Anaerobic ammonium oxidation (AMX) is a cost-efficient biological nitrogen removal process. The coexistence of ammonia-oxidizing bacteria (AOB) in an AMX reactor is an interesting research topic as a nitrogen-related bacterial consortium. In this study, a sequencing batch reactor for AMX (AMX-SBR) was operated with a conventional activated sludge. The AOB in an AMX bioreactor were identified and quantified using terminal restriction fragment length polymorphism (T-RFLP) and real-time qPCR. A T-RFLP assay based on the ammonia monooxygenase subunit A (amoA) gene sequences showed the presence of Nitrosomonas europaea-like AOB in the AMX-SBR. A phylogenetic tree based on the sequenced amoA gene showed that AOB were affiliated with the Nitrosomonas europaea/mobilis cluster. Throughout the enrichment period, the AOB population was stable with predominant Nitrosomonas europaea-like AOB. Two OTUs of amoA_SBR_JJY_20 (FJ577843) and amoA_SBR_JJY_9 (FJ577849) are similar to the clones from AMX-related environments. Real-time qPCR was used to quantify AOB populations over time. Interestingly, the exponential growth of AOB populations was observed during the substrate inhibition of the AMX bacteria. The specific growth rate of AOB under anaerobic conditions was only 0.111 d-1. The growth property of Nitrosomonas europaea-like AOB may provide fundamental information about the metabolic relationship between the AMX bacteria and AOB.
In recent, multi-sector nutriculture in which each sectors are managed with different compositions and supplying amounts of nutrient solution is being prevalent especially in large-scale greenhouses. This study was carried out to develop an automatic mixing and supplying system of nutrient solution for the multi-sector greenhouse which can control various compositions of nutrient solution and supplying amounts for each sector. The stock solution control device could control discharge of the stock solution with device actuating time. The experimental result showed high correlation between the discharge and the actuating time. The system could make nutrient solution of various compositions and showed possibility of changing compositions of nutrient solution according to changes of growth stages or environments. Undesirable remnant nutrient-solution effect on the next supply was decreased by lowering RC of the previous nutrient solution remaining in the tank and supply pipe through rinse process. To evaluate performance of the developed system, experiments that compare N $O_3$$^{[-10]}$ , and P $O_4$$^{3-}$ concentrations of nutrient solution made in laboratory level with those made by the developed system were conducted. The experimental results showed that the deviations for N $O_3$$^{[-10]}$ , and P $O_4$$^{3-}$ concentrations of nutrient solution composited by the developed system were -2.36% and -5.15%, respectively.
To evaluate the effects of microorganism agents on oil biodegradation, treatability and microcosm studies were conducted. Petroleum oil degrading bacteria were isolated from enriched cultures of oil-contaminated sediment samples using a mineral salts medium (MSM) containing 0.5% Arabian heavy crude oil as the sole carbon source. After a 5 day-incubation period using MSM, mixed microorganisms of three species (strains BS1, BS2 and BS4) degraded 48.4% of aliphatic hydrocarbons and 30.5% of aromatic hydrocarbons. Treatability and microcosm tests were performed in the three different treatment conditions (AO: Arabian heavy crude oil, AO+IN: Arabian heavy crude oil+inorganic nutrient, AO+IN+MM: Arabian heavy crude oil+inorganic nutrient+mixed microorganism agents). Among these, significantly enhanced biodegradation of aliphatic hydrocarbons were observed in AO+IN and AO+IN+MM conditions, without showing any different biodegradation rates in either condition. However, the degradation rates of aromatic hydrocarbons in an AO+IN+MM condition were increased by 50% in the treatability test and by 13% in the microcosm test compared to those in an AO+IN condition. Taken together, it can be concluded that mixed microorganism agents enhance the biodegradation of aliphatic and aromatic hydrocarbons in laboratory, a treatability test, and a microcosm test. This agent could especially be a useful tool in the application of bioremediation for removal of aromatic hydrocarbons.
This experiment was carried out to examine the effects of culture medium on cell growth of the viola (Viola patrinii DC.) suspension culture. The results are as follows: The greatest cell growth rates were found with MS medium suggesting that this medium could be recommendable for the viola suspension cell culture. When the nitrogen sources (NH$_4$NO$_3$ and KNO$_3$) of MS salts were diluted at half concentrations, the cell growth rates were slightly increased, but when the combined concentration rations of NH$_4$+ and NO$_3$ions were 25 to 75 the greatest cell growth rates were obtained. This result imply that the nitrogen ion sources had slight influence on the rates. Another feature was obtained. This result implys that the nitrogen ion sources had slight influence on the rates. Another feature was that as the concentration of NH$_4$+ ion lowered, the callus color changed to pale yellow with some red spots. The addition of casein hydrolysate (5 g/L) was more effective for the cell growth. On the basis of microscopic observation, the highest cell growth rates were detected during 2-4 weeks culture and after 6 weeks of the culture, some elongated and vacuolated cells were determined.
Park, Joong-Kook;Lim, Dong-Hyun;Kim, Sang-Bum;Ki, Kwang-Seok;Lee, Hyun-June;Kwon, Eung-Gi;Cho, Won-Mo;Kim, Chang-Hyun
Journal of Animal Science and Technology
/
v.53
no.5
/
pp.441-450
/
2011
This study was conducted to determine the effects of partial replacement of corn grain and soybean meal with agricultural by-product feeds on in vitro rumen fermentation characteristics and optimum levels of mixing ratio. The agricultural by-products to examine the effectiveness of the partial replacement of concentrate were wheat bran, corn gluten feed, bakery waste, soybean curd, rice bran, green kernel rice, soybean hull, distillers' grain, and mushroom substrate. In the first experiment, in vitro ruminal fermentation characteristics of feedstuffs were evaluated at 0, 3, 6, 12, 24, and 48 hours after incubation. In the second experiment, fermentation characteristics were investigated with green kernel rice and soybean curd which replaced corn grain or soybean meal. Feed were formulated with 40% corn grain + 20% soybean meal (T1), 40% corn grain + 17.5% soybean meal + 2.5% soybean curd (T2), 25% corn grain + 20% soybean meal + 15% green kernel rice (T3), and 30% corn grain + 15% soybean meal + 6% green kernel rice + 9% soybean curd (T4), respectively, with forage source of 10% alfalfa hay, 20% timothy hay, and 10% corn silage as fed-basis. In 24 and 48 hour cultivations, T4 showed significantly lower pH compared to T1, whereas in 3 and 24 hour cultivations, T4 showed significantly higher DM degradation compared to T1. In addition, the gas production of T3 was also higher than T1 (p<0.05). Overall results of the present experiments indicated that green kernel rice and soybean curd as agricultural by-products have the possibility of partial replacements of corn grain and soybean meal.
These study were conducted to determine the effects of a whole or steam-flaked corn based diet on rumen microbial fermentation in vitro and ruminal metabolism in the Korean Native Goat(KNG) in vivo. The experiments consisted of two dietary treatments: control, steam-flaked corn(SFC) based diet(80%) + rice straw mixed(20%)(SFCR); 100% whole corn based diet(WC). The first experiment was conducted to investigate the effect of whole corn on ruminal metabolism in vitro for 0 to 48 h. pH values were optimally maintained during incubation time, and were not significantly different between treatments. Gas production of SFCR was significantly higher than WC(p<0.01). $NH_3$-N concentration tended to increase for WC, but not significantly different between treatments. The mean value of total volatile fatty acid concentration of WC was significantly lower than SFCR(p<0.01), but SFCR and WC linearly increased as the time of incubation approached 48 h. Mean value of acetate concentration of SFCR was significantly higher than WC(p<0.01). Propionate concentration of WC for the total incubation time was significantly higher than SFCR(p<0.01). The digestibility of dry matter was not significantly different between treatments, but SFCR was somewhat higher than WC. The second experiment was conducted to effect of whole shelled corn based diet on rumen metabolism in KNG. pH values tended to decrease through all treatments. There was not a significantly difference between treatments. Microbial protein yield of SFCR was significantly higher than WC(p<0.01). $NH_3$-N concentration of WC was significantly (p<0.01) higher than SFCR. Total VFA and propionate concentration of WC was significantly higher than SFCR(p<0.01), but acetate concentrate of WC was not significantly higher than SFCR. The mean value of total lactate concentration was significantly(p<0.01) different but the value of SFCR and WC were lower than the average concentration of acidosis. In sacco DM disappearance rate of SFC was significantly(p<0.01) higher than WC.
This study was conducted to evaluate the effects of saponin contained plant extracts on in vitro rumen fermentation characteristics and methane production. Ruminal fluid was collected from rumen cannulated Hanwoo steers fed rice straw and concentrate (5:5). Collected rumen fluids, corn starch and buffer including saponin contained plant extracts (ginseng, Ogapi, soapwort, tea plant and yucca; 0.5%/15 ml) were incubated at $39^{\circ}C$ for 24 h. All incubations were repeated five times. Rumen pH in all treatments was lower (p<0.05) compared with that of the control (no addition) during incubation time. The concentration of total VFA in all treatments was higher (p<0.05) than that of the control after 12h incubation. Compared with the control, the concentration of acetate and propionate in all treatments was lower and higher after 6h incubation, respectively. The concentration of $NH_3$-N in all treatments was lower (p<0.05) than that of the control except for Ogapi or yucca extracts supplementation. The number of protozoa in all treatments was significantly (p<0.05) lower than that of the control except for soapwort extract supplementation. The total gas production and methane production in all treatments was higher (p<0.05) and lower (p<0.05) compared with the control, except for ogapi or soapwort extracts supplementation after 12h incubation, respectively. Therefore, reduction in methane production by saponins may could be results from decreased protozoal population without any negative in vitro fermentation.
Kim, Sam-Soon;Lee, Ji-Yul;Park, Sung-Oh;Kim, Ki-Joo
The Korean Journal of Mycology
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v.1
no.2
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pp.15-23
/
1973
Mold producing cellulase were isolated from rotten woods, and identified as the three species: Aspergillus niger van Tieghem, Aspergillus schiemanni Thom and Trichoderma viride Pers. In this paper, culture conditions in the media and characteristics of these strains were investigated. Using these strains, we have conducted a research concerning the utilization of farm product waste materia's. 1. Optimum conditions for the cellulase formation were as follows. KM 10-1; pH 5.2-5.5, $35^{\circ}C$, incubation time 6 days. OL 11-1; pH 5.5, $30-35^{\circ}C$, incubation time 6 days. SH 9-2; pH 5.5, $30^{\circ}C$, incubatoin time 6 days. 2. Their cellulase activities in their optimum condition were as follows: KM 10-1; CMC-LP 78.5% CMC-SP 4.0 glucose mg/gm of the cultures/min. OL 11-1; CMC-LP 89.9%, CMC-SP 4.9 glucose mg/gm of the cultures/min. SH 9-2; CMC-I.P 77.4%, CMC-SP 3.9 glucose mg/gm of the cultures/min. 3. Hydrolysis of animal feed containing a large quantity (23-30%) of cellulose by means of the crude enzyme in the selected strains resolved 30% of the cellulose contained in the animal feed.
Journal of The Korean Society of Grassland and Forage Science
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v.41
no.2
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pp.119-127
/
2021
The objective of this study was to evaluate effects of feeding methods on in vitro ruminal fermentation, total gas and methane production in Hanwoo steers. Six Hanwoo steers fitted with rumen cannula (430 ± 21 kg of body weight) were randomly assigned to one of three feeding systems: 1) feeding forage 1 hour after concentrate, 2) feeding concentrate 1 hour after forage, 3) feeding mixed ration. Rumen fluid sampled from each animals was incubated 24 hours with maize or timothy substrates in in vitro. Ruminal pH was increased in feeding method 2 or maize substrate than that of other methods or timothy substrate (P < 0.001). The production of total volatile fatty acid, acetate, propionate, butyrate, and valerate were increased when steers fed diets using feeding method 1 or rumen fluid was incubated with maize substrate (P < 0.001). Increased production of total gas and methane was observed in feeding method 1 and maize substrate compared to those of other methods or timothy substrate (P < 0.001). Due to the inconsistent results between ruminal fermentation and gas production in this study, further research is required to estimate effects of feeding method on enteric fermentation and gas production in in vivo.
This study was conducted to investigate effects of terpenoids-rich plant extracts (TRPE) on the in vitro ruminal fermentation characteristics and methane production. The ruminal fluid was collected from a cannulated Hanwoo cow fed concentrate and timothy in the ratio of 6 to 4. The TRPE as Mint (Mentha arvensis var. piperascens), Pine (Pinus densiflora), Japan cedar (Cryptomeria japonica), Sichuan pepper (Zanthoxylum piperitum), Hinoki cypress (Chamaecyparis obtuse) and Japanese black pine (Pinus thunbergii) were used in this study. The 15 mL of mixture, contains McDougall buffer and rumen fluid in the ratio of 2 to 1. The mixture was dispensed anaerobically 50 mL serum bottles and it is contained 0.3 g timothy substrate and 5% TRPE. The bottles were incubated at $39^{\circ}C$ for 3, 6, 9, 12, 24, 48 and 72 hours. The pH value decrease by increased incubation times and the pH values at all times were significantly (p<0.05) higher in treatments than in control. The digestibility of dry matter at 3 hours was significantly (p<0.05) higher in mint treatment than in control. Productions of total gas and carbon dioxide at before 12 hours was significantly lower (p<0.05) in treatments than in control. The methane production at 24 hours was significantly (p<0.05) lower in treatments than in control. The concentrations of acetic acid and propionic acid at 24 hours were significantly higher (p<0.05) in mint and pine treatments than in control. In conclusion, the terpenoid-rich plant extracts were shown to decreased methane emission and without adversely affected ruminal fermentation. Therefore, the terpenoid-rich plant extracts as mint and pine were shown to decreased methane production and it has potential possibility for ruminal fermentations.
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