plant growth promotion and biofertilizing traits. The bacteria growth was observed under various conditions of carbon sources, temperature, pH and salt concentrations. In addition, the mechanisms of antagonism and phosphate solubilization were investigated. The bacterial strain PRGB06, grew well using most of the tested carbon sources. The best growth was observed at $30^{\circ}C$ and pH 7. The inhibition of the pathogenic fungi was likely due to the volatile antifungal metabolite and ammonia gas produced by the bacteria. A significant positive relationship was found between the phosphate solubilization and acid production. When inoculated with PRGB06 in vitro and in gnotobiotic condition, red pepper and maize showed increase in root length, seedling vigor and dry bio-mass.

Construction of fat1 Gene Expression Vector and Its Catalysis Efficiency in Bovine Fetal Fibroblast Cells

  • Liu, Boyang;Yang, Runjun;Li, Junya;Zhang, Lupei;Liu, Jing;Lu, Chunyan;Lian, Chuanjiang;Li, Zezhong;Zhang, Yong-Hong;Zhang, Liying;Zhao, Zhihui
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.621-628
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    • 2012
  • The FAT-1 protein is an n-3 fatty acid desaturase, which can recognize a range of 18- and 20-carbon n-6 substrates and transform n-6 polyunsaturated fatty acids (PUFAs) into n-3 PUFAs while n-3 PUFAs have beneficial effect on human health. Fat1 gene is the coding sequence from Caenorhabditis elegans which might play an important role on lipometabolism. To reveal the function of fat1 gene in bovine fetal fibroblast cells and gain the best cell nuclear donor for transgenic bovines, the codon of fat1 sequence was optimized based on the codon usage frequency preference of bovine muscle protein, and directionally cloned into the eukaryotic expression vector pEF-GFP. After identifying by restrictive enzyme digests with AatII/XbaI and sequencing, the fusion plasmid pEF-GFP-fat1 was identified successfully. The pEF-GFP-fat1 vector was transfected into bovine fetal fibroblast cells mediated by Lipofectamine2000$^{TM}$. The positive bovine fetal fibroblast cells were selected by G418 and detected by RT-PCR. The results showed that a 1,234 bp transcription was amplified by reverse transcription PCR and the positive transgenic fat1 cell line was successfully established. Then the expression level of fat1 gene in positive cells was detected using quantitative PCR, and the catalysis efficiency was detected by gas chromatography. The results demonstrated that the catalysis efficiency of fat1 was significantly high, which can improve the total PUFAs rich in EPA, DHA and DPA. Construction and expression of pEF-GFP-fat1 vector should be helpful for further understanding the mechanism of regulation of fat1 in vitro. It could also be the first step in the production of fat1 transgenic cattle.

Effect of Corn Silage and Soybean Silage Mixture on Rumen Fermentation Characteristics In Vitro, and Growth Performance and Meat Grade of Hanwoo Steers (옥수수 사일리지와 대두 사일리지의 혼합급여가 In Vitro 반추위 발효성상 및 거세한우의 성장과 육질등급에 미치는 영향)

  • Kang, Juhui;Lee, Kihwan;Marbun, Tabita Dameria;Song, Jaeyong;Kwon, Chan Ho;Yoon, Duhak;Seo, Jin-Dong;Jo, Young Min;Kim, Jin Yeoul;Kim, Eun Joong
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.42 no.2
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    • pp.61-72
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    • 2022
  • The present study was conducted to examine the effect of soybean silage as a crude protein supplement for corn silage in the diet of Hanwoo steers. The first experiment was conducted to evaluate the effect of replacing corn silage with soybean silage at different levels on rumen fermentation characteristics in vitro. Commercially-purchased corn silage was replaced with 0, 4, 8, or 12% of soybean silage. Half gram of the substrate was added to 50 mL of buffer and rumen fluid from Hanwoo cows, and then incubated at 39℃ for 0, 3, 6, 12, 24, and 48 h. At 24 h, the pH of the control (corn silage only) was lower (p<0.05) than that of soybean-supplemented silages, and the pH numerically increased along with increasing proportions of soybean silage. Other rumen parameters, including gas production, ammonia nitrogen, and total volatile fatty acids, were variable. However, they tended to increase with increasing proportions of soybean silage. In the second experiment, 60 Hanwoo steers were allocated to one of three dietary treatments, namely, CON (concentrate with Italian ryegrass), CS (concentrate with corn silage), CS4% (concentrate with corn silage and 4% of soybean silage). Animals were offered experimental diets for 110 days during the growing period and then finished with typified beef diets that were commercially available to evaluate the effect of soybean silage on animal performance and meat quality. With the soybean silage, the weight gain and feed efficiency of the animal were more significant than those of the other treatments during the growing period (p<0.05). However, the dietary treatments had little effect on meat quality except for meat color. In conclusion, corn silage mixed with soybean silage even at a lower level provided a greater ruminal environment and animal performances, particularly with increased carcass weight and feed efficiency during growing period.

Effects of Mustard Seeds and Powder on In vitro Ruminal Fermentation Characteristics and Methane Production (겨자종자와 겨자분의 첨가가 반추위 발효성상과 메탄생성에 미치는 영향)

  • Lee, Kang Yeon;Kim, Kyoung Hoon;Baek, Youl Chang;Ok, Ji Un;Seol, Yong Joo;Han, Ki Jun;Park, Keun Kyu;Ryu, Ho Tae;Lee, Sang Suk;Jeon, Che Ok;Oh, Young Kyoon
    • Journal of Animal Science and Technology
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    • v.55 no.1
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    • pp.25-32
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    • 2013
  • The purpose of this study was to investigate the effects of mustard, which contains allyl isothiocyanate, on ruminal fermentation and methane emission in vitro. To this end, diluted ruminal fluid(30ml) was incubated anaerobically at $39^{\circ}C$ or 6, 12, and 24 h with or without seeds or powdered mustard. Either mustard seed or powdered mustard was weighed and serially (0, 3.33, 5.00, 6.67, and 8.33 g/L) mixed with ruminal fluid. Ammonia-N was increased (P < 0.05) by mustard treatment in a dose dependent manner. Regardless of concentration or form, mustard increased (P < 0.05) total VFA content but decreased (P < 0.01) pH compared to control group. Molar proportion of acetate (A) was decreased (P < 0.05) whereas propionate (P) was increased (P < 0.05) by mustard treatment, thereby A:P ratio was decreased (P < 0.05) compared to control group. Total gas production was increased (P < 0.01) in a linear manner by mustard treatment compared to control group. There was no effect of mustard powder, except 8.33 g/L level at 6 h, on methane emission. However, at 24 h, methane emission was reduced (P < 0.05) by 4.77% and 11.54% with 6.67 g/L and 8.33 g/L of mustard seeds supplementation, respectively. Altogether, these results suggest that mustard seeds containing allyl isothiocyanate may reduce methane production without disturbing ruminal fermentation.

Some Prophylactic Options to Mitigate Methane Emi ssion from Animal Agriculture in Japan

  • Takahashi, Junichi
    • 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.

Endophytic Trichoderma gamsii YIM PH30019: a promising biocontrol agent with hyperosmolar, mycoparasitism, and antagonistic activities of induced volatile organic compounds on root-rot pathogenic fungi of Panax notoginseng

  • Chen, Jin-Lian;Sun, Shi-Zhong;Miao, Cui-Ping;Wu, Kai;Chen, You-Wei;Xu, Li-Hua;Guan, Hui-Lin;Zhao, Li-Xing
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.315-324
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    • 2016
  • Background: Biocontrol agents are regarded as promising and environmental friendly approaches as agrochemicals for phytodiseases that cause serious environmental and health problems. Trichoderma species have been widely used in suppression of soil-borne pathogens. In this study, an endophytic fungus, Trichoderma gamsii YIM PH30019, from healthy Panax notoginseng root was investigated for its biocontrol potential. Methods: In vitro detached healthy roots, and pot and field experiments were used to investigate the pathogenicity and biocontrol efficacy of T. gamsii YIM PH30019 to the host plant. The antagonistic mechanisms against test phytopathogens were analyzed using dual culture, scanning electron microscopy, and volatile organic compounds (VOCs). Tolerance to chemical fertilizers was also tested in a series of concentrations. Results: The results indicated that T. gamsii YIM PH30019 was nonpathogenic to the host, presented appreciable biocontrol efficacy, and could tolerate chemical fertilizer concentrations of up to 20%. T. gamsii YIM PH30019 displayed antagonistic activities against the pathogenic fungi of P. notoginseng via production of VOCs. On the basis of gas chromatography-mass spectrometry, VOCs were identified as dimethyl disulfide, dibenzofuran, methanethiol, ketones, etc., which are effective ingredients for antagonistic activity. T. gamsii YIM PH30019 was able to improve the seedlings' emergence and protect P. notoginseng plants from soil-borne disease in the continuous cropping field tests. Conclusion: The results suggest that the endophytic fungus T. gamsii YIM PH30019 may have a good potential as a biological control agent against notoginseng phytodiseases and can provide a clue to further illuminate the interactions between Trichoderma and phytopathogens.

Production of $C_{21}$-Steroids in Longchin Goby, Chasmichthys dolichognathus during Oocyte Maturation (점망둑(Chasmichthys dolichognathus)의 성숙기 난모세포에서의 $C_{21}$-스테로이드 생성)

  • Baek, Hea-Ja
    • Development and Reproduction
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    • v.12 no.1
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    • pp.107-112
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    • 2008
  • To investigate the $C_{21}$-steroids produced from maturating oocytes in the longchin goby, Chasmichthys dolichognathus, the oocytes ($0.74{\sim}0.97\;mm$) were incubated with radiolabeled $17{\alpha}$-hydroxyprogesterone ($^3H-17{\alpha}OHP$) for 24 hours. The resulting metabolites were analyzed by thin layer chromatography and identified by gas chromatography-mass spectrometry. Two $C_{21}$-steroids, $17{\alpha}$-hydroxy, $20{\alpha}$-dihydroprogesterone ($17{\alpha}20{\alpha}P$) and $17{\alpha}$-hydroxy, $20{\beta}$-dihydroprogesterone ($17{\alpha}20{\beta}P$), were converted from $^3H-17{\alpha}OHP$ in the maturing oocytes. These two main metabolites were detected at 0.80 mm diameter oocytes or greater. In addition, the effects of these metabolites on in vitro germinal vesicle breakdown (GVBD) were tested. The sensitivity of oocytes to the induction of GVBD was greater at $17{\alpha}20{\beta}P$ than $17{\alpha}20{\alpha}P$. This result showed that $17{\alpha}20{\beta}P$ is a major maturation inducing steroid (MIS) in longchin goby, suggesting $17{\alpha}20{\alpha}P$ may play a role in regulating the oocyte maturation process.

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Analysis of the Component and Immunological Efficacy of Chamaecyparis obtusa Leaf Extract (편백나무 잎 추출물의 성분분석과 면역효능에 관한 연구)

  • Kim, Joung Hee;Lee, Syng-Ook;Do, Kook Bae;Ji, Won Dae;Kim, Sun Gun;Back, Young Doo;Kim, Keuk-Jun
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.1
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    • pp.37-43
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    • 2018
  • Chamaecyparis obtusa (CO) has recently been attracting attention because of its beneficial effects on skin allergies, atopic dermatitis, and skin diseases, such as acne and eczema. In the present study, the extract from CO leaf grown in Jangseong gun, Jeollanam-do, Korea was evaluated for its anti-oxidant, anti-inflammatory, and anti-allergic effects in vitro. The total polyphenol content of the CO leaf extract was $25.89{\pm}0.31mg$ gallic acid equivalents (GAE)/g. Gas-chromatography mass-spectrometry (GC-MS) analysis revealed the presence of six compounds in the CO leaf extract: ${\alpha}-terpinene$ (3.03 mg/g), ${\alpha}-terpineol$ (9.48 mg/g), limonene (5.96 mg/g), borneol (59.78 mg/g), myrcene (4.85 mg/g), and sabinene (11.31 mg/g). The $RC_{50}$ values of the CO leaf extract for $H_2O_2$ and ABTS radical were $5.47{\pm}0.13mg/mL$ and $4.00{\pm}0.01mg/mL$, respectively. In addition, the CO leaf extract showed significant inhibitory effects on lipopolysaccharide-induced nitric oxide production in RAW 264.7 cells and IgE-induced release of ${\beta}-hexosaminidase$ (degranulation) in mast-cell like RBL-2H3 cells. The cell viability assay showed that the CO leaf extract ($100{\sim}800{\mu}g/mL$) did not affect the viability of human normal skin fibroblast CCD-986sk cells significantly. Overall, these results suggest that the CO leaf extract is a potential functional cosmetic ingredient that can exert anti-oxidant, anti-inflammatory, and anti-allergic effects.