Tn order to be helpful to control dispersed microorganisms for stabLlization of wastewater treatment in a paper-makillg process, dominant strains were isolated aerobically and anaerobically. and identified and physiological characteristics were also analyzed. Pseudomonas carboxydohydrogena, Cardiobacten'm hominis, lvIicrococcus lylae, XanfomonCls campestris p" juglandis, Micrococcus diversus, and Comamonas terrigencl as aerobic dominants, and Streptococcus bovis and Prevotella buccae as anaerobIc dominants were identified fi'om the supernatent of the primary settling tank. It seemed that microflora in the treatment process would consist of many kinds of microorganisms, whose dominant would change easily according to environmental conditions, They all grew well at $37^{\circ}C$ and at different initial medium pH's. Especially, some of them required sulfate ion for their growth, which came from a chemical coagulant of aluminium sulfate in the primary settling tank. Interestingly. many anaerobes grew well even in the aerobic wastewater treatment process and seemed to have some functions. Population of anaerobes increased three times in the supematant of primary settling tank and ten times in Lhe bottom sludge of primary settling tank than in the prime wastewater. Therefore, these anaerobes contributed to the producH tion of offensive gases, which would make some microorganisms not precLpitate and be buoyant.
Ye, Gui-Zi;Jiang, Min;Li, Jian;Chen, Ke-Quan;Xi, Yong-Lan;Liu, Shu-Wen;Wei, Ping;Ouyang, Ping-Kai
Journal of Microbiology and Biotechnology
/
v.20
no.8
/
pp.1219-1225
/
2010
Actinobacillus succinogenes, a representative succinicacid-producing microorganism, is seriously inhibited by ammonium ions, thereby hampering the industrial use of A. succinogenes with ammonium-ion-based materials as the pH controller. Therefore, this study isolated an ammonium-ion-tolerant mutant of A. succinogenes using a continuous-culture technique in which all the environmental factors, besides the stress (ammonium ions), were kept constant. Instead of operating the mutant-generating system as a nutrient-limited chemostat, it was used as a nutrient-unlimited system, allowing the cells to be continuously cultured at the maximum specific growth rate. The mutants were isolated on agar plates containing the acid-base indicator bromothymol blue and a high level of ammonium ions that would normally kill the parent strain by 100%. When cultured in anaerobic bottles with an ammonium ion concentration of 354 mmol/l, the mutant YZ0819 produced 40.21 g/l of succinic acid with a yield of 80.4%, whereas the parent strain NJ113 was unable to grow. When using $NH_4OH$ to buffer the culture pH in a 3.0 l stirredbioreactor, YZ0819 produced 35.15 g/l of succinic acid with a yield of 70.3%, which was 155% higher than that produced by NJ113. In addition, the morphology of YZ0819 changed in the fermentation broth, as the cells were aggregated from the beginning to the end of the fermentation. Therefore, these results indicate that YZ0819 can efficiently produce succinic acid when using $NH_4OH$ as the pH controller, and the formation of aggregates can be useful for transferring the cells from a cultivation medium for various industrial applications.
An alkaliphilic bacterium, Bacillus sp. strain BK, was found to produce extracellular cellulase-free xylanolytic enzymes with xylan-binding activity. Since the pellet-bound xylanase is eluted with 2% TEA from the pellet of the culture, they contain a xylan-binding region that is stronger than the xylan-binding xylanase of the extracellular enzyme. The xylanases had a different molecular weight and xylan-binding ability. The enzyme activity of xylanase in the extracellular fraction was 6 times higher than in the pellet-bound enzyme. Among the enzymes, xylanase had the highest enzyme activity. When Bacillus sp. strain BK was grown in pH 10.5 alkaline medium containing xylan as the sole carbon source, the bacterium produced xylanase, arabinofuranosidase, acetyl esterase, and $\beta$-xylosidase with specific activities of 1.23, 0.11, 0.06, and 0.04 unit per mg of protein, respectively. However, there was no cellulase activity detected in the crude enzyme preparation. The hydrolysis of agricultural residues and kraft pulps by the xylanolytic enzymes was examined at 50$^{\circ}C$ and pH 7.0. The rate of xylan hydrolysis in com hull was higher than those of sugarcane bagasse, rice straw, com cop, rice husk, and rice bran. In contrast, the rate of xylan hydrolysis in sugarcane pulp was 2.01 and 3.52 times higher than those of eucalyptus and pine pulp, respectively. In conclusion, this enzyme can be used to hydrolyze xylan in agricultural residues and kraft pulps to breach and regenerate paper from recycled environmental resources.
Biological Treatment of Wastewater Containing Chlorinated Phenols by a Mixed Culture. Lee, Wan-Seok1, Sang-Wook Jung, Chan-Sun Park, Byung-Dae Yoon, Jang-Eok Kim\ and Hee-Mock Oh*. Environmental Bioresources Laboratory, Korea Research Institute of Biosicence and Biotechnology, Taejon, Korea, 1 Department of Agricultural Chemistry, Kyungpool< National University, Taegu, Korea - The biodegradation of chlorinated phenols in an artificial wastewater was investigated using a mixed culture. The mixed culture was composed of 8 microorganisms isolated from the soil contaminated with various chlorinated phenols. Pseudomonas sp. BM as a main constituent of a mixed culture was Gram-negative, catalase- and oxidase-positive, and rod-shaped, and did not grow at 41°C. It degraded 99% of initial 500 mg!1 of pentachlorophenol (PCP) in the minimal salts medium as a sole source of carbon and energy within 3 days. The degradation efficiency of Pseu.domon.as sp. BM was not affected by the other organic carbon and nitrogen compounds. Pseudomonas sp. BM was able to grow in a broad range of pH 5 - 8, and degrade 2,000 mg/1 PCP. In the experiment with an artificial wastewater containing chlorinated phenols, the degradation efficiency of the mixed culture was the range of 73% (2,4-dichlorophenol) -96% (2-chlorophenol) during an incubation of 7 days. In a continuous culture experiment, the degradation efficiency of mixed culture plus activated sludge was about 2 times higher than that of the control containing only activated sludge. These results indicate that it is possible to apply the mixed culture to other wastewaters containing chlorinated phenols. Key words: Biodegradation, chlorinated phenols, pentachlorophenol, Pseudomonas sp. BM
The population diversity and seasonal changes of bacterial communities in rice soils were monitored using both culture-dependent approaches and molecular methods. The rice field plot consisted of twelve subplots planted with two genetically-modified (GM) rice and two non-GM rice plants in three replicates. The DGGE analysis revealed that the bacterial community structures of the twelve subplot soils were quite similar to each other in a given month, indicating that there were no significant differences in the structure of the soil microbial populations between GM rice and non-GM rice during the experiment. However, the DGGE profiles of June soil after a sudden flooding were quite different from those of the other months. The June profiles exhibited a few intense DNA bands, compared with the others, indicating that flooding of rice field stimulated selective growth of some indigenous microorganisms. Phylogenetic analysis of l6S rDNA sequences from cultivated isolates showed that, while the isolates obtained from April soil before flooding were relatively evenly distributed among diverse genera such as Arthrobacter, Streptomyces, Terrabacter, and Bacillus/Paenibacillus, those from June soil after flooding mostly belonged to the Arthrobacter species. Phylogenetic analysis of 16S rDNA sequences obtained from the soil by cloning showed that April, August, and October had more diverse microorganisms than June. The results of this study indicated that flooding of rice fields gave a significant impact on the indigenous microbial community structure; however, the initial structure was gradually recovered over time after a sudden flooding.
Low Electromagnetic Field (EMF) intensity in the range of $1{\mu}T\;to\;10{\mu}T$(Tesla) was found to enhance the growth of CHO cells and the production of tPA in batch and perfusion cultivations. At $1{\mu}T\;intensity,\;1.3{\times}10^7$ viable cells/ml of maximum cell density and 80 mg/l of maximum tPA production were obtained in batch cultivation, compared to $2.8{\times}10^6$ viable cells/ml and 59 mg tPA/1 in unexposed case (control). A similar trend was observed in the perfusion process, where it was possible to obtain $1.2{\times}10^7$ viable cells/ml of maximum cell density and 81 mg tPA/l of maximum tPA production by more than 80 days of cultivation. However, there was not much difference between $1{\mu}T\;and\;10{\mu}T$ in perfusion cultivation, possibly due to better environmental growth conditions being maintained by continuous feeding of fresh medium into the reactor. On the contrary, both cell growth and tPA production were severely inhibited at higher than 1 mT intensity, showing no growth at 10 mT exposure. Specific growth rate was linearly correlated to specific tPA production rate at $1{\mu}T$EMF intensity, which represents a partially growth-related relationship. It was also found that a large amount of $Ca^2+$ was released at low EMF intensity, even though the cell growth was not much affected. Low EMF intensity significantly improved both cell growth and tPA production, and tPA production seemed to be more affected than the cell growth, possibly due to the changes of cell membrane characteristics. It can be concluded that the elaboration of EMF intensity less than $10{\mu}T$ could improve cell growth and tPA production, but mainly tPA secretion through batch or perfusion process in a bioreactor.
A total of 2,280 nonduplicate clinical isolates of Pseudomonas aeruginosa, obtained nationwide from Korean non-tertiary care hospitals from 2002 to 2005, were identified and their susceptibilities to aminoglycosides, antipseudomonal penicillins, carbapenems, cephalosporins, monobactams, and quinolones were studied, together with their production of ${\beta}$-lactamases. Using disk diffusion and minimum inhibitory concentration tests, it was found that 2.9% of isolates were multidrug-resistant (MDR) P. aeruginosa. An EDTA-disk synergy test, PCR amplification with specifically designed primers, and direct sequencing of the PCR products showed that the $bla_{OXA-10}$, $bla_{VIM-2}$, $bla_{OXA-2}$, $bla_{OXA-17}$, $bla_{PER-1}$, $bla_{SHV-12}$, and $bla_{IMP-1}$ genes were carried by 34.3%, 26.9%, 3.0%,3.0%, 1.5%, 1.5%, and 1.5% of 67 MDR P. aeruginosa isolates, respectively. The prevalence of MDR P. aeruginosa was three-fold higher, compared with that from the United States. More than two types of ${\beta}$-lactamase genes were carried by 10.4% of isolates. The most prevalent ${\beta}$-lactamase genes were $bla_{VIM-2}$ and $bla_{OXA-10}$. This study is the first description of MDR P. aeruginosa trom non-tertiary care hospitals in Korea and the coexistence of the $bla_{VIM-2}$, $bla_{IMP-1}$, or $bla_{PER-1} in these clinical isolates.
The fruits of sweet cherry are highly appreciated by the consumer due to their precocity and quality, such as their sweetness, color and sourness. In this study, the hot-water extract and its sequential organic solvent fractions were prepared from domestic Napoleon sweet cherry (Prunus avium L.) to investigate antimicrobial and antioxidant activity. The hot-water extract contained about 40% sugars, and the solvent fraction yields for hexane, ethylacetate (EA), butanol, and water residue were 0.01%, 3.45%, 16.30%, and 80.24%, respectively. Contents of total polyphenol and total flavonoid of the fractions were 1.24~5.24%, and 0~3.76%, respectively. Among the fractions, EA fraction showed the highest total polyphenol and total flavonoid concentrations. Evaluation of antimicrobial activity of the extract and fractions revealed that EA fraction and butanol fraction contained strong antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, and Salmonella typhimurium with minimal inhibitory concentration (MIC) of 0.5~1.0 mg/mL. But the extract and fractions tested were not active to Pseudomonas aeruginosa. In a while, only hexane fraction showed anti-Candida activity with 0.5~1.0 mg/mL of MIC. The fraction showed strong activity against different multi-antibiotics resistant strains such as C. albicans CCARM 14020. Antioxidative activity assay showed that EA fraction has a strong DPPH scavenging activity and a reducing power. The $IC_{50}s$ of vitamin E and EA fraction were 15.5 and $195.5\;{\mu}g/mL$, respectively. Our results suggest that the fruit of P. avium L. has high potentials with anti-Candida and antioxidative activity.
The global commercial cultivation of transgenic crops, including glyphosate-tolerant soybean, has increased widely in recent decades with potential impact on the environment. The bulk of previous studies showed different results on the effects of the release of transgenic plants on the soil microbial community, especially rhizosphere bacteria. In this study, comparative analyses of the bacterial communities in the rhizosphere soils and surrounding soils were performed between the glyphosate-tolerant soybean line NZL06-698 (or simply N698), containing a glyphosate-insensitive EPSPS gene, and its control cultivar Mengdou12 (or simply MD12), by a 16S ribosomal RNA gene (16S rDNA) amplicon sequencing-based Illumina MiSeq platform. No statistically significant difference was found in the overall alpha diversity of the rhizosphere bacterial communities, although the species richness and evenness of the bacteria increased in the rhizosphere of N698 compared with that of MD12. Some influence on phylogenetic diversity of the rhizosphere bacterial communities was found between N698 and MD12 by beta diversity analysis based on weighted UniFrac distance. Furthermore, the relative abundances of part rhizosphere bacterial phyla and genera, which included some nitrogen-fixing bacteria, were significantly different between N698 and MD12. Our present results indicate some impact of the glyphosate-tolerant soybean line N698 on the phylogenetic diversity of rhizosphere bacterial communities together with a significant difference in the relative abundances of part rhizosphere bacteria at different classification levels as compared with its control cultivar MD12, when a comparative analysis of surrounding soils between N698 and MD12 was used as a systematic contrast study.
In this study, two soybean genotypes, i.e., aluminum-tolerant Baxi 10 (BX10) and aluminumsensitive Bendi 2 (BD2), were used as plant materials and acidic red soil was used as growth medium. The soil layers from the inside to the outside of the root are: rhizospheric soil after washing (WRH), rhizospheric soil after brushing (BRH) and rhizospheric soil at two sides (SRH), respectively. The rhizosphere bacterial communities were analyzed by high-throughput sequencing of V4 hypervariable regions of 16S rRNA gene amplicons via Illumina MiSeq. The results of alpha diversity analysis showed that the BRH and SRH of BX10 were significantly lower in community richness than that of BD2, while the WRH exhibited no significant difference between BX10 and BD2. Among the three sampling compartments of the same soybean genotype, WRH had the lowest community richness and diversity while showing the highest coverage. Beta diversity analysis results displayed no significant difference for any compartment between the two genotypes, or among the three different sampling compartments for any same soybean genotype. However, the relative abundance of major bacterial taxa, specifically nitrogen-fixing and/or aluminum-tolerant bacteria, was significantly different in the compartments of the BRH and/or SRH at phylum and genus levels, indicating genotype-dependent variations in rhizosphere bacterial communities. Strikingly, as compared with BRH and SRH, the WRH within the same genotype (BX10 or BD2) always had an enrichment effect on rhizosphere bacteria associated with nitrogen fixation.
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