• Journal of Animal Science and Technology
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• v.65 no.2
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• pp.387-400
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• 2023

Ruminal protozoa, especially entodiniomorphs, engulf other members of the rumen microbiome in large numbers; and they release oligopeptides and amino acids, which can be fermented to ammonia and volatile fatty acids (VFAs) by amino acid-fermenting bacteria (AAFB). Studies using defaunated (protozoa-free) sheep have demonstrated that ruminal protozoa considerably increase intraruminal nitrogen recycling but decrease nitrogen utilization efficiency in ruminants. However, direct interactions between ruminal protozoa and AAFB have not been demonstrated because of their inability to establish axenic cultures of any ruminal protozoan. Thus, this study was performed to evaluate the interaction between Entodinium caudatum, which is the most predominant rumen ciliate species, and an AAFB consortium in terms of feed degradation and ammonia production along with the microbial population shift of select bacterial species (Prevotella ruminicola, Clostridium aminophilum, and Peptostreptococcus anaerobius). From an Ent. caudatum culture that had been maintained by daily feeding and transfers every 3 or 4 days, the bacteria and methanogens loosely associated with Ent. caudatum cells were removed by filtration and washing. An AAFB consortium was established by repeated transfers and enrichment with casamino acids as the sole substrate. The cultures of Ent. caudatum alone (Ec) and AAFB alone (AAFB) and the co-culture of Ent. caudatum and AAFB (Ec + AAFB) were set up in three replicates and incubated at 39℃ for 72 h. The digestibility of dry matter (DM) and fiber (NDF), VFA profiles, ammonia concentrations, pH, and microscopic counts of Ent. caudatum were compared among the three cultures. The co-culture of AAFB and Ent. caudatum enhanced DM degradation, VFA production, and Ent. caudatum cell counts; conversely, it decreased acetate: propionate ratio although the total bacterial abundance was similar between Ec and the Ec + AAFB co-culture after 24 h incubation. The ammonia production and relative abundance of C. aminophilum and P. anaerobius did not differ between AAFB alone and the Ec + AAFB co-culture. Our results indicate that Ent. caudatum and AAFB could have a mutualistic interaction that benefited each other, but their interactions were complex and might not increase ammoniagenesis. Further research should examine how such interactions affect the population dynamics of AAFB.

• Journal of Microbiology and Biotechnology
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• v.11 no.4
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• pp.700-703
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• 2001

The influence of process conditions on highly branched glucooligosaccharides production by mixed culture of Leuconostoc mesenteroides ATCC 13146 and Lipomyces starkeyi ATCC 74054 was studied. We divided the batch culture fermentations into two groups according to inoculation method. One-point inoculation was performed by coinoculation of L. mesenteroides and L. starkeyi at the ration of 10 to 1, and two-point inoculation by L. mesenteroides inoculation first and L. starkeyi inoculation after L. mesenteroides grew to the end of the log phase of growth. Two-point inoculation improved the yield of oligosaccharide by 1.5 to 20 fold more than one-point inoculation. In this process, the highest yield of oligosaccharides (48% of theoretical yield) and productivity (0.85 g/l/h) were obtained with starch as an initial substrate for L. starkeyi growth. The estimated composition of the end product consisted of 31.5% oligosaccharides, 17.6% dextran, and 46.5% mannitol.

• Journal of Microbiology and Biotechnology
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• v.3 no.2
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• pp.115-122
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• 1993

Poly(3-hydroxybutyrate)(P(3HB)) accumulation under nutrient-rich condition with various amounts of $(NH_4)_2 SO_4$ was systematically investigated. The results of the electron-microscopy and the solvent extraction showed that the P(3HB) accumulation is unavoidable even under nutrient-rich condition. This indicates that in a two-step culture of Alcaligenes eutrophus H16, the researches should be careful in interpreting the data of polyhydroxyalkanoates(PHAs) accumulation in terms of the carbon-source fed in the second step because the two-step culture product contains the P(3HB) produced under nutrient-rich condition. The polyester production capability in a two-step batch culture of A. eutrophus H16(ATCC 17699) was also investigated using various saccharides and their derivatives such as glucose, fructose, gluconic acid, glucaric acid, sorbitol, lactose, galactose, and mannose. The polyesters synthesized were characterized by 500 MHz$^{1}H-NMR$ spectroscopy, intrinsic viscosity$[\eta]$ measurement in chloroform and differential scanning calorimetry(DSC). 500 MHz $^{1}H-NMR$ analysis showed that all polyesters synthesized generally contained 1~2 mol% of 3HV. Another finding is that the glucose utilization can be increased by changing the autoclaving procedure of the substrate to enhance the P(3HB) production yield up to 46 wt% of P(3HB) in dry cells.

• ALGAE
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• v.28 no.1
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• pp.101-109
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• 2013

Some Chlorella species grow heterotrophically with organic substrate in dark condition. However, heterotrophic Chlorella species are limited and their optimum culture conditions are not fully known. In this study, three heterotrophic Chlorella species, two strains (C4-3 and C4-4) of C. vulgaris and one Chlorella sp. (C4-8) were examined on optimum culture conditions such as carbon source, temperature, and concentrations of nitrogen and phosphorus in Jaworski's medium (JM). And the growth and fatty acid composition of Chlorella were analyzed. For three heterotrophic Chlorella species, glucose (1-2%) as a carbon source only increased the growth and the range of optimum culture temperature was $26-28^{\circ}C$. Doubled concentrations of the nitrogen or phosphorus in JM medium also improved the growth of Chlorella. Chlorella cultured heterotrophically showed significantly higher growth rate and bigger cell size than those autotrophically did. C. vulgaris (C4-3) cultured heterotrophically showed the highest biomass in dry weight ($0.8g\;L^{-1}$) among three species. With respect to fatty acid composition, the contents of C16:0 and n-3 highly unsaturated fatty acid (HUFA) were significantly higher in autotrophic Chlorella than in heterotrophic one and those of total lipid were not different between different concentrations of nitrogen and phosphorus in JM medium. Among three Chlorella species in this study, C. vulgaris (C4-3) appeared to be the most ideal heterotrophic Chlorella species for industrial application since it had a high biomass and lipid content.

• Journal of Microbiology
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• v.44 no.1
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• pp.23-28
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• 2006

We investigated the effects of halogenated aromatic compounds (HACs) including naturally occurring ones (L-thyroxine, 3-chloro-L-tyrosine, 5-chloroindole, 2-chlorophenol, 4-chlorophenol and chlorobenzene) on polychlorinated biphenyl (PCB) dechlorination in sediment cultures. A PCB-dechlorinating enrichment culture of sediment microorganisms from the St. Lawrence River was used as an initial inoculum. When the culture was inoculated into Aroclor 1248 sediments amended with each of the six HACs, the extent of dechlorination was not enhanced by amendment with HACs. The dechlorination patterns in the HAC-amended sediments were nearly identical to that of the HAC-free sediments except the 3-chloro-L-tyrosine-amended ones where no dechlorination activity was observed. When these sediment cultures were transferred into fresh sediments with the same HACs, the dechlorination specificities remained the same as those of the initial inoculations. Thus, in the present study, the substrate range of the highly selected enrichment culture could not be broadened by the HACs. It appears that HACs affect PCB dechlorination mainly through population selection rather than enzyme induction of single population.

• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1928-1935
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• 2015

Microbial growth kinetics is often used to optimize environmental processes owing to its relation to the breakdown of substrate (contaminants). However, the quantification of bacterial populations in the environment is difficult owing to the challenges of monitoring a specific bacterial population within a diverse microbial community. Conventional methods are unable to detect and quantify the growth of individual strains separately in the mixed culture reactor. This work describes a novel quantitative PCR (qPCR)-based genomic approach to quantify each species in mixed culture and interpret its growth kinetics in the mixed system. Batch experiments were performed for both single and dual cultures of Pseudomonas putida and Escherichia coli K12 to obtain Monod kinetic parameters (μ_max and K_s). The growth curves and kinetics obtained by conventional methods (i.e., dry weight measurement and absorbance reading) were compared with that obtained by qPCR assay. We anticipate that the adoption of this qPCR-based genomic assay can contribute significantly to traditional microbial kinetics, modeling practice, and the operation of bioreactors, where handling of complex mixed cultures is required.

• Journal of Microbiology and Biotechnology
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• v.17 no.7
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• pp.1113-1119
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• 2007

Human bone marrow-derived mesenchymal stem cells (hBMMSCs) must differentiate into osteogenic cells to allow for successful bone regeneration. In this study, we investigated the effects of different combinations of three soluble osteogenic differentiation-inducing factors [L-ascorbic acid (AC), ${\beta}$-glycerophosphate (${\beta}G$), and bone morphogenic protein-2 (BMP-2)] and the presence of a hydroxyapatite (HA) substrate on hBMMSC osteogenic differentiation in vitro. hBMMSCs were cultured in medium containing various combinations of the soluble factors on culture plates with or without HA coating. After 7 days of culture, alkaline phosphatase (ALP) activity, calcium deposition, and osteoprotegerin (OPG) and osteopontin (OPN) expression were measured. The effects of individual and combined factors were evaluated using a factorial analysis method. BMP-2 predominantly affected expression of early markers of osteogenic differentiation (ALP and OPG). HA had the highest positive effect on OPN expression and calcium deposition. The interaction between AC, ${\beta}G$, and HA had the second highest positive effect on ALP activity.

• Journal of Korean Society of Water and Wastewater
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• v.8 no.1
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• pp.27-33
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• 1994

With the rapid industrialization, an ever-increasing quantity and kind of new organic compounds pose environmental problems due to their toxicity and physiological effect. However, research on the biodegradation of these compounds under anaerobic condition is very limited inspite of its efficiency and economical advantage. In this research, the pH effect on the ring cleavage of phenol under anaerobic condition was investigated, and the theory of phase separation was applied to the degradation of phenol for investigating the role of acidogenic bacteria. Results, obtained from biochemical methane potential(BMP) assay for 15.5 days of incubation, showed that acidic condition was more desirable for phenol degradation than alkaline condition. By both unacclimated methanogenic granular sludge and two mixed cultures, phenol was completely removed within six weeks of incubation with a gas conversion rate of over 86% of theoretical one. However, phenol was not degraded by unacclimated acidogenic culture, and thus it is considered as a syntrophic substrate. In case of phase separated biochemical methane potential(PSBMP) assay, in which acidogenic and methanogenic culture were seeded separately and consecutively, those that had been subjected to normal acidogens for 3 to 4 weeks showed higher gas production than those seeded with sterile or frozen culture.

• Journal of Microbiology
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• v.43 no.3
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• pp.301-307
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• 2005

The feasibility of laccase production by immobilization of Pleurotus ostreatus 1804 on polyurethane foam (PUF) cubes with respect to media composition was studied in both batch and reactor systems. Enhanced laccase yield was evidenced due to immobilization. A relatively high maximum laccase activity of 312.6 U was observed with immobilized mycelia in shake flasks compared to the maximum laccase activity of free mycelia (272.2 U). It is evident from this study that the culture conditions studied, i.e. biomass level, pH, substrate concentration, yeast extract concentration, $Cu^{2+}$ concentration, and alcohol nature, showed significant influence on the laccase yield. Gel electrophoretic analysis showed the molecular weight of the laccase produced by immobilized P. ostreatus to be 66 kDa. The laccase yield was significantly higher and more rapid in the packed bed reactor than in the shake flask experiments. A maximum laccase yield of 392.9 U was observed within 144 h of the fermentation period with complete glucose depletion.

• Journal of Aquaculture
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• v.10 no.2
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• pp.97-103
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• 1997

Larval growth of the oyster, Crassostrea nippona, was investigated at the different conditions such as water volume, depth of culture vessel and initiative feeding time at $23^{\circ}C$. As a result, morphological changes were occurred during the early larval growth. That is, shell length was higher than shell height at the time of D-shaped larvae. Later, the length became equal to the hight when their shell length reached $127\mum$and then shell height increased rapidly. Some of pediveliger larvae with the size of $360\mum\;in\;shell\;length\;and\;385\mum$ in shell height began to temporarily settle on the substrate after 18 days. The result also indicated that the depth of culture vessel turned out to be an important factor rather than water volume for the larval growth. The fist feeding time of oyster larvae appeared to be the first day after fertilization.