To develop multifunctional microbial inoculant, an insluble phosphate-solubilizing bacterium with antifungal activity was isolated from plant rhizospheric soil. On the basis of its morphological, cultural and physiological characteristics and Biolog analysis, this bacterium was identified as Pseudomonas fluorescens RAF15. P. fluorescens RAF15 showed antifungal activities against phytopathogenic fungi Botrytis cinerea and Rhizoctonia solani. The optimal medium composition and cultural conditions for the solubilization of insoluble phosphate by P. fluorescens RAF15 were 1.5% of glucose, 0.005% of urea, 0.3% $MgCl_2{\cdot}6H_2\;0.01%\;of\;MgSO_4{\cdot}7H_2O\;0.01%,\;of\;CaCl_2{\cdot}2H_2O$, and 0.05% of NaCl along with initial pH 7.0 at $30^{\circ}C$. The soluble phosphate production under optimum condition was 863 mg/L after 5 days of cultivation. The solubilization of insoluble phosphates was associated with a drop in the pH of the culture medium. P. fluorescens RAF15 showed resistance against different environmental stresses like $10-35^{\circ}C$ temperature, 1-4% salt concentration and pH 2-11 range. The strain produced soluble phosphate to the culture broth with the concentrations of 971-1121 mg/L against $CaHPO_4$, 791-908 mg/L against $Ca_3(PO_4){_2}$, and 844 mg/L against hydroxyapatite, respectively. However, the strain produced soluble phosphate to the culture broth with the concentrations of 15 mg/L against $FePO_4$, and 5 mg/L against $AlPO_4$, respectively.
Background : The mechanisms through which cellular activation results in intracellular mycobacterial killing is only partially understood. However, in vitro studies of human immunity to Mycobacterium tuberculosis have been largely modeled on the work reported by Crowle, which is complicated by several factors. The whole blood culture is simple and allows the simultaneous analysis of the relationship between bacterial killing and the effect of effector cells and humoral factors. In this study, we attempted to determine the extent to which M. tuberculosis is killed in a human whole blood culture and to explore the role of the host and microbial factor in this process. Methods : The PPD positive subject were compared to the umbilical cord blood and patients with tuberculosis, diabetes and lung cancer. The culture is performed using heparinized whole blood diluted with a culture medium and infected with a low number of M. avium or M. tuberculosis $H_{37}Ra$ for 4 days by rotating the culture in a $37^{\circ}C$, 5% $CO_2$ incubator. In some experiments, methlprednisolone- or pentoxifyline were used to inhibit the immune response. To assess the role of the T-cell subsets, CD4+, CD8+ T-cells or both were removed from the blood using magnetic beads. The ${\Delta}$ log killing ratio was defined using a CFU assay as the difference in the log number of viable organisms in the completed culture compared to the inoculum. Results : 1. A trend was noted toward the improved killing of mycobacteria in PPD+ subjects comparing to the umbilical cord blood but there was no specific difference in the patients with tuberculosis, diabetes and lung cancer. 2. Methylprednisolone and pentoxifyline adversely affected the killing in the PPD+ subjects umbilical cord blood and patients with tuberculosis. 3. The deletion of CD4+ or CD8+ T-lymphocytes adversely affected the killing of M. avium and M. tuberculosis $H_{37}Ra$ by PPD+ subjects. Deletion of both cell types had an additive effect, particularly in M. tuberculosis $H_{37}Ra$. 4. A significantly improved mycobacterial killing was noted after chemotherapy in patients with tuberculosis and the ${\Delta}$ logKR continuously decreased in a 3 and 4 days of whole blood culture. Conclusion : The in vitro bactericidal assay by human whole blood culture model was settled using a CFU assay. However, the host immunity to M. tuberculosis was not apparent in the human whole blood culture bactericidal assay, and patients with tuberculosis showed markedly improved bacterial killing after anti-tuberculous chemotherapy compared to before. The simplicity of a whole blood culture facilitates its inclusion in a clinical trial and it may have a potential role as a surrogate marker in a TB vaccine trial.
Journal of the Korea Organic Resources Recycling Association
/
v.9
no.1
/
pp.65-72
/
2001
Four newly isolated bacteria from soil were used to manufacture microbial inoculum to compost food waste. The bacteria, GM103, V25, V31, and V35, were identified as Bacillus licheniformis, B. subtilis, B. stearothermophilius, and B, subtilis, respectively. The bacterial strains were efficient to degrade protein and starch and also able to inhibit the growth of plant pathogenic fungus Rhizopus stronifer. The GM103 showed distinct capability in degrading starch, but grow only aerobically. The other three bacterial strains. V25, V31, and V35, could grow both aerobically as well as anaerobically, in 10%(w/v) salt, at $50^{\circ}C$, and had good viability and survival rate in soil. These characteristics of the bacterial strains are very adquate in Korean food composting containing high concentration of salt, especially at home. By mixing the 4 bacterial culture broth with molasses, beet pulp, zeolite, The bacterial inoculum for food waste composting-BIOTOP-CLEAN-was made. The performance of food waste composting by the BIOTOP-CLEAN was compared with that by control(not treated) and HS(other demestic company's inoculum product for food waste composting). The maximum temperature of the food waste during the composting with the BIOTOP-CLEAN was $50^{\circ}C$, while those of the control and HS were $30^{\circ}C$ and $35^{\circ}C$, respectively. The BIOTOP-CLEAN gave the good smell and showed dark brown color, while the control gave bad smell and HS gave less bad smell. These indicates that the food waste composting by the BIOTOP-CLEAN had been well accomplished. The culture broth of V25, V31, V35 were sparyed to the plants of tomato, chinese cabbage, raddish, red pepper every month and the spraying the culture broth to these plant significantly improved the production yield of the crops, due to the control effect of the bacterial strains against the plant pathogens.
Kim, Young-J.;Lee, Hong-C.;Park, Sung-Y.;Park, Sun-Y.;Oh, Se-Jong;Chin, Koo-B.
Food Science of Animal Resources
/
v.28
no.1
/
pp.51-58
/
2008
This study was performed to evaluate the physico-chemical properties of fermented sausages containing probiotic starter cultures (LK-30 plus, Lactobacillus plantarum 155 and 167, and Pediococcus damnosus L12) with reduced fat levels, and to determine the optimum condition for the manufacture of these products. Although low-fat fermented sausages were reduced fat content at the amount of 90% and the ripening time by 1-2 weeks, as compared to regular-fat counterpart, they became harder and had many winkles outside due to the extreme drying. In addition, fat level in fermented sausages affected the composition and shear force values. During ripening, pH, lightness and yellowness values tended to decrease, however, microbial counts of inoculated lactic acid bacteria were increased up to $10^8-10^9cfu/g$ within 3 days and remained constant thereafter. Low-fat fermented sausages had higher microbial counts than regular-fat ones. Although the inoculated probiotic starter cultures alone had the functional properties, such as cholesterol reduction, anti-high blood pressure and antimicrobial activity, they did not have distinctive characteristics in the fermented sausages. Based on these results, the low-fat fermented sausages were successfully manufactured, but a little bit increased fat level and improved functional properties in the fermented sausages would be required to have better quality as compared to regular-fat counterparts.
Kim, Min Jeong;Shim, Chang Ki;Kim, Yong Ki;Hong, Sung Jun;Park, Jong Ho;Han, Eun Jung;Kim, Jin Ho;Kim, Suk Chul
The Plant Pathology Journal
/
v.31
no.3
/
pp.259-268
/
2015
This study investigated the chemical characteristics and microbial population during incubation of four kinds of aerated compost teas based on oriental medicinal herbs compost, vermicompost, rice straw compost, and mixtures of three composts (MOVR). It aimed to determine the effects of the aerated compost tea (ACT) based on MOVR on the growth promotion of red leaf lettuce, soybean and sweet corn. Findings showed that the pH level and EC of the compost tea slightly increased based on the incubation time except for rice straw compost tea. All compost teas except for oriental medicinal herbs and rice straw compost tea contained more ${NO^-}_3-N$ than ${NH^+}_4-N$. Plate counts of bacteria and fungi were significantly higher than the initial compost in ACT. Microbial communities of all ACT were predominantly bacteria. The dominant bacterial genera were analyzed as Bacillus (63.0%), Ochrobactrum (13.0%), Spingomonas (6.0%) and uncultured bacterium (4.0%) by 16S rDNA analysis. The effect of four concentrations, 0.1%, 0.2%, 0.4% and 0.8% MOVR on the growth of red leaf lettuce, soybean and sweet corn was also studied in the greenhouse. The red leaf lettuce with 0.4% MOVR had the most effective concentration on growth parameters in foliage part. However, 0.8% MOVR significantly promoted the growth of root and shoot of both soybean and sweet corn. The soybean treated with higher MOVR concentration was more effective in increasing the root nodule formation by 7.25 times than in the lower MOVR concentrations Results indicated that ACT could be used as liquid nutrient fertilizer with active microorganisms for culture of variable crops under organic farming condition.
Perchlorate ($ClO_4^-$) is a contaminant found in surface water and soil/ground water. Microbial removal of perchlorate is the method of choice since microorganisms can reduce perchlorate into harmless end-products. Such microorganisms require an electron donor to reduce perchlorate. Conventional perchlorate-removal techniques employ heterotrophic perchlorate-reducing bacteria that use organic compounds as electron donors to reduce perchlorate. Since continuous removal of perchlorate requires a continuous supply of organic compounds, heterotrophic perchlorate removal is an expensive process. Feasibility of autotrophic perchlorate-removal using elemental sulfur granules and activated sludge was examined in this study. Granular sulfur is relatively inexpensive and activated sludge is easily available from wastewater treatment plants. Batch tests showed that activated sludge microorganisms could successfully degrade perchlorate in the presence of granular sulfur as an electron donor. Perchlorate biodegradation was confirmed by molar yield of $Cl^-$ as the perchlorate was degraded. Scanning electron microscope revealed that rod-shaped microorganisms on the surface of sulfur particles were used for the autotrophic perchlorate-removal, suggesting that sulfur particles could serve as supporting media for the formation of biofilm as well. DGGE analyses revealed that microbial profile of the inoculum (activated sludge) was different from that of the biofilm sample obtained from enrichment culture that used sulfur particles for $ClO_4^-$-degradation.
Kim, Mi-Sun;Hong, Young-Ah;Yeo, Soo-Hwan;Baek, Seong-Yeol;Yun, Hye-Ju;Rhee, Chang-Ho;Kim, Kwan-Pil;Park, Heui-Dong
Food Science and Preservation
/
v.20
no.6
/
pp.886-893
/
2013
Several indigenous sulfite-resistant yeasts were isolated at the microbial succession stage of yeast flora during spontaneous fermentation of Campbell Early grapes using a YPD plate that contained 200 mg/L or 500 mg/L potassium metabisulfite. When they were applied to the wine fermentation using the Campbell Early grape and apple juices, strains S13 and D8 showed strong alcohol fermentation and good flavor production. They were identified as Saccharomyces cerevisiae in the phylogenetic analysis based on their ITS 1-5.8S-ITS II DNA sequences. The two yeast strains grew to a high cell density in the YPD media supplemented with 40%(w/v) glucose. They also grew rapidly in the YPD media at $40^{\circ}C$. While strain S13 showed some differences in cell density at the two temperatures, no marked difference was observed during the culture of strain D8. The strains grew relatively well at pH 5.0 and 9.0 compared with pH 7.0, which was the optimum pH for their growth. Especially, strain S13 cultivated in the YPD media at pH 9.0 grew to 93% of the growth of strain D8, which was obtained at pH 7.0.
Jeong, Yong Dae;Lee, Jung Jae;Seol, Kuk-Hwan;Kim, Doo Wan;Min, Ye Jin;Yu, Dong Jo;Cho, Kyu Ho;Kim, Young Hwa
Korean Journal of Agricultural Science
/
v.44
no.4
/
pp.558-565
/
2017
This study was conducted to determine the effect of inoculation of microorganism isolated from pig feces on nutrient contents of fermented hulless barley. The microbial flora in feces of a total of four crossbred piglets ($Landrace{\times}Yorkshire{\times}Duroc$) was analyzed by 16s rRNA sequencing. The most abundant strain was then selected for fermentation of hulless barley. Lactobacillus plantarum (L. plantarum) was dominant (64.56%) in intestinal microbial flora in the pig feces. The selected candidate strain showed significantly higher survival rate at pH 7 than at pH 2.5 and 3.0 (p < 0.05). Incubated culture containing the candidate strain showed an increased growth rate with lower pH levels after 7.5 h incubation compared to initial incubation period (p < 0.05). When compared with commercial multiple probiotics which were used as control, the selected strain showed faster growth rate at 5 h post-incubation (p < 0.05). During the fermentation period, neither inoculated nor non-inoculated control hulless barley showed any change in pH value. Crude fat, fiber and ash contents were lower (p < 0.05) in hulless barley inoculated by the selected strain compared to control. However, moisture, energy, NDF and ADF were not affected by the inoculation of strain or fermentation period. Lactic acid was increased and acetic acid was decreased in the hulless barley inoculated with the selected strain during the fermentation period (p < 0.05). Taken together, our results suggest that L. plantarum derived from the pigs could be utilized as a new microorganism for manufacturing fermented feed stuffs.
Microbial desulfurization characterlstics of a bituminous coal have been determined by using Thiobacillus ferrooxidans. The effects of process variables (such as coal pulp density, particle size and addition of surfactants) on pyrite removal have been investigated in both shake and airlift-bioreactor culture experiments. In shake experiments, pyrite could be removed over 78% for pulp densifies below 20% (w/v) and removed below 40% for pulp densities over 30% (w/v) in 8 days. Pyrite removal decreased with increasing pulp densities, and it also decreased sharply with increasing particle sizes. In airlift bioreactor experiments, pyrite at 50% (w/v) pulp density could be removed about 50%. Its value is much higher than 15% at the same pulp density in a shake experiment. With addition of surfactants, pyrite removal was enhanced in shake experiments significantly, whereas it was slightly decreased in an airlift bioreactor experiment.
Maillard reaction products (MRPs) added into a culture and the resultant bacterial growth were investigated using response surface methodology. The coefficients of determination $(R^{2})$ of response surface regression equations for bacteria were 0.9544 and 0.9578 in Bacillus subtilis and Bacillus natto, respectively. The MRPs produced at higher reaction temperature and for longer reaction time showed greater antimicrobial effect for Bacillus subtilis. Especially, the MRPs produced at temperature above $150^{\circ}C$ for 8 to 12 hrs showed the strongest antimicrobial effect. The MRPs produced at lower reaction temperature and for shorter reaction time showed greater microbial growth effect for Bacillus natto, but those produced at the reaction temperature higher than $160^{\circ}C$ showed the greatest antimicrobial effect. In the ridge analysis, the growth of Bacillus subtilis was the most significantly inhibited in the presence of MRPs prepared at $159.10^{\circ}C$ and pH 12.21 for 9.67 hrs, and the growth of Bacillus natto was the most significantly inhibited in the presence of MRPs prepared at $169.94^{\circ}C$ and pH 9.66 for 9.22 hrs.
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