• Title/Summary/Keyword: Anaerobic culture

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Isolation of Anaerobic Cellulolytic Bacteria from the Rumen of Holstein Dairy Cows to Develop Feed Additives for Ruminants (반추동물용 사료첨가제개발을 위한 홀스타인 젖소의 반추위로부터 분리한 혐기성 섬유소 분해균의 특성연구)

  • Choi, Nag-Jin;Lee, Gi-Young;Jeong, Kwang-Hwa;Kim, Chang-Hyun
    • Korean Journal of Organic Agriculture
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    • v.20 no.3
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    • pp.327-343
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    • 2012
  • In order to develop a high cellulolytic direct-fed microorganism (DFM) for ruminant productivity improvement, this study isolated cellulolytic bacteria from the rumen of Holstein dairy cows, and compared their cellulolytic abilities via DM degradability, gas production and cellulolytic enzyme activities. Twenty six bacteria were isolated from colonies grown in Dehority's artificial (DA) medium with 2% agar and cultured in DA medium containing filter paper at $39^{\circ}C$ for 24h. 16s rDNA gene sequencing of four strains from isolated bacteria showed that H8, H20 and H25 strains identified as Ruminococcus flavefaciens, and H23 strain identified as Fibrobacter succinogenes. H20 strain had higher degradability of filter paper compared with others during the incubation. H8 (R. flavefaciens), H20 (R. flavefaciens), H23 (F. succinogenes), H25 (R. flavefaciens) and RF (R. flavefaciens sijpesteijn, ATCC 19208) were cultured in DA medium with filter paper as a single carbon source for 0, 1, 2, 3, 4 and 6 days without shaking at $39^{\circ}C$, respectively. Dry matter degradability rates of H20, H23 and H25 were relatively higher than those of H8 and RF since 2 d incubation. The cumulative gas production of isolated cellulolytic bacteria increased with incubation time. At every incubation time, the gas production was highest in H20 strain. The activities of carboxymethylcellulase (CMCase) and Avicelase in the culture supernatant were significantly higher in H20 strain compared with others at every incubation time (p<0.05). Therefore, although further researches are required, the present results suggest that H20 strain could be a candidate of DFM in animal feed due to high cellulolytic ability.

Effect on the Growth of Chinese Cabbage and Chemical Composition of the Liquid Piggery Slurry Digested Anaerobically in Biogas Plant (바이오가스 프랜트에서 혐기 소화시킨 돈사 Slurry의 화학적 특성 및 배추생육 시용효과에 관한 연구)

  • Jeong, Chan-Seong;Bak, Jeong-Geun;Roh, Ahn-Sung;Cho, Woong-Gi;Lee, Sung-Sill;Moon, Yea-Hwang
    • Journal of Animal Environmental Science
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    • v.20 no.3
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    • pp.125-132
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    • 2014
  • This study was conducted to investigate the effects on chemical properties of soil and yield of chinese cabbage when the piggery liquid slurry (ADS) digested anaerobically in the biogas plant was sprinkled to the soils. Treatments were composed of non-nitrogen plot (PK), chemical fertilizer plot (NPK) and ADS plot, and were arranged by randomized block design with 3 replications. The ADS was an alkali (pH 8.6) fertilizer containing 0.55% of total nitrogen, optimal levels of phosphorus, potassium and other components. When based (100%) on the yield of chinese cabbage in the NPK, those of the ADS and the PK were 114% and 43%, respectively. The nitrogen absorption of chinese cabbage at 50-d after transplant was significantly greater (p<0.05) in the ADS than other treatment. Heavy metal contents of soil were not difference between before and after experiment. With supplementing phosphorus and potassium sources in it, the ADS should be an excellent fertilizer in cabbage culture.

Prokaryotic Communities of Halophilic Methylotrophs Enriched from a Solar Saltern (염전으로부터 농화배양된 호염 메틸영양미생물 군집의 특성)

  • Kim, Jong-Geol;Park, Soo-Je;Rhee, Sung-Keun
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.286-290
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    • 2010
  • C-1 compounds are observed in anaerobic sediment of high salt environments. Thus, surface sediments and waters from these environments are therefore potential habitats for aerobic methylotrophic microorganisms. The soil samples collected from saltern and tidal flat as inoculums and methanol as carbon and energy source was supplied. After subculture depending on the salt concentration, methanol oxidizing bacteria growth condition investigated, the results of methanol oxidizing bacteria can grow in salt conditions, and the maximum concentration was 20%. Analysis based on denaturing gradient gel electrophoresis of 16S rRNA genes indicates that Methelyophaga-like bacteria were dominants of methylotrophs in the enrichment culture. Quantitative PCR showed that archaeal cells were about 1-10% of bacterial cells. Additionally archaea were assumed not to be involved in methanol oxidation since bacterial antibiotics completely blocked the methanol oxidation. Our results suggest that Methelyophaga-like bacteria could be involved in C-1 compounds oxidation in hypersaline environments although those activities are sensitive to salinity above 20%.

Effect of NADH-Dependent Enzymes Related to Oxygen Metabolism on Elimination of Oxygen-Stress of Bifidobacteria (NADH요구 산소대사관련 효소가 bifidobacteria의 산소스트레스 제거에 미치는 영향)

  • Ahn, Jun-Bae;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.37 no.6
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    • pp.951-956
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    • 2005
  • Selection of oxygen-tolerant strains and elucidation of their oxygen tolerance mechanism were crucial for effective use of bifidobacteria. Oxygen-tolerant bifidobacteria were able to significantly remove environmental oxygen (oxygen removal activity) as compared to oxygen-sensitive strains. Most oxygen removal activity was inhibited by heat treatment and exposure to extreme pH (2.0) of bifidobacterial cell. NADH oxidase was major enzyme related to oxygen removal activity. Oxygen-tolerant bifidobacteria possessed high NADH peroxidase activity level to detoxify $H_2O_2$ formed from reaction of NADH oxidase. Addition of oxygen to anaerobic culture broth significantly increased activities of HADH oxidase and NADH peroxidase within 1hr and rapid increment of oxygen concentration was prevented. Results showed NADH oxidase and NADH peroxidase of oxygen-tolerant bifidobacteria played important roles in elimination of oxygen and oxygen metabolite $(H_2O_2)$.

Continuous Nitrate Reduction by Gel and Foam Matrix (GFM) Immobilized Cells (겔-스폰지 혼용 시스템에 고정화된 세포를 통한 질산염의 연속적 분해)

  • Cho, Yang-Hee;Hahm, Tae-Shik
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.902-907
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    • 1998
  • In this study the anaerobic degradation of nitrate by in GFM (gel and foam matrix) and bead gel immobilized Paracoccus denitrificans DSM 65 in continous culture was conducted. A novel GFM immobilization system was developed in order to improve conventional system (bead). With increasing nitrate concentration in water, the nitrate reduction rate was increased. The observed maximum denitrification rate by in GFM immobilized cells was 177 mg/L h in buffered water, while that was 33 mg/L h in tap water. In comparison with bead system the reduction activity by GFM system showed $1.2{\sim}2.1$ times better. The denitrification activity was not changed after 16 days storage at $5^{\circ}C$ and also showed better activity than that of free cells or even bead immobilized cells.

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Effects of Dietary n-3/n-6 Fatty Acid Ratio on In Vitro Fermentation Characteristics and Fatty Acid Profiles

  • Kim, Dong-Hyeon;Amanullah, Sadar M.;Yoon, Hee;Lee, Hyuk-Jun;Kong, Il-Keun;Kim, Sam-Churl;Cho, Kyu-Woan;Kim, Sang-Bum
    • Journal of agriculture & life science
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    • v.46 no.3
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    • pp.79-85
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    • 2012
  • This study was conducted to examine the effect of dietary n-3/n-6 fatty acid (FA) ratio on in vitro dry matter digestibility (IVDMD), fermentation indices and FA profile. Rice bran was mixed with oil sources (cotton seed oil and linseed oil) to make the diets at 0.02, 0.29 and 0.61 of dietary n-3/n-6 FA ratio. These diets (0.5g) were placed into the incubation bottles with 40 ml of anaerobic culture medium, which contained rumen fluid and Van Soest medium at 1:2 ratio. Five replicates of each diet and two blanks were incubated at $39^{\circ}C$ for 48 hours. After incubation, the incubated contents were centrifuged. The residues were freeze-dried for DMD and FA analyses. The supernatant was used for pH, $NH_3-N$ and volatile fatty acid analyses. The concentrations of lactate (p<0.001) and iso-valerate (p<0.001) decreased linearly with increasing dietary n-3/n-6 FA ratio, but acetate concentration (p=0.056) and the ratio of acetate to propionate (p=0.005) was increased linearly. The concentrations of n-3, n-6 FA and the ratio of n-3/n-6 FA in residues increased (p<0.001) linearly with increasing dietary n-3/n-6 FA ratio, but C18:1n-9 FA concentration was decreased (p<0.001) linearly. With these results, it could affect fermentation characteristics and FA profile of rumen content by dietary n-3/n-6 FA ratio.

Study on the Effects of Supplemented Factors on the Production of Vitamin $B_{12}$ by Propionibacterium shermanii (Propionibacterium shermanii에 의한 비타민 $B_{12}$의 생성에 영향을 미치는 배지첨가물들에 대한 연구)

  • Kim, Ji-Young;Kim, Kong-Hwan;Kim, Kyoung-Ja;Goo, Yang-Mo
    • YAKHAK HOEJI
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    • v.38 no.5
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    • pp.614-620
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    • 1994
  • Following the study on the fermentation conditions influencing the production of vitamin $B_{12}$ by Propionibacterium shermanii(Korean J Biotechnol. Bioeng. 7,126-131, 1992), the effects of some factors supplemented in the medium on the production of vitamin $B_{12}$ were studied. Maximum production of vitamin $B_{12}$ was observed when $Co^{+2}$ was supplemented at the concentration of 2-4 ppm in the fermentation medium. Increase of the supplemented $Co^{+2}$ to 12 ppm did not inhibit the growth of the organism, but it accelerated the lysis of the organism. In the literature, peptone was reported to activate the biosynthesis of vitamin $B_{12}$. Examination of the effect of peptone on the growth and the production of vitamin $B_{12}$ showed that at early stage more vitamin$B_{12}$ was observed in the supplemented medium, but no difference was observed in the later stage of fermentation. Examination of the time for addition and the amount of 5,6-dimethylbenzimidazole, a precursor known to influence the production of vitamin $B_{12}$, showed that a maximum yield of vitamin $B_{12}$ was observed when 15 mg/L was added to the fermentation medium after 2 days' incubation. The effect was comparable with the increase of the production of vitamin $B_{12}$ when the fermentation condition was changed to aerobic condition after 2 days' culture under anaerobic condition.

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Dual Culture Inoculation Enhanced Quality of Silage Produced from Leguminous Plants

  • Muthusamy, Karnan;Ilavenil, Soundharrajan;Jung, Jeong Sung;Lee, Bae Hun;Park, Hyung Soo;Choi, Ki Choon
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.41 no.3
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    • pp.176-182
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    • 2021
  • Ensiling is the most preferred technology to preserve the silage quality with high nutrients by the presence of lactic acid bacteria. In this study, lactic acid bacteria RJ1 and S22 were used to make the silages from different leguminous plants such as alfalfa, hairy vetch and red clover. Experimental groups were divided into control and LAB inoculated groups. LAB inoculated group; all legumes treated with a mixture of RJ1 and S22 and made an anaerobic condition for 45d. Without the addition of LAB considered the control group. The results showed that the lactic acid content was higher in all silages in response to LAB treatment and acetic acid content was slightly increased except red clover by LAB compared to control silages. A poor silage quality marker butyric acid was reduced all legume silages in response to LAB inoculation than control silages. The organic acid is closely associated with microbial population experimental silages. We noted that higher LAB and lower yeast were found in the silage in response to LAB treatment. The contents of crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), and total digestible nutrient (TDN) were not altered significantly between control and LAB treated silages. Overall data suggested that the inclusion of additional LAB potentially enhance the silage quality and preserved the nutrients for long period.

The effect of rosehip extract on TNF-α, IL-1β, and IL-8 production in THP-1-derived macrophages infected with Aggregatibacter actinomycetemcomitans

  • Song, Yuri;Kim, Si young;Chung, Jin
    • International Journal of Oral Biology
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    • v.47 no.1
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    • pp.1-8
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    • 2022
  • Inflammation is a protective mechanism against pathogens, but if maintained continuously, it destroys tissue structures. Aggregatibacter actinomycetemcomitans is a gram-negative, facultative anaerobic bacterium often found in severe periodontitis. A. actinomycetemcomitans invades epithelial cells and triggers inflammatory response in the immune cells. In this study, we investigated the effect of water-soluble rosehip extract on A. actinomycetemcomitans-induced inflammatory responses. A human monocytic cell line (THP-1) was differentiated to macrophages by phorbol 12-mystristate 13-acetate treatment. The cytotoxic effect of extract was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The effects of extract on bacterial growth were examined by measuring the optical densities using a spectrophotometer. THP-1-derived macrophages were infected A. actinomycetemcomitans after extract treatment, and culture supernatants were analyzed for cytokine production using enzyme-linked immunosorbent assay. Protein expression was measured by western blotting. Extract was not toxic to THP-1-derived macrophages. A. actinomycetemcomitans growth was inhibited by 1% extract. The extract suppressed A. actinomycetemcomitans-induced tumor necrosis factor-α, interleukin (IL)-1β, and IL-8 production. It also decreased mitogen-activated protein kinase (MAP kinase) and nuclear factor-κB (NF-κB) phosphorylation. Moreover, the extract inhibited the expression of inflammasome components, including nucleotide-binding oligomerization domain-like receptor pyrin domain-containing protein 3, Absent in Melanoma 2, and apoptosis associated speck-like protein containing a CARD. And cysteine-aspartic proteases-1 and IL-1β expression were decreased by the extract. In summary, extract suppressed A. actinomycetemcomitans growth and decreased inflammatory cytokine production by inhibiting activation of MAP kinase, NF-κB, and inflammasome signaling. Rosehip extract could be effective in the treatment of periodontal inflammation induced by A. actinomycetemcomitans infection.

Boosting Power Generation by Sediment Microbial Fuel Cell in Oil-Contaminated Sediment Amended with Gasoline/Kerosene

  • Aleman-Gama, Elizabeth;Cornejo-Martell, Alan J.;Kamaraj, Sathish Kumar;Juarez, Katy;Silva-Martinez, Susana;Alvarez-Gallegos, Alberto
    • Journal of Electrochemical Science and Technology
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    • v.13 no.2
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    • pp.308-320
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    • 2022
  • The high internal resistance (Rint) that develops across the sediment microbial fuel cells (SMFC) limits their power production (~4/10 mW m-2) that can be recovered from an initial oil-contaminated sediment (OCS). In the anolyte, Rint is related to poor biodegradation activity, quality and quantity of contaminant content in the sediment and anode material. While on the catholyte, Rint depends on the properties of the catholyte, the oxygen reduction reaction (ORR), and the cathode material. In this work, the main factors limiting the power output of the SMFC have been minimized. The power output of the SMFC was increased (47 times from its initial value, ~4 mW m-2) minimizing the SMFC Rint (28 times from its initial value, 5000 ohms), following the main modifications. Anolyte: the initial OCS was amended with several amounts of gasoline and kerosene. The best anaerobic microbial activity of indigenous populations was better adapted (without more culture media) to 3 g of kerosene. Catholyte: ORR was catalyzed in birnessite/carbon fabric (CF)-cathode at pH 2, 0.8M Na2SO4. At the class level, the main microbial groups (Gammaproteobacteria, Coriobacteriia, Actinobacteria, Alphaproteobacteria) with electroactive members were found at C-anode and were associated with the high-power densities obtained. Gasoline is more difficult to biodegrade than kerosene. However, in both cases, SMFC biodegradation activity and power output are increased when ORR is performed on birnessite/CF in 0.8 M Na2SO4 at pH 2. The work discussed here can focus on bioremediation (in heavy OCS) or energy production in future work.